猴腺病毒(SAdV)PCR检测方法的建立及初步应用
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张荣建(1986-),男,免疫学硕士,研究方向:实验动物病毒学。

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国家科技支撑计划课题(No.2011BAI15B01)。


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

    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。

    Abstract:

    ObjectiveTo 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. ResultsA 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. ConclusionsThis 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.

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张荣建,贺争鸣.猴腺病毒(SAdV)PCR检测方法的建立及初步应用[J].中国比较医学杂志,2012,(12):41~47.

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