微生物实验室通则generaloperatingformicrobiologylaboratory

1、微生物实验室通则1.0Purpose/ 目的To define and describe the steps necessary to ensure proper control of the microbiology laboratory, including equipment monitoring, handling of samples, nonconforming product and documentation and review.定义和描述针对合理控制微生物实验室的必要措施，包括设备监测、样品处理、不合格产品、文件及其审核。2.0Scope/ 范围This procedure

2、 applies to all personnel involved directly with the microbiology laboratory and associated support personnel.本程序适用于所有直接涉及微生物实验室的人员以及相关辅助人员。3.0Associated Materials/ 相关材料3.1 QA-LAB-041 Sampling and Testing of the Purified Water SystemQA-LAB-0413.2 QA-LAB-005QA-LAB-0053.3QA-LAB-011QA-LAB-0113.4QA-LAB-

3、023QA-LAB-023纯化水系统取样和检验Handling of Microbiology Laboratory Waste 微生物实验室废弃物处理General Laboratory Procedure实验室通则Laboratory Investigation Procedure实验室调查程序3.5 FM-QA-00024Test ReportFM-QA-00024 检验报告3.6 FM-QA-00025Data SheetFM-QA-00025 原始记录3.7FM-QA-00054FM-QA-00054Micro Test Log 微生物检验登记表3.8FM-QA-00043FM-QA

4、-000433.9FM-QA-000443.10FM-QA-00044FM-QA-00045Instrument Temperature Record设备温度记录Autoclave Performance Check FormLaminar Flow Bench/Biosafty Cabinet Operational Form 超净台/生物安全柜使用记录FM-QA-000453.11FM-QA-00125FM-QA-001253.12FM-QA-00047FM-QA-00047高压灭菌器运行确认记录Instrument Using Log设备使用记录Microbiology Lab Clea

5、ning/Disinfection Form微生物实验室清洁/消毒记录4.0Responsibility/ 职责4.1It is the responsibility of the analyst to ensure that the procedure is followed as written.分析人员的职责是严格按照以下所写程序操作。4.2It is the responsibility of the Microbiology Lab Supervisor to assess any deviations from the written procedure.微生物实验室主管的职责是评

6、估任何来自所写程序的偏差。4.3 It is the responsibility of the Lab Manager to assess the competence of the analyst performing the test.实验室的职责是评估分析人员执行此程序的能力。5.0 Procedure/ 程序5.1 Wearing Apparel 着装5.1.1 Laboratory smocks must be worn whenever any microbiological activity is engaged. When going to the clean room in

7、 microbiology laboratory, must put on the sterile clothes.任何时候进行微生物操作都必须穿实验室工作服。当进入微生物实验室洁净区时，必须要穿上无菌衣物。5.1.2 Non-sterile gloves disinfected with 70% IPA or other suitable disinfectant are to be worn during any testing or handling of cultures. The wearing of sterile gloves is optional.检验或者处理培养基的过程中，

8、必须穿戴消毒过的非无菌手套，可以用70%异丙醇或其他合适的消毒剂消毒。也可选择无菌手套。5.2 Safety 安全5.2.1 All personnel must wash their hands before and after performing any microbiological activity.在进行与微生物有关的工作前后，操作人员必须洗手。5.2.2 Personnel with open sores and rashes must exercise extreme caution while working in the microbiology laboratory.有开

9、放性伤口或者皮疹的人员在微生物实验室工作时必须额外小心。5.2.3 Eating, drinking, smoking and applying cosmetics is strictly prohibited in the microbiology laboratory.微生物实验室内禁止进食、饮水、吸烟和化妆。5.2.4 Aseptic technique must be practiced at all times to prevent contamination of samples, cultures, and sterile culture media. Aseptic techn

10、ique may involve using sterilized (autoclaved) utensils such as forceps or splints, disinfecting metal utensils using 70% IPA and flaming prior to use, using sterile sample cups, working in a laminar flow hood, using an open flame near the weight scale while dispensing samples, wearing sterile or di

