一批逆转录酶活性检测假阳性的鉴别  

Distinguish the RTase activity and false-positive reactions

作　　者：孔艳[1] 杨立宏[1] 安祺[1] 董关木[1] 俞永新[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《微生物学免疫学进展》2010年第4期26-28,共3页Progress In Microbiology and Immunology

摘　　要：目前逆转录病毒的检测方法被普遍应用的是逆转录酶活性检测,自从PCR方法被应用到逆转录酶活性检测中后,使检测的灵敏度及特异性得到极大地提高。同时,假阳性结果也带给各实验室很大困扰,包括试验过程中的实验用试剂及检测样品本身带来的假阳性。实验中观察了一批逆转录酶活性检测阳性样品和两个依赖DNA的DNA聚合酶的假阳性,通过对检测的严密监控以及改变反应体系pH值,抑制了由于细胞死亡后所释放的细胞内DNA聚合酶以及实验用DNA聚合酶的类逆转录酶样作用,从而达到鉴别结果的真实性的目的。Safety testing is an important issue in detection of retroviruses in cell substrates used to produce biologics and vaccines.Recently,several polymerase chain reaction（PCR） based RT detection assays have been developed,which are more sensitive than conventional nucleotide incorporation assays.The ultra-sensitive product-enhanced reverse transcriptase（PERT） assay has been used to detect minute reverse transcriptase（RTase）.Reverse transcriptase（RT） assays can be used to detect the presence of retroviruses.However,concurrence with this increase in sensitivity is an increment in false-positive reactions from the contamination in experiment and from the DNA dependent DNA polymerase which are known to have RTase-like activities.We have developed a one-step PERT assay with reducing pH described here by using the MS2 RNA template and primers which may significantly reduce the level of false-positive reactions in a more sensitive and specific way.

关 键 词：生物制品 逆转录酶活性检测 假阳性 

分 类 号：R341[医药卫生—基础医学]