sop - quy trình vi sinh vật
TRANSCRIPT
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B Y T
S: 26 /Q-BYT
CNG HA X HI CH NGHA VIT NAM
c lp - T do - Hnh phc
H Ni, ngy 03 thng 01 nm 2013
QUYT NH
V vic ban hnh ti liu
Hng dn quy trnh k thut chuyn ngnh Vi sinh Y hc
B TRNG B Y T
Cn c Lut khm bnh, cha bnh nm 2009;
Cn c Ngh nh s 63/2012/N-CP ngy 31/8/2012 ca Chnh Ph quy
nh chc nng, nhim v, quyn hn v c cu t chc ca B Y t;
Xt Bin bn hp ca Hi ng nghim thu Hng dn Quy trnh k
thut khm bnh, cha bnh chuyn ngnh Vi sinh Y hc ca B Y t;
Theo ngh ca Cc trng Cc Qun l Khm, cha bnh,
QUYT NH:
iu 1. Ban hnh km theo Quyt nh ny ti liu Hng dn quy trnh
k thut chuyn ngnh Vi sinh Y hc, gm 231 quy trnh k thut.
iu 2. Ti liu Hng dn quy trnh k thut chuyn ngnh Vi sinh Y
hc ban hnh km theo Quyt nh ny c p dng ti cc c s khm bnh,
cha bnh.
Cn c vo ti liu hng dn ny v iu kin c th ca n v, Gim
c c s khm bnh, cha bnh xy dng v ban hnh ti liu Hng dn Quy
trnh k thut Vi sinh Y hc ph hp thc hin ti n v.
iu 3. Quyt nh ny c hiu lc k t ngy k ban hnh.
iu 4. Cc ng, b: Chnh Vn phng B, Chnh Thanh tra B, Cc
trng Cc Qun l Khm, cha bnh, Cc trng v V trng cc Cc, V
thuc B Y t, Gim c cc bnh vin, vin c ging bnh trc thuc B Y
t, Gim c S Y t cc tnh, thnh ph trc thuc trung ng, Th trng Y
t cc B, Ngnh v Th trng cc n v c lin quan chu trch nhim thi
hnh Quyt nh ny./.
Ni nhn: - Nh iu 4;
- B trng B Y t ( b/c);
- Cc Th trng BYT;
- Bo him X hi Vit Nam ( phi hp);
- Cng thng tin in t BYT;
- Website Cc KCB;
- Lu VT, KCB.
KT. B TRNG
TH TRNG
k
Nguyn Th Xuyn
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B Y T CNG HA X HI CH NGHA VIT NAM
c lp - T do- Hnh Phc
DANH SCH CC QUY TRNH K THUT
CHUYN NGNH VI SINH HC
(Ban hnh km theo Quyt nh s: 26/Q-BYT ngy 03 thng 01 nm 2014
ca B trng B Y t)
TT TN QUY TRNH K THUT
Danh mc xt nghim vi khun
A. Vi khun chung
1 Vi khun nhum soi
2 Vi khun test nhanh
3 Vi khun nui cy v nh danh phng php thng thng
4 Vi khun nui cy v nh danh h thng t ng
5 Vi khun nui cy, nh danh v khng thuc h thng t ng
6 Vi khun khng thuc nh tnh
7 Vi khun khng thuc h thng t ng
8 Vi khun khng thuc nh lng (MIC) (cho 1 loi khng sinh)
9 Vi khun khng sinh phi hp
10 Vi khun k kh nui cy v nh danh
11 Vi khun khng nh
12 Vi khun nh danh PCR
13 Vi khun nh danh gii trnh t gene
14 Vi khun khng thuc PCR
15 Vi khun khng thuc gii trnh t gene
16 Vi h ng rut
B. Mycobacteria
17 AFB trc tip nhum Ziehl-Neelsen
18 AFB trc tip nhum hunh quang
19 Mycobacterium tuberculosis nui cy mi trng lng
20 Mycobacterium tuberculosis nui cy mi trng c
21 Mycobacterium tuberculosis Mantoux
22 Mycobacterium tuberculosis khng thuc hng 1 mi trng c
23 Mycobacterium tuberculosis khng thuc hng 1 mi trng lng
24 Mycobacterium tuberculosis khng thuc hng 2 mi trng c
25 Mycobacterium tuberculosis khng thuc hng 2 mi trng lng
26 Mycobacterium tuberculosis khng thuc PZA mi trng lng
27 Mycobacterium tuberculosis pyrazinamidase
28 Mycobacterium tuberculosis nh danh v khng RMP Xpert
29 Mycobacterium tuberculosis a khng LPA
30 Mycobacterium tuberculosis siu khng LPA
31 Mycobacterium tuberculosis PCR h thng t ng
32 Mycobacterium tuberculosis Real-time PCR
33 Mycobacterium tuberculosis spoligotyping
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34 Mycobacterium tuberculosis RFLP typing
35 NTM (Non tuberculosis mycobacteria) nui cy mi trng lng
36 NTM (Non tuberculosis mycobacteria) nui cy mi trng c
37 NTM (Non tuberculosis mycobacteria) nh danh LPA
38 Mycobacterium leprae nhum soi
39 Mycobacterium leprae PCR
40 Mycobacterium leprae mnh sinh thit
C. Vibrio cholerae
41 Vibrio cholerae nhum soi
42 Vibrio cholerae nhum hunh quang
43 Vibrio cholerae nui cy, nh danh v khng thuc
44 Vibrio cholerae PCR
45 Vibrio cholerae gii trnh t gene
D. Neisseria gonorrhoeae
46 Neisseria gonorrhoeae nhum soi
47 Neisseria gonorrhoeae nui cy, nh danh v khng thuc
48 Neisseria gonorrhoeae PCR
49 Neisseria gonorrhoeae Real-time PCR
E. Neisseria meningitidis
50 Neisseria meningitidis nhum soi
51 Neisseria meningitidis nui cy, nh danh v khng thuc
52 Neisseria meningitidis PCR
53 Neisseria meningitidis Real-time PCR
F. Cc vi khun khc
54 Chlamydia test nhanh
55 Chlamydia nhum hunh quang
56 Chlamydia Ab min dch bn t ng
57 Chlamydia PCR
58 Chlamydia Real-time PCR
59 Chlamydia Real-time PCR h thng t ng
60 Clostridium nui cy, nh danh
61 Clostridium difficile min dch bn t ng
62 Clostridium difficile PCR
63 Leptospira test nhanh
64 Mycoplasma pneumoniae Real-time PCR
65 Mycoplasma hominis nui cy, nh danh v khng thuc
66 Rickettsia Ab min dch bn t ng
67 Salmonella Widal
68 Streptococcus pyogenes ASO
69 Treponema pallidum soi ti
70 Treponema pallidum nhum soi
71 Treponema pallidum RPR nh tnh v nh lng
72 Treponema pallidum TPHA nh tnh v nh lng
73 Ureaplasma urealyticum nui cy, nh danh v khng thuc
Danh mc xt nghim virus
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A. Virus chung
74 Virus test nhanh
75 Virus Ag min dch bn t ng
76 Virus Ab min dch bn t ng
B. Hepatitis virus
77 HBsAg test nhanh
78 HBsAg min dch bn t ng
79 HBsAg min dch t ng
80 HBsAg khng nh
81 HBsAg nh lng
82 HBsAb min dch bn t ng
83 HBsAb nh lng
84 HBc IgM min dch bn t ng
85 HBc IgM min dch t ng
86 HBc total min dch bn t ng
87 HBc total min dch t ng
88 HBeAg min dch bn t ng
89 HBeAg min dch t ng
90 HBeAb min dch bn t ng
91 HBeAb min dch t ng
92 HBV o ti lng Real-time PCR
93 HBV o ti lng h thng t ng
94 HBV genotype PCR
95 HBV genotype Real-time PCR
96 HBV genotype gii trnh t gene
97 HBV khng thuc Real-time PCR (cho 1 loi thuc)
98 HBV khng thuc gii trnh t gene
99 HCV Ab min dch bn t ng
100 HCV Ab min dch t ng
101 HCV Ag/Ab min dch bn t ng
102 HCV Core Ag min dch t ng
103 HCV PCR
104 HCV o ti lng Real-time PCR
105 HCV o ti lng h thng t ng
106 HCV genotype Real-time PCR
107 HCV genotype gii trnh t gene
108 HAV IgM min dch bn t ng
109 HAV IgM min dch t ng
110 HAV total min dch bn t ng
111 HAV total min dch t ng
112 HDV Ag min dch bn t ng
113 HDV IgM min dch bn t ng
114 HDV Ab min dch bn t ng
115 HEV IgM test nhanh
116 HEV IgM min dch bn t ng
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C. HIV
117 HIV Ab test nhanh
118 HIV Ag/Ab test nhanh
119 HIV Ab min dch bn t ng
120 HIV Ag/Ab min dch bn t ng
121 HIV Ag/Ab min dch t ng
122 HIV khng nh
123 HIV o ti lng Real-time PCR
124 HIV o ti lng h thng t ng
125 HIV genotype gii trnh t gene
126 HIV khng thuc gii trnh t gene
D. Dengue virus
127 Dengue virus NS1Ag test nhanh
128 Dengue virus NS1Ag/IgM/IgG test nhanh
129 Dengue virus IgM/IgG test nhanh
130 Dengue virus IgM min dch bn t ng
131 Dengue virus IgG min dch bn t ng
132 Dengue virus PCR
133 Dengue virus serotype PCR
E. Herpesviridae
134 CMV IgM min dch bn t ng
135 CMV IgM min dch t ng
136 CMV IgG min dch bn t ng
137 CMV IgG min dch t ng
138 CMV PCR
139 CMV Real-time PCR
140 CMV o ti lng h thng t ng
141 CMV Avidity
142 HSV 1+2 IgM min dch bn t ng
143 HSV 1+2 IgG min dch bn t ng
144 HSV Real-time PCR
145 VZV Real-time PCR
146 EBV-VCA IgM min dch bn t ng
147 EBV-VCA IgG min dch bn t ng
148 EBV EA-D IgG min dch bn t ng
149 EBV EB-NA IgG min dch bn t ng
150 EBV PCR
151 EBV Real-time PCR
E. Enterovirus
152 EV71 PCR
153 EV71 Real-time PCR
154 EV71 genotype gii trnh t gene
155 Enterovirus PCR
156 Enterovirus genotype gii trnh t gene
F. Cc virus khc
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157 Adenovirus Real-time PCR
158 BK/JC virus PCR
159 HPV PCR
160 HPV Real-time PCR
161 HPV genotype Real-time PCR
162 HPV genotype PCR h thng t ng
163 HPV genotype gii trnh t gene
164 Influenza virus A, B test nhanh
165 Influenza virus A, B Real-time PCR
166 Influenza virus A, B gii trnh t gene
167 JEV IgM min dch bn t ng
168 Measles virus Ab min dch bn t ng
169 Rotavirus test nhanh
170 RSV Ab min dch bn t ng
171 RSV Real-time PCR
172 Rubella virus IgM min dch bn t ng
173 Rubella virus IgM min dch t ng
174 Rubella virus IgG min dch bn t ng
175 Rubella virus IgG min dch t ng
176 Rubella virus PCR
177 Rubella virus gii trnh t gene
Danh mc xt nghim KST
A. K sinh trng trong phn
178 Hng cu, bch cu trong phn soi ti
179 Hng cu trong phn test nhanh
180 n bo ng rut soi ti
181 n bo ng rut nhum soi
182 Trng giun, sn soi ti
183 Trng giun soi tp trung
184 Strongyloides stercoralis (giun ln) u trng soi ti
B. K sinh trng trong mu
185 Angiostrogylus cantonensis (Giun trn chut) Ab min dch bn t ng
186 Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t ng
187 Cysticercus cellulosae (Sn ln) Ab min dch bn t ng
188 Entamoeba histolytica (Amip) Ab min dch bn t ng
189 Fasciola (Sn l gan ln) Ab min dch bn t ng
190 Filaria (Giun ch) u trng trong mu nhum soi
191 Gnathostoma Ab min dch bn t ng
192 Plasmodium (K sinh trng st rt) nhum soi nh tnh
193 Plasmodium (K sinh trng st rt) nhum soi nh lng
194 Plasmodium (K sinh trng st rt) Ag test nhanh
195 Strongyloides stercoralis (Giun ln) Ab min dch bn t ng
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196 Toxocara (Giun a ch, mo) Ab min dch bn t ng
197 Toxoplasma IgM min dch t ng
198 Toxoplasma IgG min dch t ng
199 Toxoplasma Avidity
C. K sinh trng ngoi da
200 Demodex soi ti
201 Demodex nhum soi
202 Phthirus pubis (Rn mu) soi ti
203 Phthirus pubis (Rn mu) nhum soi
204 Sarcoptes scabies hominis(Gh) soi ti
205 Sarcoptes scabies hominis(Gh) nhum soi
D. K sinh trng trong cc bnh phm khc
206 Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit
207 Gnathostoma u trng soi mnh sinh thit
208 Pneumocystis jirovecii nhum soi
209 Taenia (Sn dy) soi ti nh danh
210 Toxocara (Giun a ch, mo) soi mnh sinh thit
211 Trichinella spiralis (Giun xon) soi mnh sinh thit
212 Trichomonas vaginalis soi ti
213 Trichomonas vaginalis nhum soi
Danh mc xt nghim Vi nm
214 Vi nm soi ti
215 Vi nm test nhanh
216 Vi nm nhum soi
217 Vi nm nui cy v nh danh phng php thng thng
218 Vi nm nui cy v nh danh h thng t ng
219 Vi nm nui cy, nh danh v khng thuc h thng t ng
220 Vi nm khng nh
221 Vi nm khng thuc nh lng (MIC) (cho 1 loi khng sinh)
222 Vi nm PCR
223 Vi nm gii trnh t gene
Danh mc xt nghim nh gi nhim khun bnh vin
224 Vi sinh vt cy kim tra khng kh
225 Vi sinh vt cy kim tra bn tay
226 Vi sinh vt cy kim tra dng c tit trng
227 Vi sinh vt cy kim tra b mt
228 Vi sinh vt cy kim tra nc sinh hot
229 Vi sinh vt cy kim tra nc thi
230 Vi khun khng thuc - Pht hin ngi mang
231 Vi khun gy nhim trng bnh vin - Pht hin ngun nhim
(Tng s 231 quy trnh k thut) KT. B TRNG
TH TRNG
Nguyn Th Xuyn
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MC LC
XT NGHIM VI KHUN
1. Vi khun nhum soi .......................................................................................... 1
2. Vi khun test nhanh ........................................................................................... 4
3. Vi khun nui cy v nh danh phng php thng thng ........................... 7
4. Vi khun nui cy, nh danh h thng t ng ............................................. 10
5. Vi khun nui cy, nh danh v khng thuc h thng t ng ................... 14
6. Vi khun khng thuc nh tnh ...................................................................... 18
7. Vi khun khng thuc h thng t ng ......................................................... 21
8. Vi khun khng thuc nh lng MIC (cho 1 loi khng sinh) .................... 24
9. Vi khun khng sinh phi hp ........................................................................ 28