11、sinfected garment or gloves, disinfecting all work surfaces, etc.任何时候都要坚持无菌操作，以避免对样品、培养物和无菌培养基介质的污染。无菌操作可包括使用灭菌(经高压灭菌)的器具比如钳子或夹板、使用金属器具前用70% IPA 和火焰进行消毒、使用无菌样品杯、在超净台中操作、分配样品时在天平附近使用明火、穿戴无菌服或消毒手套和消毒工作台面等。5.3 General Guidelines 常规要求5.3.1 Disinfection/Decontamination 消毒/净化5.3.1.1 Treat bench tops and w

12、ork surfaces with 70% isopropyl alcohol before and after each use.用 70% 异丙醇处理使用前后的椅子和工作台面。5.3.1.2 All potentially infectious agents must be disposed of in a disposal bag, details see SOP QA-LAB-005. This includes inoculated media, bacterial cultures, glassware and disposable instruments.具有潜在传染性的物品必须

13、用废弃物处理袋处理，详见SOPQA-LAB-005 。这些包括接种培养基、菌种、玻璃器具和一次性器具。5.3.1.3 Daily, weekly and monthly tasks for disinfection (using 70% IPA or other recognized disinfection agent) of the lab and equipment is to be performed and documented according to FM-QA-00047.执行 FM-QA-00047 的规定并记录实验室及设备的每天、每周和每月的消毒工作(使用70% 异丙醇或其

14、他认可的消毒剂)Daily: Disinfect all lab bench top surfaces, laminar hood work surface and balance prior to use.每日：使用前对实验室的所有台面、超净台工作表面以及天平 进行消毒。Weekly: Autoclave all microbiology laboratory micropipettors in use. Disinfect interior chamber and shelves of all incubators.每周：对微生物实验室使用微量移液枪灭菌消毒。对培养箱的内胆和架子进行消毒。

15、Monthly: Disinfect interior chamber and shelves of refrigerator. Clean water bath and replenish with fresh water.每月：对冰箱的内胆和架子进行消毒。清洁水浴锅，并换水。FM-QA-00047is to be initialed in the appropriate space as tasks are completed. Completed form is to be reviewed.完成任务后，应在FM-QA-00047 中记录，完成的记录表应被复核。设备监测和操作5.3.2

16、Equipment Monitoring and Operation5.3.2.1Record the equipments temperatures on the appropriate forms,or record the running status on the appropriate forms:将设备的温度并记录在对应的监测记录上，或者将设备的运行状态记录在相应的记录上：Laminar flow hood and Biosafety cabinet超净台和生物安全柜台FM-QA-00043Incubator 培养箱FM-QA-00044Incubator temperature

17、recorder 培养箱温度记录仪FM-QA-00044Refrigerator 冰箱 Centrifuge 离心机 Stomacher 匀质器 Autoclave 灭菌锅 Isolator 隔离器FM-QA-00105(seeFM-QA-00125(seeFM-QA-00125(seeFM-QA-00125(seeFM-QA-00125(see见 QA-LAB-079)见 SOP-LAB-011)见 SOP-LAB-011)见 SOP-LAB-011)见 SOP-LAB-011)For the Micro Fridge, also record the Calibration Due Dat

18、e from the immersed thermometer.对于微生物冰箱，还需要记录内置温度计的校验到期日。5.3.2.2 If an out of limits condition is detected, the Laboratory Supervision will be notified immediately. Corrective action will be as follows:如果发现了超限情况，立即通知实验室主管。纠正措施如下：INCUBATORS 培养箱- - If the temperature is above the specified range, all

19、material within will be purged and product in-test will be re-tested. 如果温度高于规定范围，那么其中的物品都应废弃，且检验中的产品应重新检验。- - If the temperature is below the specified range, all items within will be incubated one extra day for each day temperature was out of limits.如果温度低于规定范围一天，那其中的培养物都需多培养一天。- - If the non-confor

