comparative molecular-genetic characterization of collections of tubercle bacilli from distant...
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Comparative molecular-genetic characterization Comparative molecular-genetic characterization of collections of tubercle bacilli from distant of collections of tubercle bacilli from distant geographical regions of prime Soviet Uniongeographical regions of prime Soviet Union
Filipenko M.L., Rot M.A., Voronina E.N., Vikhrova M.A., Khrapov E.A., Ljashenko A.A., Krut’ko V.S., Poluektov E.I., Gorbunova V.V.,
Shabaldin A.V., Kushugulova A.R., Rakhimova S.E., Bekbolatova J.T.
Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk, RussiaKharkiv Medical Academy of Postgraduate Education, Phtiziatry and Pulmonology Department, Kharkiv, UkraineMLPU “Tuberculosis clinic”, Central Bacteriological Lab, Nizhnyi Novgorod, RussiaKemerovo Tuberculosis Clinic, Kemerovo, RussiaCancer Immunology Department, Kemerovo Scientific Center SB RAS, Kemerovo, RussiaRGKP “Republic Collection of Microorganisms”, Astana, Kazakhstan
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What kind of TB strains is What kind of TB strains is endemic for territory studiedendemic for territory studied
What kind of TB strains is What kind of TB strains is epidemic for territory studiedepidemic for territory studied
Directions of TB strains evolution Directions of TB strains evolution in clinical practice viewin clinical practice view
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Genotypic fingerprinting utilizes Genotypic fingerprinting utilizes slight differences in the total slight differences in the total
chromosome that could be or not chromosome that could be or not be related to phenotypic differencesbe related to phenotypic differences
..AGCATTCAG.. ..AGAATTCAG..
..CCTCGGGAAA
....CCCCGGGAAA..
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..GCTGGCTT…..AATTCCGAT..
..GCTAGCTT…..AATTCCGAT..
..GCTGGCTT…..AATTCTGAT..
..GCTAGCCT…..AATTCCGAT..
TIME
EVOLUTIONARY
RATE
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MethodsMethods
VNTR typingVNTR typing RFLP IS6110RFLP IS6110 RFLP Ser315Thr KatG RFLP Ser315Thr KatG RFLP Ser531Leu rpoB RFLP Ser531Leu rpoB Direct sequencing rpoBDirect sequencing rpoB
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Collections testedCollections tested
Kharkiv (Ukraine)Kharkiv (Ukraine) Astana (Kazakhstan)Astana (Kazakhstan) Nizhny Novgorod (Russia, West)Nizhny Novgorod (Russia, West) Kemerovo (Russia, East)Kemerovo (Russia, East)
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TypeTypeA,C,EA,C,E KazakhstanKazakhstan NovgorodNovgorod KharkivKharkiv KemerovoKemerovo
222222 6%6% 1, 4%1, 4% 23%23% 15%15%
243243 1%1% -- 5, 6%5, 6%
333333 -- -- 9%9% 4%4%
343343 4, 8%4, 8% -- 7%7%
345345 3, 6%3, 6% -- --
442442 -- -- 3%3%
443443 1%1% -- 4%4%
445445 41%41% 75%75% 24%24% 37%37%
452452 1%1% 1, 4%1, 4% 4%4%
455455 1%1% 1, 4%1, 4% --
totaltotal 5151 2323 8989 5454
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Древо филогенетического родства штаммов M.tuberculosis,
полученных из г.Харькова
0,00
0,05
0,10
0,15
0,20
0,25
0,30
0,35
0,40
31 126
94 56 46 29 67 51 122
105
102