10. Vi khun k kh nui cy v nh danh ......................................................... 31
11. Vi khun khng nh ..................................................................................... 34
13. Vi khun nh danh gii trnh t gene........................................................... 40
14. Vi khun khng thuc PCR ........................................................................... 44
15. Vi khun khng thuc gii trnh t gene ....................................................... 47
16. Vi h ng rut ........................................................................................... 51
17. AFB trc tip nhum Ziehl-Neelsen ............................................................. 54
18. AFB trc tip nhum hunh quang ............................................................... 58
19. Mycobacterium tuberculosis nui cy mi trng lng ............................... 61
20. Mycobacterium tuberculosis nui cy mi trng c ................................. 66
21. Mycobacterium tuberculosis Mantoux ......................................................... 71
22. Mycobacterium tuberculosis khng thuc hng 1 mi trng c ............... 74
23. Mycobacterium tuberculosis khng thuc hng 1 mi trng lng ............. 79
24. Mycobacterium tuberculosis khng thuc hng 2 mi trng c ............... 83
25. Mycobacterium tuberculosis khng thuc hng 2 mi trng lng ............. 88
26. Mycobacterium tuberculosis khng thuc PZA mi trng lng ................ 92
27. Mycobacterium tuberculosis pyrazinamidase ............................................... 96
28. Mycobacterium tuberculosis nh danh v khng RMP XPERT ................. 99
29. Mycobacterium tuberculosis a khng LPA ............................................... 102
30. Mycobacterium tuberculosis siu khng LPA ............................................ 107
31. Mycobacterium tuberculosis PCR h thng t ng .................................. 113
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32. Mycobacterium tuberculosis Real-time PCR ............................................. 116
33. Mycobacterium tuberculosis Spoligotyping ............................................... 120
34. Mycobacterium tuberculosis RFLP typing ................................................. 125
35. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng lng .......... 131
36. NTM (Non Tuberculosis Mycobacteria) nui cy mi trng c ............ 136
37. NTM (Non Tuberculosis Mycobacteria) LPA ............................................ 141
38. Mycobacterium leprae nhum soi .............................................................. 145
39. Mycobacterium leprae PCR ....................................................................... 148
40. Mycobacterium leprae mnh sinh thit ...................................................... 152
41. Vibrio cholerae nhum soi ......................................................................... 154
42. Vibrio cholerae nhum hunh quang ......................................................... 157
43. Vibrio cholerae nui cy, nh danh v khng thuc ................................. 160
44. Vibrio cholerae PCR ................................................................................... 163
45. Vibrio cholerae gii trnh t gene ............................................................... 166
46. Neisseria gonorrhoeae nhum soi .............................................................. 169
47. Neisseria gonorrhoeae nui cy, nh danh v khng thuc ..................... 172
48. Neisseria gonorrhoeae PCR ....................................................................... 175
49. Neisseria gonorrhoeae Real-time PCR ...................................................... 178
50. Neisseria meningitidis nhum soi ............................................................... 182
51. Neisseria meningitidis nui cy, nh danh v khng thuc ...................... 185
52. Neisseria menigititdis PCR ......................................................................... 188
54. Chlamydia test nhanh .................................................................................. 195
55. Chlamydia nhum hunh quang ................................................................. 198
56. Chlamydia Ab min dch bn t ng ........................................................ 201
57. Chlamydia PCR .......................................................................................... 205
58. Chlamydia Real-time PCR.......................................................................... 209
59. Chlamydia Real-time PCR h thng t ng ............................................. 213
60. Clostridium nui cy, nh danh ................................................................. 216
61. Clostridium difficile min dch bn t ng ............................................... 219
62. Clostridium difficile PCR............................................................................ 222
63. Leptospira test nhanh .................................................................................. 225
64. Mycoplasma pneumoniae Real-time PCR .................................................. 227
65. Mycoplasma hominis nui cy, nh danh v khng thuc ........................... 231
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66. Ricketsia Ab min dch bn t ng ........................................................... 234
67. Salmonella Widal ........................................................................................ 237
68. Streptococcus pyogenes ASO ..................................................................... 240
69. Treponema pallidum soi ti ...................................................................... 243
70. Treponema pallidum nhum soi ................................................................. 245
71. Treponema pallidum RPR nh tnh v nh lng .................................... 248
72. Treponema pallidum TPHA nh tnh v nh lng ................................. 251
73. Ureaplasma urealyticum nui cy, nh danh v khng thuc ...................... 255
XT NGHIM VIRUS
74. Virus test nhanh ........................................................................................... 258
75. Virus Ag min dch t ng ........................................................................ 260
76. Virus Ab min dch t ng ........................................................................ 264
77. HBsAg test nhanh ....................................................................................... 268
78. HBsAg min dch bn t ng .................................................................... 271
79. HBsAg min dch t ng........................................................................... 275
80. HBsAg khng nh ...................................................................................... 279
81. HBsAg nh lng ...................................................................................... 283
82. HBsAb min dch bn t ng .................................................................... 287
83. HBsAb nh lng ...................................................................................... 291
84. HBc IgM min dch bn t ng ................................................................ 295
85. HBc IgM min dch t ng ....................................................................... 299
86. HBc total min dch bn t ng ................................................................ 303
87. HBc total min dch t ng ....................................................................... 307
88. HBeAg min dch bn t ng ................................................................... 311
89. HBeAg min dch t ng .......................................................................... 315
90. HBeAb min dch bn t ng ................................................................... 318
91. HBeAb min dch t ng .......................................................................... 322
92. HBV o ti lng Real-time PCR .............................................................. 325
93. HBV o ti lng h thng t ng ............................................................ 329
94. HBV genotype PCR .................................................................................... 333
95. HBV genotype Real-time PCR ................................................................... 336
96. HBV genotype gii trnh t gene ................................................................ 340
97. HBV khng thuc Real-time PCR (Cho 1 loi thuc) ................................ 343
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98. HBV khng thuc gii trnh t gene ........................................................... 346
99. HCV Ab min dch bn t ng ................................................................. 349
100. HCV Ab min dch t ng ...................................................................... 353
101. HCV Ag/Ab min dch bn t ng ......................................................... 357
102. HCV Core Ag min dch t ng ............................................................. 361
103. HCV PCR .................................................................................................. 365
104. HCV o ti lng Real-time PCR ............................................................ 368
105. HCV o ti lng h thng t ng ......................................................... 372
106. HCV genotype Real-time PCR ................................................................. 375
107. HCV genotype gii trnh t gene .............................................................. 379
108. HAV IgM min dch bn t ng ............................................................. 382
109. HAV IgM min dch t ng .................................................................... 386
110. HAV total min dch bn t ng ............................................................. 390
111. HAV total min dch t ng ................................................................... 394
112. HDV Ag min dch bn t ng ............................................................... 398
113. HDV IgM min dch bn t ng ............................................................. 402
114. HDV Ab min dch bn t ng ............................................................... 406
115. HEV IgM test nhanh ................................................................................. 409
116. HEV IgM min dch bn t ng ............................................................. 412
117. HIV Ab test nhanh .................................................................................... 415
118. HIV Ag/Ab test nhanh .............................................................................. 418
120. HIV Ag/Ab min dch bn t ng .......................................................... 425
121. HIV Ag/Ab min dch t ng ................................................................. 429
122. HIV khng nh ........................................................................................ 433
123. HIV o ti lng Real-time PCR.............................................................. 437
124. HIV o ti lng h thng t ng ........................................................... 440
125. HIV genotype gii trnh t gene ............................................................... 444
