wfhss and jsmi conference 2012 - session 8: a study on … · 2014-12-29 · how do we assess...

Annual WFHSS and JSMI Conference 2012 13th World Sterilization Congress

Takayuki Ohishi 1,2), Hiroyoshi Kobayashi 2), Takashi Okubo 2) , Yoshimitsu Hiejima 2) , Ryo Fushimi 2,3), Satoshi Shinomiya 2,4), Eiko Miura 5), Hiroshi Matsumoto 1)

A Study on the Cleaning of Flexible Endoscope

by Adenosine Triphosphate Measure

1) Department of Infection Control and Prevention, Osaki Citizen Hospital, Japan 2) Division of Infection Prevention and Control, Postgraduate School Tokyo Healthcare University, Japan 3) Department of Central Supply and Sterilization Department, Osaka University Hospital, Japan 4) Department of Infection Control and Prevention, Mino City Hospital, Japan 5) Department of Clinical Laboratory, Osaki Citizen Hospital, Japan

AIMS

• 軟性内視鏡の清浄度評価は、洗浄・消毒（再処理）の質を保証するうえで重要

• 清浄度の指標を細菌培養とするのは困難*

• Cleaning evaluation of flexible endoscopes after clinical use is essential for ensuring the quality of reprocessing.

• Bacterial culture is not a suitable indicator*.

* Dancer SJ. How do we assess hospital cleaning? A proposal for microbiological standards for surface hygiene in hospitals. J. Hosp. Infect. 2004;56:10-5.

AIMS • アデノシン三リン酸(Adenosine triphosphate ; ATP)測定による清浄度評価は迅速で有用*

• 再処理前後の鉗子チャンネル内のATPを2試薬で計測し、有効性を検討

• Cleaning evaluation is useful in the rapid by Adenosine triphosphate (ATP) *.

• ATP fluid in endoscopes channel before and after reprocessing was determined by two types of reagents, and efficacy was investigated.

* Hansen D, Benner D, Hilgenhöner M, Leisebein T, Brauksiepe A, Popp W. ATP measurement as method to monitor the quality of reprocessing flexible endoscopes. Ger Med Sci. 2004 Apr 26;2:Doc04.

Principle of ATP detection

* 羽毛田靖: ATP法による細菌数測定装置の基礎と応用. 防菌防黴 25:457-466, 1997. 改変

Sample

bacterium

ATP

ATP

ATP

Extraction reagent

ATP

ATP

ATP

Luminescence reagent

ATP +

Luminescence reagent

Light

Light

ATP + D-luciferin + O2 AMP + oxyluciferin + ppi + Light Luciferase

Mg2+

Extraction

Luminescence

Measurement

Unit is RLU

AIMS (Standardized ATP)

• 主なATP測定試薬の測定差を比較

• ATP標準液を測定し、試薬間の相関を検証

• Compare the difference of measured values by a variety of major ATP reagents.

• Verify the correlation between reagents by measuring standardized ATP solution.

SAMPLE COLLECTION METHODS




検体採取法（前液） Sample collection (1st-fluid)

① 患者から内視鏡抜去 ② 10mLの滅菌蒸留水を鉗子口金から急

速に注入 ③ 先端から抽出液を滅菌スピッツに採取

（前液） Forceps channel

Sterile tube

Syringe

Flexible endoscopes

1. After clinical use of flexible endoscopes

2. Sterile distilled water (10mL) was injected into the forceps channel

3. The fluid (1st-fluid) was collected into sterile tube

Reprocessing of flexible endoscopes ① 内視鏡先端から水道水→酵素洗浄剤溶液→

水道水の順で吸引

Tap water Enzyme detergent Tap water

1. Suction Tap water→Enzyme detergent fluid → Tap water

Reprocessing of flexible endoscopes

② チャンネル内を三方向からブラッシング

Forceps channel

Suction cylinder

Suction channel

2. Brushing from three directions in the channel


③ 吸引口金より吸引器でチャンネル内を洗浄

Suction unit

Suction channel

3. Cleaning the channel by suction unit


④ 自動内視鏡洗浄装置にて洗浄・消毒

Automatic endoscope reprocessor

4. Cleaning and disinfection by automatic endoscope reprocessor

検体採取法（後液） Sample collection (2nd-fluid)

①10mLの滅菌蒸留水を鉗子口金から急速に注入

②先端から抽出液を滅菌スピッツ

に採取（後液）

1. Sterile distilled water was injected into the forceps channel

2. The fluid (2nd-fluid) was collected into sterile tube

Forceps channel

Sterile tube

Syringe

Flexible endoscopes

Sample state

• 前液と後液はそれぞれ20検体を採取 • 採取は1日で終了

• 検体採取後、直ちに各種測定を実施

• 20 samples of 1st-fluid and 2nd-fluid were collected respectively in one day.

• Samples were measured immediately by a variety of testing methods.

MEASUREMENT INSTRUMENT & REAGENT




Measurement methods • 前液と後液をATP測定機器と試薬で、1検体あたり1回、規定量を測定

• 検体は試薬にチャージ後、30秒以内に測定 • 生菌数確認の細菌培養を実施

• Defined volume of 1st-fluid and 2nd-fluid were measured once per sample by ATP measuring instrument and reagent.

• After charging the reagent, the sample was measured within 30 seconds.

• Bacterial cultures were conducted to confirm the living bacteria.

