猴腺病毒（SAdV）PCR检测方法的建立及初步应用

目的建立SAdV特异的PCR检测方法并研究实验猴群和猴源性生物制品中SAdV感染或污染情况。方法比对分析多株SAdV序列,设计SAdV特异引物,优化PCR实验条件,建立的PCR方法经验证后检测实验猴群和猴源性生物制品,阳性产物测序并构建进化树。结果经特异性和敏感性鉴定,在设计的5对引物中确定一对最佳引物,可以区分SAdV和MAD,ICH,CELO且可以检测到的最小DNA量为47.9 pg/mL。PCR方法检测实验猴群,阳性率49.2%,测序及进化分析表明,SAdV感染型别呈广泛基因多样性。主要分布在G亚属和以SAdV-49为代表的分支。结论经测序验证,PCR检测方法具有很好准确性,初步应用表明我国实验猴群中SAdV高度流行,应加强实验猴群及相关生物制品中SAdV的监测,避免人类感染SAdV的潜在风险。

Establishment and Preliminary Application of a PCR Detection Method for Simian Adenovirus

Objective To establish a PCR detection method for simian adenovirus（SAdV） and study the SAdV infection status in monkeys.Methods To design a primer based on SAdV sequences and optimize the PCR experiment,test the method,to detect the virus in monkeys and vaccines,and sequenceing and constructing the phylogeny trees.Results A PCR detection method for SAdV was established,which could distinguish SAdV from MAD,ICH,CELO and was sensitive to 47.9 pg/mL viral DNA.The results of this study showed a positive rate of 49.2% and a large genetic diversity of simian adenoviruses in Chinese primate colony.Most of them belong to G subgroup and twig of SAdV-49.Conclusions This PCR detection method established in our laboratory has a good accuracy.We found a high prevalence of SAdV in Chinese primate colony.It is necessary to enhance detection and control for SAdV in monkeys and relevant biological products,and avoid potential risk of human infection.