128. Dengue virus NS1Ag/IgM/IgG test nhanh ............................................... 453
129. Dengue virus IgM/IgG test nhanh ............................................................ 456
130. Dengue virus IgM min dch bn t ng ................................................ 459
131. Dengue virus IgG min dch bn t ng ................................................. 463
132. Dengue virus PCR .................................................................................... 467
133. Dengue virus serotype PCR ...................................................................... 470
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134. CMV IgM min dch bn t ng ............................................................. 473
135. CMV IgM min dch t ng ................................................................... 477
136. CMV IgG min dch bn t ng ............................................................. 481
137. CMV IgG min dch t ng .................................................................... 485
138. CMV PCR ................................................................................................. 489
139. CMV Real-time PCR ................................................................................ 492
140. CMV o ti lng h thng t ng ......................................................... 495
141. CMV Avidity ............................................................................................ 498
142. HSV 1 + 2 IgM min dch bn t ng .................................................... 502
143. HSV 1+ 2 IgG min dch bn t ng ...................................................... 506
144. HSV Real-time PCR ................................................................................. 510
145. VZV Real-time PCR ................................................................................. 514
146. EBV-VCA IgM min dch bn t ng .................................................... 518
147. EBV-VCA IgG min dch bn t ng ..................................................... 522
148. EBV EA-D IgG min dch bn t ng .................................................... 526
149. EBV EB-NA IgG min dch bn t ng ................................................. 530
150. EBV PCR .................................................................................................. 534
151. EBV Real-time PCR ................................................................................. 537
152. EV71 PCR ................................................................................................. 541
155. Enterovirus PCR........................................................................................ 551
156. Entrovirus genotype gii trnh t gene ...................................................... 554
157. Adenovirus Real-time PCR ....................................................................... 557
158. BK/JC virus PCR ...................................................................................... 561
159. HPV PCR .................................................................................................. 565
160. HPV Real-time PCR ................................................................................. 568
161. HPV Genotype Real-time PCR ................................................................. 572
162. HPV genotype PCR h thng t ng ...................................................... 575
163. HPV genotype gii trnh t gene ............................................................... 579
164. Influenza virus A, B test nhanh ................................................................. 582
165. Influenza virus A, B Real-time PCR ......................................................... 584
166. Influenza virus A, B gii trnh t gene ..................................................... 587
167. JEV IgM min dch bn t ng ............................................................... 590
168. Measle virus Ab min dch bn t ng ................................................... 593
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169. Rotavirus test nhanh .................................................................................. 596
170. RSV Ab min dch bn t ng ................................................................ 599
171. RSV Real-time PCR ................................................................................. 602
172. Rubella virus IgM min dch bn t ng ................................................ 605
173. Rubella virus IgM min dch t ng ....................................................... 609
174. Rubella virus IgG min dch bn t ng ................................................. 613
175. Rubella virus IgG min dch t ng ....................................................... 617
176. Rubella virus PCR .................................................................................... 621
177. Rubella virus gii trnh t gene................................................................. 624
XT NGHIM K SINH TRNG - VI NM
178. Hng cu, bch cu trong phn soi ti ................................................... 627
179. Hng cu trong phn test nhanh ............................................................... 630
180. n bo ng rut soi ti ..................................................................... 633
181. n bo ng rut nhum soi ................................................................ 636
182. Trng giun, sn soi ti ............................................................................ 639
183. Trng giun soi tp trung ........................................................................... 642
184. Strongyloides stercoralis (Giun ln) u trng soi ti ........................... 645
185. Angiostrongylus cantonensis (Giun trn chut) Ab min dch bn t
ng .......................................................................................................... 648
186. Clonorchis/Opisthorchis (Sn l gan nh) Ab min dch bn t
ng .......................................................................................................... 651
187. Cysticercus cellulosae (Sn ln) Ab min dch bn t ng.................... 654
188. Entamoeba histolytica (Amip) Ab min dch bn t ng ....................... 657
189. Fasciola (Sn l gan ln) Ab min dch bn t ng ............................... 660
190. Filaria (Giun ch) u trng trong mu nhum soi .................................... 663
191. Gnathostoma(Giun u gai) Ab Min dch bn t ng.......................... 666
192. Plasmodium (K sinh trng st rt) nhum soi nh tnh ........................ 669
193. Plasmodium (K sinh trng st rt) nhum soi nh lng ..................... 672
194. Plasmodium (K sinh trng st rt) Ag test nhanh .................................. 675
195. Strongyloides stercoralis (Giun ln) Ab min dch bn t ng ........... 677
196. Toxocara (Giun a ch mo) Ab min dch bn t ng ....................... 680
197. Toxoplasma IgM min dch t ng ......................................................... 683
198. Toxoplasma IgG min dch t ng ......................................................... 687
199. Toxoplasma Avidity min dch t ng ................................................... 691
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200. Demodex soi ti ...................................................................................... 695
201. Demodex nhum soi .................................................................................. 698
202. Phthirus pubis (Rn mu) soi ti .............................................................. 701
203. Phthirus pubis (Rn mu) nhum soi ......................................................... 703
204. Sarcoptes scabies hominis (Gh) soi ti ................................................. 706
205. Sarcoptes scabies hominis (Gh) nhum soi ............................................ 709
206. Cysticercus cellulosae (Sn ln) u trng soi mnh sinh thit ................. 712
207. Gnathostoma u trng soi mnh sinh thit ............................................... 715
208. Taenia (Sn dy) soi ti nh danh ........................................................ 718
209. Toxocara (Giun a ch, mo) soi mnh sinh thit ................................ 721
210. Trichinella spiralis (Giun xon) soi mnh sinh thit ................................ 724
211. Trichomonas vaginalis soi toi ................................................................ 727
212. Trichomonas vaginalis nhum soi ............................................................ 730
213. Pneumocystis jirovecii nhum soi............................................................. 733
214. Vi nm soi ti .......................................................................................... 736
215. Vi nm test nhanh...................................................................................... 739
216. Vi nm nhum soi ..................................................................................... 742
217. Vi nm nui cy v nh danh bng phng php thng thng ............. 745
218. Vi nm nui cy v nh danh h thng t ng ...................................... 748
219. Vi nm nui cy, nh danh v khng thuc h thng t ng ................ 751
220. Vi nm khng nh (tham chiu)............................................................... 754
221. Vi nm khng thuc nh lng (MIC) (Cho 1 loi khng sinh) ............. 757
222. Vi nm PCR .............................................................................................. 760
223. Vi nm gii trnh t gene .......................................................................... 763
XT NGHIM NH GI NHIM KHUN BNH VIN
224. Vi sinh vt cy kim tra khng kh ........................................................... 766
225. Vi sinh vt cy kim tra bn tay ................................................................ 770
226. Vi sinh vt cy kim tra dng c tit trng .......................................... 773
227. Vi sinh vt cy kim tra b mt................................................................. 776
228. Vi sinh vt cy kim tra nc sinh hot .................................................... 779
229. Vi sinh vt cy kim tra nc thi ............................................................ 783
230. Vi khun khng thuc - Pht hin ngi mang ........................................ 787
231. Vi khun gy nhim trng bnh vin - Pht hin ngun nhim ............... 841
-
CC T VIT TT
BCN: Bch cu a nhn
CLIA: Chemiluminescence immunoassay
CLSI: Clinical and laboratory standards institute
CO: Cut off
DNA: Deoxyribonucleic acid
ELISA: Enzyme-linked immunosorbant assay
EQAS: External quality assessment scheme
HBV: Hepatitis B virus
HCV: Hepatitis C virus
HIV: Human immunodeficiency virus
HPC: High positive control
HT: Huyt thanh
I: Intermediate
IU: International unit
KKT: Khng khng th
KN-KT: Khng nguyn - Khng th
KST: K sinh trng
LPC: Low positive control
NC: Negative control
OD: Optical density
PC: Positive control
PCR: Polymerase chain reaction
QC: Quality control
R: Resistant
RNA: Ribonucleic acid
RT PCR: Reverse transcription polymerase chain reaction
S: Susceptible
VD: V d
VK: Vi khun
VSV: Vi sinh vt
-
1
1. Vi khun nhum soi
I. MC CH V NGUYN L
1. Mc ch
Nhn nh s b hnh nh vi khun v cc hnh nh t bo (nu c) trc
tip t bnh phm.