A-test B-test

Instrument

LumitesterTM PD-20 (PD)

CleanTraceTM UNG3 (UNG)

Reagent LuciPacTM Pen (LPP)

Clean TraceTM AQT (AQT)

Marketer Kikkoman 3M

ATP measuring instrument & reagent

Shape and collection volume of each reagents

Reagent

Tip

Volume

(μL)

Picture

A-test LPP Swab 115

B-test AQT Groove 160

1.4 times

Bacterial culture

ヒツジ血液寒天培地 24時間好気培養（定量）

blood agar and cultured for 24 hours at 37℃

STANDARDIZED ATP SOLUTION MEASUREMENT




Standardized ATP solution

• ATP （Adenosine-5‘-triphosphate：オリエンタル酵母社）を滅菌蒸留水に溶解

• 10-12 ～10-7 mol/Lの希釈系列を作成

• The ATP (Adenosine-5‘-triphosphate： Oriental Yeast Co., ltd.) was dissolved into sterile distilled water.

• Serial dilution was performed to prepare 10-12 ～10-7 mol/L.

Standardized ATP solution measurement

• ATP測定はPDとLPP（A-test）、UNGとAQT（B-test）で実施

• 測定量は規定量とし、各々30回ずつ測定

• Standardized ATP solutions were measured by PD and LPP (A-test), UNG and AQT (B-test).

• Defined volume were measured 30 times respectively.

Presenter

Presentation Notes

(s2) Amd → and (s3)30 times→10times

RESULTS




ATP and bacterial culture A-test (RLU) B-test (RLU) Bacterial

Culture (CFU)

1st-fulid 2nd-fulid 1st-fulid 2nd-fulid 1st-fulid 2nd-fulid

Median 646,485 26.8 476,908 49.5 1065 0

25~75 Percentile

624,200 62~

625,253

24.8 2~

28.0

476,633 476,~

428,632

31.5 ~

76.3

82.5 ~

1065 0

Maximum 199,513 22.8 470,042 146.5 1065 0

Minimum 682,036 22.8 476,355 122.5 05 0

n=20

1

10

100

1,000

10,000

100,000

1,000,000

1st fluid 2nd fluid 1st fluid 2nd fluid 1st fluid 2nd fluid

A-test A-test Microbiologicalcultures

ATP

mea

sure

men

ts (

RLU

) an

d nu

mbe

r of

org

anis

ms

(CFU

/mL)

ATP and bacterial culture

＊t - test

p = 0.022* p = 0.068* p < 0.001*

B- test

Correlation of two types of reagents

y = 0.6561x + 1.1732 R = 0.294 p = 0.066*

0

1

2

3

4

5

6

0 1 2 3 4 5 6

B-te

st (

RLU

)

A-test (RLU) ＊t - test

log

log

n=20

STANDARDIZED ATP SOLUTION RESULTS




Measurement of standardized ATP

ATP (mol/L)

AV (log) SD(log) CV(%) AQT LPP AQT LPP AQT LPP

10-12 2.50 1.68 0.08 0.07 3.15 4.14 10-11 2.83 1.94 0.07 0.04 2.37 2.09 10-10 3.55 2.61 0.09 0.03 2.45 1.02 10-9 4.52 3.51 0.07 0.04 1.63 1.17 10-8 5.36 4.43 0.10 0.04 1.90 1.02 10-7 6.03 5.38 0.03 0.05 0.50 0.87 RO

water 1.35 0.69 0.11 0.18 8.20 26.58

n=30

Correlation of A-test and B-test

y = 0.9674x + 0.979 R=0.99661

p = <0.0001*

0

1

2

3

4

5

6

7

0 1 2 3 4 5 6

B-te

st (

RLU

)

A-test (RLU) log

n=30 log

＊t - test

DISCUSSIONS




Discussions

• A-test、B-test共に再処理後のRLUが激減し、

細菌培養結果と比例したため、軟性内視鏡の

清浄度評価としてATP測定の有用性が示

唆

• The results indicate that ATP is clearly beneficial for assessing the efficacy of cleaning and disinfection of reprocessing flexible endoscopes.

Presenter

Presentation Notes

(s2)beneficial in → beneficial for

Discussions

• 抽出液測定における2試薬の相関が弱かったの

は、A-testがATPと共にアデノシン一リン酸も測定することや、抽出液中の上皮組織などが

測定誤差を誘発している可能性

• The low correlation between the results of the two test methods may be attributed to the inclusion of adenosine monophosphate in A-test, epithelial tissues in the solution, and to the difference in the conversion factors.

Discussions

• ATP標準液測定による2試薬間の相関は非常に

強い（R=0.99661）が、 A-testの測定結果はB-testより1log程度低くなることが判明

• The strong correlation (R=0.99661), standardized ATP solution measurements indicated that the A-test is 1 logs lower than the B-test.

Conclusions

• ATP測定は、簡便、迅速、定量的であり、再処

理が必要な軟性内視鏡の清浄度評価として有効な方法

• 内視鏡を再処理する目安となるATPの目標値や

許容値設定の検討が必要 • ATP measurement that is rapid, convenient,

quantitative is an effective methods for evaluation of reprocessing flexible endoscopes.

• We must establish a reference value for reprocessing.

wfhss and jsmi conference 2012 - session 8: a study on … · 2014-12-29 · how do we assess...

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