20、mance does not self-correct, the temperature set point may be adjusted followed by a requalification (temperature mapping) to ensure that the set point change does not push certain locations inside the chamber out of temperature range.如果无法自行纠正，那么需要重新确认（温度分布）后，调整温度设定值，以确保设定值的变化不会使得内胆中局部位置的温度超出范围。REFR

21、IGERATORS 冰箱- - Immersed thermometer will also be read daily to determine the actual product temperature as compared to the ambient air temperature within the fridge.应每天读取内置式温度计以确定产品实际的温度，相对于冰箱内部的空气温度。- - If the temperature is above the specified range, then the microbiological media will be inspect

22、ed for contamination or unusual appearance. Any articles that present contamination or changes from normal appearance will be discarded. All other items will be retained.如果温度高于规定范围，应检查微生物培养基，是否有污染或异常外观。应销毁有污染或者异常外观的，保留其他物品。- - If the temperature is below the specified range, check supplies for signs

23、 of freezing. Discard any frozen media. If temperature is below range for more than 24 hours all material must be discarded.如温度低于规定范围，应检查所有物品是否有过冷迹象。销毁所有已冻结培养基。如温度低于规定范围超过24 小时，应销毁所有的物品。- - If the non-conformance does not self-correct, the temperature set point may be adjusted using the manual dial.

24、 The adjustment should be made based on the temperature value using an immersed thermometer at the worst case location within the fridge. An adjustment within this range will ensure that the set point change does not push certain locations inside the chamber out of temperature range.如果无法自行恢复正常，可以利用手

25、动调节设置温度。调节时应按照冰箱内条件最差方位的温度计读数进行，按此温度进行调整可以确保变更设定值不会使局部温度超限。LAMINAR FLOW HOOD and BIOSAFETY CABINET超净台和生物安全柜- - When the instrument can not use, the cabinet should be re-certified by a qualified technician. Hood may be used for testing purposes prior to re-certification if particle count check as per

26、 5.3.3.1 is performed daily with passing results.当设备出现问题，则要请有资质的技术人员重新认证。如果按照 5.3.3.1 执行每日检查且结果为合格，那么对超净台进行重新认证以前可以将其用于检验。CENTRIFUGE 离心机Calibrate the centrifuge every month .if the result exceed the warning limit ,repeat check calibration, If warning limit still exceeded ,inform section leader who w

27、ill investigate the problem. If the result exceeds the action limit, repeat check calibration, If action limits still exceeded, affix notice indicating, timer is withdrawn from use. Inform Section Leader to investigate problem.每月校验离心机。如果结果超出警戒限，重复检查校验，如果限值仍然超出，告知部门主管，由他来调查问题。如果结果超出行动限，重复检查校验。如果行动限仍然

28、超出，应贴上通知说明计时器不再使用。通知主管调查问题。STOMACHER 匀质器Calibrate the stomacher every month .if the result exceed the warning limit ,repeat check calibration, If warning limit still exceeded ,inform section leader who will investigate the problem. If the result exceed the action limit, repeat check calibration, If

29、action limits still exceeded, affix notice indicating ,timer is withdrawn from use. Inform Section Leader to investigate problem.每月校验匀质器。如果结果超出警戒限，重复检查校验，如果限值仍然超出，告知部门主管，由他来调查问题。如果结果超出行动限，重复检查校验。如果行动限仍然超出，应 贴上通知说明计时器不再使用。通知主管调查问题。Nonconformities, corrective action and impact on test samples/micro me

30、dia may be documented on the applicable forms or charts. The general lab OOS procedure does not need to be followed in these instances. 在相应的表格或者图表上记录不合格、纠正措施和对检验样本 /微生物培养基的影响。在此情况下，无需执行实验室OOS 。5.3.3 Monitoring of the Laminar Flow Hood, Autoclave and Isolator 超净台、高压灭菌器和隔离器的监测5.3.3.1 To the Laminar Fl