2. Nguyn l
nh gi hnh th, kch thc, tnh cht bt mu, cch sp xp ca vi
khun v cc hnh nh t bo (nu c) bng k thut nhum v soi di knh
hin vi quang hc.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- My ly tm (cn cho mt s loi bnh phm)
- T an ton sinh hc cp 2
- Knh hin vi quang hc
- Dng c sy lam (nu c)
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Que cy Ci 1,000
3 Lam knh Ci 2,000
4 Du soi knh ml 1,000
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
7 Thuc nhum fuchsin ml 5,000
8 Thuc nhum tm gentian ml 5,000
9 Cn ty 95% ml 10,000
-
2
10 Lugol ml 5,000
11 Thuc nhum xanh methylen ml 5,000
12 Bng Kg 0,001
13 Cn 90 (v sinh dng c) ml 10,000
14 n cn Ci 0,0001
15 Panh Ci 0,0001
16 Khay ng bnh phm Ci 0,0001
17 Hp vn chuyn bnh phm Ci 0,0001
18 M Ci 0,020
19 Khu trang Ci 0,020
20 Gng tay i 3,000
21 Gng tay x l dng c i 0,020
22 Qun o bo h B 0,001
23 Acid ngm lam ml 10,000
24 ng nghim thy tinh ng 1,000
25 Bt vit knh Ci 0,020
26 Bt bi Ci 0,010
27 Bt la Ci 0,010
28 S lu kt qu xt nghim T 0,001
29 Cn st trng tay nhanh ml 1,000
30 Dung dch nc ra tay ml 8,000
31 Khn lau tay Ci 0,030
32 Giy tr kt qu xt nghim T 2,000
33 QC (nu thc hin) * 0,1
34 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
-
3
Cc loi bnh phm c ch nh xt nghim vi khun.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
- Nh dung dch tm gentian, ph kn ni dn phin, duy tr 1 - 2 pht
- dung dch tm gentian, ra tiu bn di vi nc chy nh
- Nh dung dch lugol, 30 giy
- dung dch lugol, ra nc
- Ty mu: nh vi git cn 95% ln tiu bn, nghing i nghing li cho
cn chy t cnh n sang cnh kia. Khi thy mu tm trn lam knh va phai
ht th ra nc ngay.
- Nh dung dch fuchsin, 1 - 2 pht
- Ra nc k, kh tiu bn, soi knh hin vi.
IV. NHN NH KT QU
Ch c kt qu khi QC t tiu chun.
1. nh gi hnh nh vi khun trn tiu bn nhum Gram
- Soi di vt knh du (x100)
- Din gii v c kt qu:
Vi khun Gram (+) bt mu tm sm ca gentian.
Vi khun Gram (-) bt mu ca fuchsin.
2. nh gi hnh nh t bo trn tiu bn nhum n (nu c).
V. NHNG SAI ST V X TR
Gram dng gi: Ty cn cha thi gian
Gram m gi:
+ Tui ca mu cy vi khun nh hng ln tnh cht nhum Gram cc mu cy vi khun thi gian qu lu.
+ Ty cn qu lu v trng khng k.
Nhum li tiu bn khi nghi ng kt qu khng chnh xc.
-
4
2. Vi khun test nhanh
I. MC CH V NGUYN L
1. Mc ch
Xc nh nhanh s c mt ca vi khun gy bnh trong bnh phm thng
qua pht hin s c mt ca khng nguyn vi khun.
2. Nguyn l
Pht hin khng nguyn vi khun da trn phn ng kt hp c hiu
gia khng nguyn vi khng huyt thanh mu.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1 . Trang thit b
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Bng Kg 0,001
2 Dy gar Ci 0,001
3 Cn ml 1,000
4 Bm kim tim Ci 1,000
5 Panh Ci 0,0001
6 Khay ng bnh phm Ci 0,0001
7 Hp vn chuyn bnh phm Ci 0,001
8 Tube ng bnh phm Ci 1,000
9 Sinh phm chn on Test 1,000
10 u cn 200 ul Ci 2,000
11 Giy thm Cun 0,100
12 Giy xt nghim T 2,000
13 S lu kt qu xt nghim T 0,001
-
5
14 Bt vit knh Ci 0,020
15 Bt bi Ci 0,010
16 M Ci 0,020
17 Khu trang Ci 0,020
18 Gng tay i 0,100
19 Gng tay x l dng c i 0,020
20 Qun o bo h B 0,005
21 Dung dch nc ra tay ml 8,000
22 Cn st trng tay nhanh ml 1,000
23 Dung dch kh trng ml 10,000
24 Khn lau tay Ci 0,010
25 QC (nu thc hin) * 0,1
26 EQAS (nu thc hin) * 0,005
Thc hin xt nghim 05 mu bnh phm/ln.
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Cc loi bnh phm ch nh lm test nhanh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu
III. CC BC THC HIN
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1).
2. Tin hnh k thut
B sinh phm Slidex Meningite Kit 5 (VD) pht hin 5 cn nguyn vi
khun gy vim mng no.
-
6
Cc bc
thc hin
K thut
1. a sinh phm ra ngoi nhit phng
nh s th t cc bnh phm v s th t trn tm knh
2. Lc nh nhng l cha ht latex, khng c lc qu mnh
3. Dng pipet nh 1 git dch no ty ca ngi bnh vo tng ng nh s trn phin knh. Nh chng dng v
chng m
4. Lc nh nhng l c cha ht latex v nh mt git vo cnh git dch no ty ca ngi bnh
5. Trn u 2 loi vi nhau bng que trn ph u b mt ca mi .
6. Lc u c phin knh bng tay hoc dng my lc 80-100 vng trong 10 pht
7. c kt qu
IV. NHN NH KT QU
- chng dng: c nhin tng ngng kt ht latex
- chng m: khng c nhin tng ngng kt, hn dch nhn thy mn, ng nht
- khng c hin tng ngng kt, hn dch nhn thy mn, ng nht: m tnh
- c hin tng ngng kt, hn dch nhn thy th, c ht ngng kt r trn nn en.
V. NHNG SAI ST V X TR
Lng bnh phm a vo qu nhiu hoc qu t u c th nh hng
n kt qu.
Do vy, phi tin hnh ly th tch bnh phm ng theo yu cu (Xem
Ph lc 6).
-
7
3. Vi khun nui cy v nh danh
phng php thng thng
I. MC CH V NGUYN L
1. Mc ch
Pht hin v nh danh vi khun gy bnh bng phng php nui cy
kinh in.
