Enzymatic amplification of hepatitis B virus DNA in serum compared with infectivity testing in chimpanzees.
Enzymatic amplification of hepatitis B virus DNA in serum compared with infectivity testing in chimpanzees.
复制标题
血清中乙型肝炎病毒 DNA 的酶扩增与黑猩猩感染性测试的比较。
DOI：
10.1093/infdis/160.1.37
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Vyas,GN
中科院分区：
文献类型：
--
作者：
Ulrich,PP;Bhat,RA;Seto,B;Mack,D;Sninsky,J;Vyas,GN
乙型肝炎病毒（HBV）的体内感染滴定已经按照黑猩猩感染剂量（CID50）进行了生物学标准化。采用聚合酶链反应（PCR）体外酶扩增HBV DNA，并与CID50进行比较。对含有HBVadw（CID5010−7）和ayw（CID5010−7,5）的人血清标本进行系列稀释，以确定PCR检测HBV DNA的重复性和敏感性。无论是从血清中提取的HBV DNA扩增，还是在蛋白水解血清中直接扩增HBV DNA， PCR对血清中HBV DNA的检出限均为10−8。使用0.25%浓度（vol/vol）的非离子型洗涤剂和2.5%浓度（vol/vol）的消化人血清，对蛋白水解物的PCR扩增效率没有显著降低。因此，PCR是一种特异性和快速的体外检测HBV DNA的方法，比黑猩猩接种HBV的体内感染性滴定法更敏感。因此，如果在标本或生物制品中无法通过PCR检测到HBV相关DNA，则它们可能不含有感染性HBV。
The in vivo infectivity titration of hepatitis B virus (HBV) has been biologically standardized in terms of chimpanzee infectious dose (CID50). The polymerase chain reaction (PCR) was used for in vitro enzymatic amplification of HBV DNA and comparison with CID50. Serial dilutions of human serum specimens containing HBVadw(CID5010−7) andayw(CID5010−7,5) were tested to determine the reproducibility and sensitivity of PCR for the detection of HBV DNA. The detection limit of HBV DNA in serum using PCR was 10−8for either amplification of HBV DNA extracted from serum or direct amplification of HBV DNA in proteolyzed serum. The amplification efficiency of PCR in proteolysates was not significantly reduced by using a 0.25% concentration (vol/vol) of non-ionic detergents and 2.5% concentration (vol/vol) of digested human serum. Thus, PCR is a specific and rapid method for in vitro detection of HBV DNA that is more sensitive than the in vivo infectivity titration of HBV by chimpanzee inoculation. Therefore, if HBV-related DNA cannot be detected by PCR in specimens or biologic products, they probably do not contain infectious HBV.