31、ow Hood, If at any time during the monitoring, growth is exhibited on any of the tryptic soy agar plates used in the monitoring process (when doing the sterility testing for china market, it is necessary to monitor the Laminar Flow Hood, clean room and the operators hands with two TSAplates respecti

32、vely ) or particles are registered when5.3.3.25.3.3.35.3.3.4performing particle counts, the Laminar Flow Hood must be requalified for five (5) consecutive days by repeating the above test. Hood may be used at risk pending passing results.However, if failing results are obtained on any of the five da

33、ys, any failing microbiological tests conducted within the hood during that time period may be nullified.对于超净台，如果在监测中，TSA 培养皿上出现了培养物(检测中国市场无菌产品时，实验同时需要分别用2 块营养琼脂平板进行超净台、洁净室和操作人员手的监测)，或在进行悬浮粒子计数时记录下颗粒物，那么应对超净台进行连续五天的重新确认。在等待结果期间，超净台可以使用，但有风险。但是，如果在五天中任一天得到不合格结果，那么那段时间内在超净台内所进行的任何不合格微生物检验都可能无效。The aut

34、oclave must be monitored quarterly to ensure the continued effectiveness of the sterilization process. Monitoring will consist of three (3) BIs (spore strips of Bacillus stearothermophilus , population 10 6/strip). One BI will be placed in the top of the chamber, one in the centre and one in the sub

35、strate. A validated liquids cycle will then be initiated.After the cycle is complete, the BIs will be incubated at 55 oC - 60 o C for a minimum of seven (7) days. BI will also be incubated. If, at any time during the monitoring, growth is exhibited from sterilized BI used in the monitoring process,

36、then a qualified technician is to be called in to inspect and adjust the autoclave as necessary. Then the autoclave must be re-qualified with the BI test for consecutive three times.必须每三个月对高压灭菌器进行监测，以确保灭菌工艺的持续有效性。监测由三(3)个BI (带嗜热脂肪芽抱杆菌，浓度10 6/条)组成。灭菌器上部、中部和下部各放置一个BI 。随后开始已验证的液体程序。程序完成后，BI 应在 55oC - 6

37、0 o C 的温度下至少培养7 天。还应该对BI 进行培养。如果在培养中发现有生长，那应请有资质的技师进行检查并对高压灭菌器进行必要的调节。随后，必须用BI 对高压灭菌器进行连续三次的检验进行重新确认。In normal testing, the isolator needs to change the gloves on the first working day, then run the Decay test and Decon cycle later. (details see SOP QA-LAB-094)当需要进行无菌检验时，在每周第一个工作日进行手套更换，完成后进行Decay 泄

38、露测试，继而进行Decon 灭菌。(具体操作见 SOP QA-LAB-094 )All documentation concerning each month or each time monitoring of both the Laminar Flow Hood , autoclave(s) and the isolator must be recorded on the applicable check forms and filed within Quality Assurance. Any nonconformities, corrective action and impact o

39、n test samples/micro media will be documented on the applicable form. The general lab OOS process does not need to be followed in these instances. 所有与超净台、高压灭菌器和隔离器每月或每次监测有关的文件都必须记录在相应的检查表格上，并由QA 部门归档。应在合适的表格上记录不合格、纠正措施以及对检验样品/微生物培养基的影响。在此情况下，不需要遵守实验室OOS 。5.3.4 Media, Reagents and standard culture 培养

40、基、试剂和标准菌种5.3.4.1 All laboratory prepared media and reagents must be Quality Control tested prior to use for sterility and performance. All commercial media must be examined. All standard culture and microbial suspensions are to be recorded in the check in log. See applicable SOP for proper methods,