2. Nguyn l
Vi khun c nh danh da vo c im nui cy, mt s tnh cht
chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng
nguyn.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
3. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1 .Trang thit b
- T an ton sinh hc cp 2
- Knh hin vi quang hc
- T m thng
- T m CO2
- My tnh ci phn mm c API (nu c)
2.2 .Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Que cy Ci 1,000
3 Lam knh Ci 2,000
4 Du soi knh ml 1,000
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
-
8
7 Thuc nhum fuchsin ml 5,000
8 Thuc nhum tm gentian ml 5,000
9 Cn ty 95 ml 10,000
10 Lugol ml 5,000
11 Thuc nhum xanh methylen ml 5,000
12 Mi trng nui cy a 1,500
13 B sinh vt ha hc (b API ) Test 0,500
14 Bng Kg 0,001
15 Cn 90 (v sinh dng c) ml 10,000
16 n cn Ci 0,0001
17 Panh Ci 0,0001
18 Khay ng bnh phm Ci 0,0001
19 Hp vn chuyn bnh phm Ci 0,0001
20 M Ci 0,020
21 Khu trang Ci 0,020
22 Gng tay i 3,000
23 Gng tay x l dng c i 0,020
24 Qun o bo h B 0,001
25 Acid ngm lam ml 10,000
26 ng nghim thy tinh ng 1,000
27 Bt vit knh Ci 0,020
28 Bt bi Ci 0,010
29 Bt la Ci 0,010
30 S lu kt qu xt nghim T 0,001
31 Cn st trng tay nhanh ml 1,000
32 Dung dch nc ra tay ml 8,000
33 Khn lau tay Ci 0,030
34 Giy tr kt qu xt nghim T 2,000
35 QC (nu thc hin) * 0,1
36 EQAS (nu thc hin) * 0,005
Mi trng nui cy v ha cht nh danh vi khun c tnh trn t l
dng tnh trung bnh l 50 % cho cc loi bnh phm.
-
9
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1).
2. Tin hnh k thut
- Nhum soi bnh phm, nh gi s b
- Nui cy bnh phm vo mi trng phn lp
- m qua m
- Bt khun lc nghi ng
- Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc (b API )
IV. NHN NH KT QU
- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.
- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy,
khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c
ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn
nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny.
Nu c yu cu tm cn nguyn vi khun him gp, phi ghi yu cu c
th.
Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a
n kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).
-
10
4. Vi khun nui cy, nh danh h thng t ng
I. MC CH V NGUYN L
1. Mc ch
Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn
h thng t ng.
2. Nguyn l
Vi khun c nh danh bng my t ng da trn mt s tnh cht
chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- Knh hin vi quang hc
- T m thng
- T m CO2
- H thng nh danh t ng
- My o c
- L hp t
- T an ton sinh hc cp 2
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Que cy Ci 2,000
3 Lam knh Ci 2,000
4 Du soi knh ml 1,000
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
-
11
7 Thuc nhum fuchsin ml 5,000
8 Thuc nhum tm gentian ml 5,000
9 Cn ty 95 ml 10,000
10 Lugol ml 5,000
11 Thuc nhum xanh methylen ml 5,000
12 Mi trng nui cy a 1,500
13 Panel nh danh theo h thng my t ng Test 0,500
14 B Panel chng B 0,0002
15 Bm kim tim Ci 1,000
16 Bng Kg 0,001
17 Cn 90 (v sinh dng c) ml 10,000
18 n cn Ci 0,0001
19 Panh Ci 0,0001
20 Khay ng bnh phm Ci 0,0001
21 Hp vn chuyn bnh phm Ci 0,0001
22 M Ci 0,020
23 Khu trang Ci 0,020
24 Gng tay i 3,000
25 Gng tay x l dng c i 0,020
26 Qun o bo h B 0,001
27 Acid ngm lam ml 10,000
28 ng nghim thy tinh ng 1,000
29 Bt vit knh Ci 0,020
30 Bt bi Ci 0,010
31 Bt la Ci 0,010
32 S lu kt qu xt nghim T 0,001
33 Cn st trng tay nhanh ml 1,000
34 Dung dch nc ra tay ml 8,000
35 Khn lau tay Ci 0,030
36 Giy tr kt qu xt nghim T 2,000
37 QC (nu thc hin) * 0,1
38 EQAS (nu thc hin) * 0,005
-
12
Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s
bnh phm gi xt nghim.
B Panel s dng kim chun c tnh trn 5000 Test/1 nm.
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
II. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1).
2. Tin hnh k thut
- Nhum soi bnh phm, nh gi s b
- Nui cy bnh phm vo mi trng phn lp
- m qua m
- Bt khun lc nghi ng
- Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng.
IV. NHN NH KT QU
- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.
- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy,
khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c
ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn
nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny.
Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th.
-
13
Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n
kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).
-
14
5. Vi khun nui cy, nh danh v khng thuc h thng
t ng
I. MC CH V NGUYN L
1. Mc ch
Pht hin vi khun gy bnh bng phng php nui cy, nh danh v
khng thuc trn h thng t ng.
2. Nguyn l
Vi khun c nh danh bng my t ng da trn mt s tnh cht
chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu; v
xc nh mc nhy cm vi khng sinh bng phng php khng sinh pha
long.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- Knh hin vi quang hc
- T m thng
- T m CO2
- H thng nh danh t ng
- My o c
- L hp t
- T an ton sinh hc cp 2
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Lam knh Ci 2,000
3 Que cy Ci 2,000
4 Du soi knh ml 1,000
-
15
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
7 Thuc nhum Fuchsin ml 5,000
8 Thuc nhum tm Gentian ml 5,000
9 Cn ty 95 ml 10,000
10 Lugol ml 5,000
11 Thuc nhum xanh methylen ml 5,000
12 Mi trng nui cy a 1,500
13 Panel nh danh theo h thng my t ng Test 1,000
14 B Panel chng B 0,0002
15 Bm kim tim Ci 1,000
16 Bng Kg 0,001
17 Cn 90 (v sinh dng c) ml 10,000
18 n cn Ci 0,0001
19 Panh Ci 0,0001
20 Khay ng bnh phm Ci 0,0001
21 Hp vn chuyn bnh phm Ci 0,0001
22 M Ci 0,020
23 Khu trang Ci 0,020
24 Gng tay i 3,000
25 Gng tay x l dng c i 0,020
26 Qun o bo h B 0,001
27 Acid ngm lam ml 10,000
28 ng nghim thy tinh ng 1,000
29 Bt vit knh Ci 0,020
30 Bt bi Ci 0,010
31 Bt la Ci 0,010
32 S lu kt qu xt nghim T 0,001
33 Cn st trng tay nhanh ml 1,000
34 Dung dch nc ra tay ml 8,000
35 Khn lau tay Ci 0,030
36 Giy tr kt qu xt nghim T 2,000
-
16
37 QC (nu thc hin) * 0,1
38 EQAS (nu thc hin) * 0,005
Mi trng nui cy c tnh trn t l dng l 50 % so vi tng s
bnh phm gi xt nghim.
B Panel s dng kim chun c tnh trn 5000 Test/1 nm.
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1).
2. Tin hnh k thut
- Nhum soi bnh phm, nh gi s b
- Nui cy bnh phm vo mi trng phn lp
- m qua m
- Bt khun lc nghi ng
- Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng.
IV. NHN NH KT QU
- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.
- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy,
khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c
-
17
ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn
nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny.
S dng sai panel nh danh cho loi vi khun cn xc nh. Cn chn la
chnh xc panel nh danh cho tng loi vi khun.
Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th.
Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n
kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).
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18
6. Vi khun khng thuc nh tnh
I. MC CH V NGUYN L
1. Mc ch
Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy
bnh.
2. Nguyn l
Mc nhy cm vi khng sinh ca cc chng vi khun th nghim
c nh gi bng phng php khng sinh khuch tn trong thch. S pht
trin ca vi khun s b c ch khi khng sinh t n mt nng nht nh.
ng knh vng c ch t l thun vi mc nhy cm c phin gii ra
cc phn loi S (sensitive nhy cm), I (intermediate trung gian), hoc R
(resistant khng) khi so snh vi bng chun CSLI cp nht hng nm.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1 . Trang thit b
- T m
- T an ton sinh hc cp 2
- My lc
- ng c chun McFarland 0.5
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Khoanh giy khng sinh Khoanh 22,000
2 Que cy Ci 1,000
3 Thch Muller Hinton a 3,000
4 Thch mu a 1,000
5 Giy khng sinh T 2,000
6 Bng Kg 0,001
7 Cn 90 (v sinh dng c) ml 10,000
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19
8 Panh Ci 0,0001
9 Khay ng bnh phm Ci 0,0001
10 Hp vn chuyn bnh phm Ci 0,0001
11 M Ci 0,050
12 Khu trang Ci 0,050
13 Gng tay i 3,000
14 Gng tay x l dng c i 0,030
15 Qun o bo h B 0,001
16 Nc mui sinh l ml 5,000
17 ng nghim thy tinh ng 1,000
18 Tm bng v trng Ci 1,000
19 Bt vit knh Ci 0,020
20 Bt bi Ci 0,010
21 Bt la Ci 0,010
22 S lu kt qu xt nghim Quyn 0,001
23 Cn st trng tay nhanh ml 1,000
24 Dung dch nc ra tay ml 8,000
25 Khn lau tay Ci 0,050
26 QC (nu thc hin) * 0,1
27 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
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20
1. Ly bnh phm
Chng vi khun cn th nghim c nui cy thun nht trong iu
kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi).
2. Tin hnh k thut
2.1. Pha huyn dch vi khun
Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin
u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch
ng nht. So snh c ca huyn dch vi khun vi c ca ng
McFarland 0,5.
2.2 . Dn u canh khun ln mt a thch
Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn,
p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun
loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u
que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c
dn u ln trn ton b b mt a thch
2.3. t cc khoanh giy khng sinh ln mt thch
Cc khoanh giy sau khi t cn c n xung va phi m bo chng
c tip xc hon ton vi mt thch. Trong vng 15 pht sau khi t khoanh
giy khng sinh, cc a thch phi c lt p trong t m 35C.