98 78 76 75 74 61 44 41 38 37 36 35 27 24 23 11 7 14 32 84 15 12 125
42 40 43 79 69 64 52 13 123
115
110
109
103
100
92 86 85 83 71 65 62 57 53 34 17 10 58 128
113 54 127
63 45 101
47 26 18 9 811
2 49 87 22 48 20 124
90 19 5 4 73 21 6 3 2
222 452 445 333 343443 243
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38 41 44 73 74 75 76 85 86
222 222 445 243 222 222 222 445 445 А B A2
RFLP IS6110 analysis of TB strains from Kharkov
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mutmut wildwild
INHINHRR 4141 44 91%91%INHINHSS 44 3434
RIFRIFRR 3131 1919 62%62%RIFRIFSS 11 2929
mutmut wildwild
INHINHRR 6767 44 94%94%INHINHSS 88 2211
RIFRIFRR 2929 1616 64%64%RIFRIFSS 00 88
mutmut wildwild
INHINHRR 2424 66 80%80%INHINHSS 11 66
RIFRIFRR 1414 1313 53%53%RIFRIFSS 00 55
mutmut wildwild
INHINHRR 1515 22 88%88%INHINHSS 22 22
RIFRIFRR 1414 22 87.5%87.5%RIFRIFSS 00 55
Харьков- Ukraine Нижний Новгород- RUS
Кемерово- RUS Астана - Kazakhstan
Ser315Thr KatG and Ser531Leu rpoB mutationsSer315Thr KatG and Ser531Leu rpoB mutations
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CodonCodon NovosibirskNovosibirsk KharkivKharkiv
512 Ser->Gly512 Ser->Gly AGC->GGCAGC->GGC 11 --
516 Asp->Val516 Asp->Val GAC->GTCGAC->GTC 77 44
516 Asp->Leu516 Asp->Leu GAC->CTCGAC->CTC -- 11
522 Ser->Leu522 Ser->Leu TCG->TTGTCG->TTG 33 --
526 His->Asp526 His->Asp CAC->GACCAC->GAC 22 --
526 His->Asn526 His->Asn CAC->AACCAC->AAC 11 --
526 His->Tyr526 His->Tyr CAC->TACCAC->TAC -- 44
526 His->Leu526 His->Leu CAC->CTTCAC->CTT -- 11
531 Ser->Leu531 Ser->Leu TCG->TTGTCG->TTG 2121 3333
533 Leu->Pro533 Leu->Pro CTG->CCGCTG->CCG -- 11
513 Gln->term513 Gln->term526 His->Tyr526 His->Tyr
CAA->TAACAA->TAACAC->TACCAC->TAC
-- 11
509 Ser->Thr509 Ser->Thr511 Leu->Pro511 Leu->Pro
AGC ->ACC AGC ->ACC CTG -> CCGCTG -> CCG
-- 11
No mutationNo mutation 55 33
totaltotal 4040 4949
Comparison of rpoB mutations spectrum in Comparison of rpoB mutations spectrum in Novosibirsk and KharkivNovosibirsk and Kharkiv
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Древо филогенетического родства штаммов M. tuberculosis,
полученных из Харькова.
Lin
ka
ge
Dis
tan
ce
0,00
0,05
0,10
0,15
0,20
0,25
0,30
0,35
0,40
12
69
4.m 46
12
21
05
10
2.m 98
78
.m7
6.m 75
74
61
44
.m 41
38
37
36
.m3
5.m 27
24
.m2
3.m 11
7.m 8
41
51
21
25
42
.m 40
12
311
511
0.m
10
91
03
.m1
00
92
.m 86
85
.m8
3.m
71
.m 65
62
.m 57
53
.m 34
17
.m1
0.m 54
10
14
72
61
8 9 811
3.m 31
87
.m 22
12
49
01
9 5 4 73
21 6 3 2
44
5
44
2
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Beijing strains
445
S42 strains
222
West East
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ConclusionsConclusions
S42 (222) is probably endemic for S42 (222) is probably endemic for west territory of prime SU and moves west territory of prime SU and moves to eastto east
This strain possesses different This strain possesses different mechanism of genome evolutionmechanism of genome evolution
Beijing (445) is probably endemic for Beijing (445) is probably endemic for Kazakhstan and acquire the drug Kazakhstan and acquire the drug resistance laterresistance later
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Thank you for attentionThank you for attention