41、documentation, and laboratory check-in procedures.所有实验室制备的培养基和试剂在进行无菌使用前都必须进行质量控制检验，检查所有的商用培养基和所有标准菌种，并记录在检查表上。适用的方法、文件记录以及实验室登记程序参阅相应的SOP 。5.3.5 Evaluation of Samples 样品评估5.3.5.1 All samples received into the microbiology laboratory should be considered potentially infectious and handled in the app

42、ropriate manner. In all cases, aseptic technique must be practiced to prevent contamination during manipulation of samples, cultures and sterile culture media.微生物实验室接收的所有样品都应视为有潜在传染性，需要合理方式进行处理。在任何情形下都必须采用无菌操作处理样品、菌种和无菌培养基。5.3.5.2 When a sample is received into the microbiology laboratory for testin

43、g, be it a raw material, a developmental product, or finished product, the product name, lot number, and any other pertinent information must be recorded on the applicable Microbiology Data Form and entered into the Micro Test Log FM-QA-00054 (QA-LAB-027). All samples should be accompanied by an Tes

44、t Request.实验室接收检验样品时，不管样品是原料、研发产品或者成品，都必须在相应的微生物原始记录上记录产品名、批号和其他的相关信息，并在微生物检验登记表FM-QA-00054 上进行登记(QA-LAB-027 )。所有的样品都必须附上检验申请表。5.3.5.3 Depending on the sample type, the sample must then be evaluated accordingly per the appropriate material specification. Any test media which is used in the evaluatio

45、n procedure must be labelled with lot number of sample and date in test. Any media which is used in the evaluation process must be documented according to the applicable test procedure. Documentation must include media name, manufacturer, lot number and expiration date. All raw data must be recorded

46、 on the Microbiology Data Form (if applicable) and final results and disposition are to be recorded on the Test Report Form or Test Request.基于样品类型，根据相应的物料标准评估样品。评估过程中所用到的检验培养基都必须标明样品批号和检验日期。评估过程中须根据相应的检验程序记录所使用到的任何培养基。文件记录必须包括培养基名称、厂商、批号和有效期。所有原始数据都必须记录在微生物原始记录(如适用)上，处理方式必须记录在检验报告或者检验申请表上。5.3.5.4 Du

47、ring evaluation, approved component specifications and approved methods of analysis must be referenced at all times. 在评估过程中，须随时参考已批准的成分标准和分析方法。5.3.5.5 Acceptability of the test results of a sample are to be indicated on the Test Report if applicable. Based on the final test results and comparison wi

48、th applicable standards, test limits, or product specifications, the sample will be deemed acceptable or rejectable.有时，要在检验报告上指出样品检验结果的可接受性。根据最终检验结果以及与相应标准、检验限度或者产品标准的对比，确定样品可以接受或者拒绝。5.3.6 Nonconformities 不合格5.3.6.1 Any material found to be non-conforming to approved specifications must be reported

49、immediately to laboratory supervision or approved delegate.发现与已批准标准不相符的物料必须立即报告实验室主管或者其指定的代表。5.3.6.2 OOS results are to be handled by following SOP QA-LAB-023“ Laboratory Investigation Procedure.” Water analysisnonconforming results will be addressed as per SOP QA-LAB-041.按照 SOP QA-LAB-023 “实验室调查程序

50、”处理 OOS 结果。根据 SOP QA-LAB-041 来解释水分析不合格结果。5.3.7 Documentation and Review 文件和审核5.3.7.1 Upon completion of analysis, all pertinent documentation will be compiled by the analyst.完成分析后，由分析师对所有相应文件进行整合。5.3.7.2 The completed documents may be reviewed by a second person who is trained to the pertinent SOPs governing the documents. 可由另一人检查完成的文件，该人接受过管理文件的SOP 培训。5.3.7.3 All final completed Test Report will be forwarded to Quality Assurance to be included in the applicable product history file. Microbiology Data Forms, which contain raw data, Micro Test Log sheets and Microbial Limit