2.4. a thch trong t m 16 - 24 gi
IV. NHN NH KT QU
Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.
o ng knh vng c ch (bao gm c ng knh ca khoanh giy
khng sinh) tnh theo mm.
Phin gii ng knh vng c ch ra kt qu S, I, R theo hng dn ca
CLSI cp nht hng nm.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d
nui cy, khng p dng cho cc vi khun k kh bt buc.
Sai st c th gp khi:
- Ln hai hay nhiu chng vi khun
- Vi khun mc qu dy hoc qu tha.
Phi tin hnh lm li khi thy cc hin tng nh trn.
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21
7. Vi khun khng thuc h thng t ng
I. MC CH V NGUYN L
1. Mc ch
Xc nh mc nhy cm vi khng sinh ca cc chng vi khun gy
bnh.
2. Nguyn l
Mc nhy cm vi khng sinh ca cc chng vi khun th nghim
c nh gi da trn gi tr ca nng c ch ti thiu - MIC. Gi tr MIC
s c phin gii ra phn loi S (sensitive nhy cm), I (intermediate trung
gian), hoc R (resistant khng) nh h thng t ng.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1 . Trang thit b
- My nh danh, khng sinh t ng (v d H thng Vitek 2 Compact)
- T an ton sinh hc cp 2
- My vi tnh
- My in
2.2 . Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Th khng sinh Th 1,000
2 Que cy Ci 1,000
3 Blood agar base 500G ml 75,000
4 Dng c ly bnh phm L 1,000
5 Mac Conkey agar ml 75,000
6 Khay ng bnh phm Ci 0,0001
7 M Ci 0,030
8 Khu trang Ci 0,030
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22
9 Gng tay i 3,000
10 Gng tay x l dng c i 0,030
11 Qun o bo h Ci 0.0005
12 ng nghim thy tinh ng 1,000
13 Dung dch nc ra tay ml 8,000
14 Khn lau tay Ci 0,030
15 Giy tr kt qu xt nghim T 4,000
16 Presept Vin 1,000
17 QC (nu thc hin) * 0,1
18 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Chng vi khun cn th nghim c nui cy thun nht trong iu
kin ti u v ang giai on pht trin mnh (nui cy sau 16-24 gi).
2. Tin hnh k thut
- Chun b mu
- Np ng nghim v card test ng loi vo cassette
- Nhp mu vo my
- Khai bo thng tin
- To thng tin bnh nhn
- Xem kt qu ca mu
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23
IV. NHN NH KT QU
Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.
Ghi nhn cc kt qu S, I hay R cng vi gi tr MIC.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d
nui cy, khng p dng cho cc vi khun k kh bt buc.
Sai st c th gp khi:
- Ln hai hay nhiu chng vi khun
- Chn th lm khng sinh khng ph hp vi chng vi khun cn th
nghim
- Chng vi khun sau khi pha khng c a vo my trong vng 30 pht
Phi tin hnh lm li khi thy cc hin tng nh trn.
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24
8. Vi khun khng thuc nh lng MIC
(cho 1 loi khng sinh)
I. MC CH V NGUYN L
1. Mc ch
Xc nh mc nhy cm v nng c ch ti thiu ca khng sinh
vi chng vi khun gy bnh.
2. Nguyn l
Etest l thanh plastic kch thc 5 x 57 mm c phn b di MIC theo
g/mL v c m code tn khng sinh. C 15 nng khng sinh c pha
long bc 2 gn c nh mt mt ca bng plastic. Khi bng Etest c t ln
mt thch dn vi khun, khng sinh c bc nng nhanh chng khuch
tn trong thch. Sau khi nui cy qua m, s xut hin vng c ch hnh elip
i xng qua thanh plastic. Gi tr MIC c xc nh trc tip ti im ct ca
hnh elip vi thanh Etest. Gi tr MIC s c phin gii ra phn loi S
(sensitive nhy cm), I (intermediate trung gian), hoc R (resistant
khng) khi so snh vi bng chun CLSI cp nht hng nm.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1 . Trang thit b
- T m
- T an ton sinh hc cp 2
- My lc
- ng c chun McFarland 0.5
2.2 . Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Thanh giy khng sinh Ci 1,000
2 Que cy Ci 1,000
3 Thch Muller Hinton a 1,000
4 Thch mu a 1,000
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25
5 Giy khng sinh T 2,000
6 Bng Kg 0,001
7 Cn 90 (v sinh dng c) ml 10,000
8 Panh Ci 0,0001
9 Khay ng bnh phm Ci 0,0001
10 Hp vn chuyn bnh phm Ci 0,0001
11 M Ci 0,050
12 Khu trang Ci 0,050
13 Gng tay i 3,000
14 Gng tay x l dng c i 0,030
15 Qun o bo h B 0,001
16 Nc mui sinh l ml 5,000
17 ng nghim thy tinh ng 1,000
18 Tm bng v trng Ci 1,000
19 Bt vit knh Ci 0,020
20 Bt bi Ci 0,010
21 Bt la Ci 0,010
22 S lu kt qu xt nghim Quyn 0,001
23 Cn st trng tay nhanh ml 1,000
24 Dung dch nc ra tay ml 8,000
25 Khn lau tay Ci 0,050
26 QC (nu thc hin) * 0,1
27 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3 Bnh phm
Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi.
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26
4 Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Chng vi khun cn th nghim c nui cy thun nht trong iu kin ti
u v ang giai on pht trin mnh (nui cy sau 16-24 gi).
2. Tin hnh k thut
2.1. Pha huyn dch vi khun
Dng que cy ly vi khun t 3-5 khun lc c hnh thi ging nhau nghin
u vo ng nc mui sinh l 5 ml, lc u trn my lc c huyn dch
ng nht. So snh c ca huyn dch vi khun vi c ca ng
McFarland 0,5.
2.2. Dn u canh khun ln mt a thch
Dng tm bng v trng nhng vo ng huyn dch vi khun pha trn,
p nh v xoay trn tm bng trn thnh bn ca ng huyn dch vi khun
loi bt phn huyn dch vi khun thm vo u tm bng. Sau , ria u
que tm bng trn ton b mt a thch Mueller-Hinton sao cho vi khun c
dn u ln trn ton b b mt a thch
2.3. t cc khoanh giy khng sinh ln mt thch
t thanh Etest ln mt thch sao cho mt c ghi di nng hng ln trn
v phi m bo ton b b mt ca thanh Etest c tip xc hon ton vi
mt thch. Khi t xong thanh Etest khng c dch chuyn thanh Etest
khi v tr.
2.4. a thch trong t m 16 - 24 gi
IV. NHN NH KT QU
1. Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.
2. c kt qu
Sau m 16 24 h v khi thy r vi khun mc, c gi tr MIC im ct
ca hnh elip vi thanh E-test. Lm trn gi tr MIC im gia hai bc pha
long ln gi tr cao hn mt bc trc khi phin gii kt qu.
Phin gii kt qu MIC ra gi tr S, I hoc R theo ti liu ca CLSI cp
nht hng nm.
Vi cc khng sinh dit khun, phi c gi tr MIC ti im vi khun b
c ch hon ton. Vi cc khng sinh km khun, phi c gi tr MIC ti im
vi khun b c ch 80%.
V. NHNG SAI ST V X TR
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27
Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d
nui cy, khng p dng cho cc vi khun k kh bt buc.
Sai st c th gp khi:
- Ln hai hay nhiu chng vi khun
- Vi khun mc qu dy hoc qu tha.
Phi tin hnh lm li khi thy cc hin tng nh trn.
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28
9. Vi khun khng sinh phi hp
I. MC CH V NGUYN L
1. Mc ch
Tm cp khng sinh phi hp c tc dng hip ng trn chng vi khun
gy bnh.
2. Nguyn l
Xc nh tc dng hip ng, i khng hoc khng khc bit (gi tr
MIC) khi th nghim phi hp 2 khng sinh khc nhau vi vi khun gy bnh
bng thanh E-test da trn ch s FIC (Fractional Inhibitory Concentration).
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
2.1. Trang thit b
- T m
- T an ton sinh hc cp 2
- My lc
- ng c chun McFarland 0.5
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Thanh giy khng sinh Ci 4,000
2 Thch Muller Hinton a 3,000
3 Que cy Ci 1,000
4 Thch mu a 2,000
5 Giy khng sinh T 2,000
6 Bng Kg 0,001
7 Cn 90 (v sinh dng c) ml 10,000
8 Panh Ci 0,0001
9 Khay ng bnh phm Ci 0,0001
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29
10 Hp vn chuyn bnh phm Ci 0,0001
11 M Ci 0,050
12 Khu trang Ci 0,050
13 Gng tay i 3,000
14 Gng tay x l dng c i 0,030
15 Qun o bo h bo h B 0,001
16 Nc mui sinh l ml 5,000
17 ng nghim thy tinh ng 1,000
18 Tm bng v trng Ci 1,000
19 Bt vit knh Ci 0,020
20 Bt bi Ci 0,010
21 Bt la Ci 0,010
22 S lu kt qu xt nghim Quyn 0,001
23 Cn st trng tay nhanh ml 1,000
24 Dung dch nc ra tay ml 8,000
25 Khn lau tay Ci 0,050
26 QC (nu thc hin) * 0,1
27 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Chng vi khun c xc nh l cn nguyn gy bnh, thun v mi.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
- Pha huyn dch vi khun c c tng ng vi c McFarland 0.5
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30
- Dn u canh khun ln mt 2 a thch Mueller-Hinton v cy thun trn 1 a thch mu
- t 2 thanh E-test ln 2 a thch dn u vi khun
- cc a thch trong t m 16 - 18 gi
- c kt qu gi tr MIC ca c hai khng sinh
- Pha huyn dch vi khun c c tng ng vi c McFarland 0.5 ri dn u canh khun ln mt 1 a thch Mueller-Hinton v cy thun trn 1
a thch mu
- t 2 thanh E-test vung gc vi nhau, giao nhau ti gi tr MIC ca mi thanh c bc 5 ln a thch dn u vi khun
- cc a thch trong t m 16 - 18 gi
- c kt qu gi tr MIC ca c hai khng sinh
IV. NHN NH KT QU
Ch c kt qu khng sinh chng ngi bnh khi kt qu QC t.
