国际生物制品学杂志
國際生物製品學雜誌
국제생물제품학잡지
INTERNATIONAL JOURNAL OF BIOLOGICALS
2015年
2期
63-66
,共4页
许敬敏%吕冰凌%刘菊%潘海龙
許敬敏%呂冰凌%劉菊%潘海龍
허경민%려빙릉%류국%반해룡
聚合酶链反应%分析仪器确认%药品生产质量管理规范
聚閤酶鏈反應%分析儀器確認%藥品生產質量管理規範
취합매련반응%분석의기학인%약품생산질량관리규범
Polymerase chain reaction%Analytical instrument qualification%Good manufacture practice
目的 通过对实时荧光定量PCR(real-time fluorescent quantitative PCR,RTFQ-PCR)仪实施确认,了解分析仪器确认的具体步骤.方法 根据药品生产质量管理规范的要求,对该RTFQ-PCR仪分别进行安装确认、运行确认和性能确认,以及年度再确认.在性能确认中,通过定量PCR法和环介导恒温(loop-mediated isothermal amplification,LAMP)法检测分枝杆菌核酸来验证该仪器的准确性和重复性.结果 RTFQ-PCR仪的安装符合要求,各项测试操作均达到规定的标准.当采用定量PCR法检测TB进行RTFQ-PCR仪性能确认时,结核杆菌(tubercle bacillus,TB)核酸浓度的对数与循环阈值(cycle threshold,Ct)之间呈现良好的线性关系,相关系数绝对值达0.999 9;TB核酸回收率为97％～105％;对2个TB样品分别进行3次检测获得的Ct值的变异系数(C聊均小于5％.当采用LAMP法检测分枝杆菌进行RTFQ-PCR仪性能确认和再确认时,对分枝杆菌阳性对照和样品分别进行3次检测获得的Ct值的CV均小于5％.结论 该RTFQ-PCR仪得到成功确认,各方面均符合规定的要求.
目的 通過對實時熒光定量PCR(real-time fluorescent quantitative PCR,RTFQ-PCR)儀實施確認,瞭解分析儀器確認的具體步驟.方法 根據藥品生產質量管理規範的要求,對該RTFQ-PCR儀分彆進行安裝確認、運行確認和性能確認,以及年度再確認.在性能確認中,通過定量PCR法和環介導恆溫(loop-mediated isothermal amplification,LAMP)法檢測分枝桿菌覈痠來驗證該儀器的準確性和重複性.結果 RTFQ-PCR儀的安裝符閤要求,各項測試操作均達到規定的標準.噹採用定量PCR法檢測TB進行RTFQ-PCR儀性能確認時,結覈桿菌(tubercle bacillus,TB)覈痠濃度的對數與循環閾值(cycle threshold,Ct)之間呈現良好的線性關繫,相關繫數絕對值達0.999 9;TB覈痠迴收率為97％～105％;對2箇TB樣品分彆進行3次檢測穫得的Ct值的變異繫數(C聊均小于5％.噹採用LAMP法檢測分枝桿菌進行RTFQ-PCR儀性能確認和再確認時,對分枝桿菌暘性對照和樣品分彆進行3次檢測穫得的Ct值的CV均小于5％.結論 該RTFQ-PCR儀得到成功確認,各方麵均符閤規定的要求.
목적 통과대실시형광정량PCR(real-time fluorescent quantitative PCR,RTFQ-PCR)의실시학인,료해분석의기학인적구체보취.방법 근거약품생산질량관리규범적요구,대해RTFQ-PCR의분별진행안장학인、운행학인화성능학인,이급년도재학인.재성능학인중,통과정량PCR법화배개도항온(loop-mediated isothermal amplification,LAMP)법검측분지간균핵산래험증해의기적준학성화중복성.결과 RTFQ-PCR의적안장부합요구,각항측시조작균체도규정적표준.당채용정량PCR법검측TB진행RTFQ-PCR의성능학인시,결핵간균(tubercle bacillus,TB)핵산농도적대수여순배역치(cycle threshold,Ct)지간정현량호적선성관계,상관계수절대치체0.999 9;TB핵산회수솔위97％～105％;대2개TB양품분별진행3차검측획득적Ct치적변이계수(C료균소우5％.당채용LAMP법검측분지간균진행RTFQ-PCR의성능학인화재학인시,대분지간균양성대조화양품분별진행3차검측획득적Ct치적CV균소우5％.결론 해RTFQ-PCR의득도성공학인,각방면균부합규정적요구.
Objective To understand the specific contents of analytical instrument qualification by implementing qualification of the real-time fluorescent quantitative PCR (RTFQ-PCR) system.Methods According to the requirement of GMP,installation qualification,operational qualification,performance qualification (PQ) and requalification of the RTFQ-PCR system were implemented.In PQ,accuracy and repeatability of the RTFQ-PCR system were validated by detecting mycobacteria DNA with quantitative PCR method and loop-mediated isothermal amplification (LAMP) method.Results The RTFQ-PCR system installation met the requirement,and each testing operation reached the required standard.When PQ of the RTFQ-PCR system was implemented by quantitative PCR detection,threre was a good linear relationship between the logarithm of tubercle bacillus (TB) DNA concentration and cycle threshold (Ct),and the absolute value of correlation coefficient reached 0.999 9.Recoveries of TB DNA were 97％-105％.Coefficient variations (CVs) of Cts obtained by detecting two TB samples for three times each with quantitative PCR method were both less than 5％.When PQ and requalification of the RTFQ-PCR system were implemented by LAMP detection,CVs of Cts obtained by detecting the positive control and the sample of mycobacteria for three times each with LAMP method were all less than 5 ％.Conclusion Qualification of the RTFQ-PCR system is implemented successfully,and all aspects of the RTFQ-PCR system meet requirement of analytical instrument qualification.