c kt qu:
- Tnh ch s FIC (Fractional Inhibitory Concentration)
FIC ca khng sinh A = MIC ca khng sinh A trong th nghim phi
hp/MIC ca khng sinh A th nghim ring
FIC ca khng sinh B = MIC ca khng sinh B trong th nghim phi
hp/MIC ca khng sinh B th nghim ring
Ch s FIC = FIC ca khng sinh A/ FIC ca khng sinh B
- Phin gii kt qu ch s FIC
o Hai khng sinh c tc dng hip ng nu ch s FIC 0.5
o Hai khng sinh c tc dng i khng nu ch s FIC > 4
o Hai khng sinh phi hp khng c tc dng khc bit nu ch s FIC 4 v > 0.5
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng cho cc chng vi khun hiu k kh ty tin d
nui cy, khng p dng cho cc vi khun k kh bt buc.
Nhng trng hp tc dng i khng yu c th khng pht hin c v
vng c ch nm di vng bt cho ca hai thanh E-test.
Sai st c th gp khi:
- Ln hai hay nhiu chng vi khun
- Vi khun mc qu dy hoc qu tha.
- Phi tin hnh lm li khi thy cc hin tng nh trn.
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31
10. Vi khun k kh nui cy v nh danh
I. MC CH V NGUYN L
1. Mc ch
Pht hin v nh danh vi khun k kh gy bnh bng phng php nui
cy kinh in.
2. Nguyn l
Vi khun c nh danh da vo c im nui cy, mt s tnh cht
chuyn ha, cc c im v hnh thi hc v c th kt hp vi tnh cht khng
nguyn.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- Knh hin vi quang hc
- T an ton sinh hc cp 2
- H thng my nui cy vi khun k kh
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Lam knh Ci 2,000
3 Que cy Ci 2,000
4 Du soi knh ml 1,000
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
7 Thuc nhum fuchsin ml 5,000
8 Thuc nhum tm gentian ml 5,000
9 Cn ty 95 ml 10,000
10 Lugol ml 5,000
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32
11 Thuc nhum xanh methylen ml 5,000
12 Mi trng nui cy k kh chai 1,000
13 Mi trng thch mu thng a 2,000
14 Mi trng thch mu k kh a 2,000
15 B gi ng API Test 1,000
16 Bnh kh trn (tnh theo gi) Gi 168,000
17 Bm kim tim Ci 1,000
18 Bng Kg 0,001
19 Cn 90 (v sinh dng c) ml 10,000
20 n cn Ci 0,0001
21 Panh Ci 0,0001
22 Khay ng bnh phm Ci 0,0001
23 Hp vn chuyn bnh phm Ci 0,0001
24 M Ci 0,020
25 Khu trang Ci 0,020
26 Gng tay i 3,000
27 Gng tay x l dng c i 0,020
28 Qun o bo h bo h B 0,001
29 Acid ngm lam ml 10,000
30 ng nghim thy tinh ng 1,000
31 Bt vit knh Ci 0,020
32 Bt bi Ci 0,010
33 Bt la Ci 0,010
34 S lu kt qu xt nghim T 0,001
35 Cn st trng tay nhanh ml 1,000
36 Dung dch nc ra tay ml 8,000
37 Khn lau tay Ci 0,030
38 Giy tr kt qu xt nghim T 2,000
39 QC (nu thc hin) * 0,1
39 EQAS (nu thc hin) * 0,005
* Ghi ch:
-
33
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Tt c cc loi bnh phm c ch nh nui cy tm vi khun k kh gy bnh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem Ph lc 1).
2. Tin hnh k thut
- Nhum soi bnh phm, nh gi s b
- Nui cy bnh phm vo mi trng phn lp
- m qua m
- Bt khun lc nghi ng
- Nhum soi, th nghim cc th nghim sinh vt ha hc n gin v nh danh bng cc b sinh vt ha hc chuyn dng.
- Tt c cc bc tin hnh v m u c thc hin bn trong h thng my nui cy vi khun k kh nhm m bo iu kin mi trng k kh.
IV. NHN NH KT QU
- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.
- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng tm vi khun k kh bt buc v k kh ty tin.
Kt qu m tnh khng c ngha l khng c vi khun gy bnh trong bnh
phm m l khng tm thy cn nguyn vi khun gy bnh c th phn lp c
bng quy trnh nui cy ny.
Tt c cc thao tc k thut phi din ra bn trong h thng my to mi
trng k kh m bo iu kin nui cy thch hp cho vi khun k kh mc.
Kh trng s dng nui cy vi khun phi m bo ng nng .
Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th.
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34
11. Vi khun khng nh
I. MC CH V NGUYN L
1. Mc ch
Pht hin vi khun gy bnh bng phng php nui cy, nh danh trn
h thng t ng.
2. Nguyn l
Vi khun c nh danh bng my t ng da trn mt s tnh cht
chuyn ha kt hp vi cc c im v hnh thi hc v tnh cht bt mu.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- Knh hin vi quang hc
- T m thng
- T m CO2
- H thng nh danh t ng
- My o c
- L hp t
- T an ton sinh hc cp 2
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 L ly bnh phm L 1,000
2 Lam knh Ci 2,000
3 Que cy Ci 1,000
4 Du soi knh ml 1,000
5 Cn 96 lau knh ml 1,000
6 Nc mui sinh l ml 5,000
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35
7 Thuc nhum Fuchsin ml 5,000
8 Thuc nhum tm Gentian ml 5,000
9 Cn ty 95 ml 10,000
10 Lugol ml 5,000
11 Thuc nhum xanh methylen ml 5,000
12 Mi trng nui cy a 3,000
13 Panel nh danh theo h thng my t ng Test 1,000
14 B Panel chng B 0,0002
15 Bm kim tim Ci 1,000
16 Bng Kg 0,001
17 Cn 90 (v sinh dng c) ml 10,000
18 n cn Ci 0,0001
19 Panh Ci 0,0001
20 Khay ng bnh phm Ci 0,0001
21 Hp vn chuyn bnh phm Ci 0,0001
22 M Ci 0,020
23 Khu trang Ci 0,020
24 Gng tay i 3,000
25 Gng tay x l dng c i 0,020
26 Qun o bo h B 0,001
27 Acid ngm lam ml 10,000
28 ng nghim thy tinh ng 1,000
29 Bt vit knh Ci 0,020
30 Bt bi Ci 0,010
31 Bt la Ci 0,010
32 S lu kt qu xt nghim T 0,001
33 Cn st trng tay nhanh ml 1,000
34 Dung dch nc ra tay ml 8,000
35 Khn lau tay Ci 0,030
36 Giy tr kt qu xt nghim T 2,000
37 QC (nu thc hin) * 0,1
37 EQAS (nu thc hin) * 0,005
-
36
B Panel s dng kim chun c tnh trn 5000 Test/1 nm.
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Tt c cc loi bnh phm c ch nh nui cy tm vi khun gy bnh.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 1).
2. Tin hnh k thut
- Nhum soi bnh phm, nh gi s b
- Nui cy bnh phm vo mi trng phn lp
- m qua m
- Bt khun lc nghi ng
- Nhum soi, th nghim cc tnh cht sinh vt ha hc n gin v nh danh bng h thng t ng.
IV. NHN NH KT QU
- Dng tnh: Phn lp v nh danh c vi khun gy bnh. Tr kt qu tn vi khun n mc chi v/hoc loi.
- m tnh: Khng tm thy hoc khng phn lp c vi khun gy bnh.
V. NHNG SAI ST V X TR
Quy trnh ny ch p dng tm vi khun hiu k kh ty tin d nui cy,
khng p dng cho cc vi khun k kh bt buc. Kt qu m tnh khng c
ngha l khng c vi khun gy bnh trong bnh phm m l khng tm thy cn
nguyn vi khun gy bnh c th phn lp c bng quy trnh nui cy ny.
Nu c yu cu tm cn nguyn vi khun no , phi ghi yu cu c th.
Bnh phm ly, vn chuyn v bo qun khng ng yu cu c th a n
kt qu m tnh hoc dng tnh gi (Xem Ph lc 6).
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37
12. Vi khun nh danh PCR
I. MC CH V NGUYN L
1. Mc ch
Chn on nhanh vi khun trc tip t bnh phm.
2. Nguyn l
Xc nh s c mt ca gene c trng cho vi khun bng k thut PCR.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- T an ton Sinh hc cp 2
- My PCR
- My in di
- My c in di
- My ly tm 25000 x g
- My nhit
- My vortex
- My ly tm dng cho tube 0,2 ml
- Micropipette
- T m su (-200C)
- B lu in.
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
nh mc sinh phm v vt t tiu hao cho 2 mu/ln thc hin (VD)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Bng kg 0,001
2 Cn ml 1,000
3 Bm kim tim Ci 1,000
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38
4 Panh Ci 0,0001
5 Khay ng bnh phm Ci 0,0001
6 Hp vn chuyn bnh phm Test 0,001
7 Tube ng bnh phm Ci 2,000
8 Gng khng c bt (DNase-RNase free) Ci 0,500
9 Sinh phm chn on Test 1,000
10 Khu hao sinh phm cho kim tra li Test 1,350
11 Kit tch chit DNA Test 2,350
12 DNA marker B 1,000
13 Primer 1 ml 0,0001
14 Primer 2 ml 0,0001
15 Ependoff 1,7ml Tube 3,000
16 Ependoff 0,2ml Tube 1,000
17 u cn 10 l c lc Ci 1,000
18 u cn 30 l Ci 1,200
19 u cn 200 l c lc Ci 2,200
20 u cn 1 ml c lc Ci 3,200
21 Ethanol BDH ml 0,500
22 Water-DEPC Treated ml 2,000
23 Giy thm Cun 0,100
24 Giy xt nghim T 2,000
25 S lu kt qu xt nghim T 0,001
26 Bt vit knh Ci 0,020
27 Bt bi Ci 0,010
28 M Ci 0,020
29 Khu trang Ci 0,020
30 Gng tay i 0,100
31 Gng tay x l dng c i 0,020
32 Qun o bo h B 0,005
33 Dung dch nc ra tay ml 8,000
34 Cn st trng tay nhanh ml 1,000
35 Dung dch kh trng ml 10,000
-
39
36 Khn lau tay ci 0,010
37 QC (nu thc hin) * 0,100
38 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Vi khun thun v bnh phm phn lp
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu.
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 3).
2. Tin hnh k thut
- Tch chit DNA tng s
- Thc hin PCR
- in di kim tra sn phm
- nh gi v kt lun
IV. NHN NH KT QU
Sn phm PCR phi c mt bng c hiu duy nht, r nt, khng b t gy
v c kch thc tng ng vi on gene ch cn khuch i.
V. NHNG SAI ST V X TR
Khi kt qu khi chng m v/hoc chng dng khng hp l, cn phi
kim tra li qu trnh tch chit DNA tng s, cht lng primers v master
mix, sau thc hin li xt nghim.
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40
13. Vi khun nh danh gii trnh t gene
I. NGUYN L
1. Mc ch
nh danh mi cn nguyn vi khun gy bnh n mc chi v hoc
loi.
2. Nguyn l
nh danh chnh xc cc loi vi khun da trn trnh t nucleotide c
trng ca gene m ha cho 16S rRNA ca vi khun bng k thut gii trnh t
gene.
II. CHUN B
1.Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- T an ton Sinh hc cp 2
- My PCR
- My c in di
- My gii trnh t gen
- My ly tm 25000 x g
- My nhit
- My vortex
- My ly tm dng cho tube 0,2 ml
- Micropipette
- B lu in
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
nh mc sinh phm v vt t tiu hao cho 3 mu/ln thc hin (VD)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Bng kg 0,001
2 Cn ml 1,000
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41
3 Bm kim tim Ci 1,000
4 Panh Ci 0,0001
5 Khay ng bnh phm Ci 0,0001
6 Hp vn chuyn bnh phm Test 0,001
7 Tube ng bnh phm Ci 2,000
8 Gng khng c bt (DNase-RNase free) Ci 0,500
9 Sinh phm chn on Test 1,000
10 Khu hao sinh phm cho kim tra li Test 1,350
11 Kit tch chit DNA Test 2,350
12 DNA marker B 1,000
13 Primer 1 (b primer c hiu) ml 0,0001
14 Primer 2 (b primer c hiu) ml 0,0001
15 Primer 3 (b primer c hiu) ml 0,0001
16 Primer 4 (b primer c hiu) ml 0,0001
17 ng Eppendorf 1,5 ml Tube 3,000
18 ng Eppendorf 0,2 ml Tube 1,000
19 u cn 10 l c lc Ci 1,000
20 u cn 30 l Ci 1,200
21 u cn 200 l c lc Ci 2,200
22 u cn 1 ml c lc Ci 3,200
23 Ethanol BDH ml 0,500
24 Water-DEPC Treated ml 2,000
25 Giy thm Cun 0,100
26 Giy xt nghim T 2,000
27 S lu kt qu xt nghim T 0,001
28 Bt vit knh Ci 0,020
29 Bt bi Ci 0,010
30 M Ci 0,020
31 Khu trang Ci 0,020
32 Gng tay i 0,100
33 Gng tay x l dng c i 0,020
34 Qun o bo h B 0,005
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42
35 Dung dch nc ra tay ml 8,000
36 Cn st trng tay nhanh ml 1,000
37 Dung dch kh trng ml 10,000
38 Khn lau tay ci 0,010
39 QC (nu thc hin) * 0,100
40 EQAS (nu thc hin) * 0,005
* Ghi ch:
- Chi ph ni kim cho quy trnh k thut c tnh c th theo Chng trnh ni
kim (QC) l 1/10 tng chi ph dng c, ha cht, vt t tiu hao (vi s lng
10 mu cho 1 ln tin hnh k thut).
- Chi ph ngoi kim cho quy trnh k thut c tnh c th theo Chng trnh
ngoi kim (EQAS) l 1/200 tng chi ph dng c, ha cht, vt t tiu hao (vi
s ln ngoi kim trung bnh 2 ln/1 nm).
3. Bnh phm
Chng vi khun nui cy thun.
4. Phiu xt nghim
in y thng tin theo mu phiu yu cu
III. CC BC TIN HNH
Cc bc tin hnh thc hin theo phng tin, ha cht c v d trn.
1. Ly bnh phm
Theo ng quy nh ca chuyn ngnh Vi sinh (Xem ph lc 3).
2. Tin hnh k thut
- Tch chit DNA tng s
- Thc hin PCR gene 16S ARN ribosome
- in di kim tra sn phm
- Gii trnh t gene
- Kim tra tng ng DNA
IV. NHN NH KT QU
Sn phm PCR phi c mt bng c hiu duy nht, r nt v khng b
t gy. Trnh t DNA ca gene ch c so snh vi cc trnh t ca gene m
ha cho 16S rRNA c trn Genebank nh danh cn nguyn vi khun cn
xc nh. Trnh t nucleotide phi c tng ng 97 % mi c th kt lun
c n mc chi v/hoc loi.
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43
V. NHNG SAI ST V X TR
- Trong trng hp khng c sn phm PCR, cn phi kim tra li qu
trnh tch chit DNA tng s, cht lng primers v master mix, v thc hin
li.
- Nu trnh t DNA b nhiu cn phi kim tra li c hiu ca sn
phm PCR hoc qu trnh chy PCR sequencing b nhim cho.
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44
14. Vi khun khng thuc PCR
I. MC CH V NGUYN L
1. Mc ch
Pht hin c tnh khng thuc ca vi khun thng qua xc nh s c
mt ca gene khng thuc vi khun.
2. Nguyn l
S dng cp mi c trnh t c hiu xc nh s c mt ca gene
khng thuc trong vi khun gy bnh bng k thut PCR.
II. CHUN B
1. Ngi thc hin
- Ngi thc hin: Cn b xt nghim c o to v c chng ch hoc chng nhn v chuyn ngnh Vi sinh.
- Ngi nhn nh v ph duyt kt qu: cn b xt nghim c trnh i hc hoc sau i hc v chuyn ngnh Vi sinh.
2. Phng tin, ha cht
Phng tin, ha cht nh v d di y hoc tng ng.
2.1. Trang thit b
- T an ton Sinh hc cp 2
- My PCR
- My c in di
- My ly tm 25000 x g
- My nhit
- My vortex
- My ly tm dng cho tube 0,2 ml
- Micropipette
- B lu in.
2.2. Dng c, ha cht v vt t tiu hao (bao gm ni kim, ngoi kim)
nh mc sinh phm v vt t tiu hao cho 2 mu/ln thc hin (VD)
STT Chi ph ha cht, vt t tiu hao n v S lng
1 Bng kg 0,001
2 Cn ml 1,000
3 Bm kim tim Ci 1,000
4 Panh Ci 0,0001
5 Khay ng bnh phm Ci 0,0001
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45
6 Hp vn chuyn bnh phm Test 0,001
7 Tube ng bnh phm Ci 2,000
8 Gng khng c bt (DNase-RNase free) Ci 0,500
9 Sinh phm chn on Test 1,000
10 Khu hao sinh phm cho kim tra li Test 1,350
11 Kit tch chit DNA Test 2,350
12 DNA marker B 1,000
13 Primer 1 ml 0,0001
14 Primer 2 ml 0,0001
15 ng Eppendorf 1,5 ml Tube 3,000
16 ng Eppendorf 0,2 ml Tube 1,000
17 u cn 10 l c lc Ci 1,000
18 u cn 30 l Ci 1,200
19 u cn 200 l c lc Ci 2,200
20 u cn 1 ml c lc Ci 3,200
21 Ethanol BDH ml 0,500
22 Water-DEPC Treated ml 2,000
23 Giy thm Cun 0,100
24 Giy xt nghim T 2,000
25 S lu kt qu xt nghim T 0,001
26 Bt vit knh Ci 0,020
27 Bt bi Ci 0,010
28 M Ci 0,020
29 Khu trang Ci 0,020
30 Gng tay i 0,100
31 Gng tay x l dng c i 0,020
32 Qun o bo h B 0,005
33 Dung dch nc ra tay ml 8,000