实验动物与比较医学 ›› 2015, Vol. 35 ›› Issue (6): 467-472.DOI: 10.3969/j.issn.1674-5817.2015.06.008

• 论著 • 上一篇    下一篇

猴巨细胞病毒巢式PCR检测方法的建立与初步应用

王莎莎, 贺争鸣   

  1. 中国食品药品检定研究院重点实验室, 北京 100050
  • 收稿日期:2015-04-30 出版日期:2015-12-25 发布日期:2015-12-25
  • 作者简介:王莎莎(1989-), 女, 硕士, 研究方向: 实验动物病毒学。E-mail: wangsha0316@126.com。

Establishment of Nested PCR Detection Method for Simian Cytomegalovirus and Preliminary Application

WANG Sha-sha, HE Zheng-ming   

  1. National Institute for Food and Drug Control, Beijing 100050, China
  • Received:2015-04-30 Online:2015-12-25 Published:2015-12-25

摘要: 目的 建立猴巨细胞病毒(SCMV)特异的巢式PCR检测方法并研究实验猴群和猴源性生 物制品中SCMV感染或污染情况。方法 比对分析多株SCMV序列,设计SCMV特异引物,优化巢式PCR实验条件,建立的巢式PCR方法经验证后检测实验猴群和猴源性生物制品。 结果 经特异度和灵敏度鉴定,在设计的4对引物中确定一对最佳引物,可以区分SCMV和人巨细胞病毒 (HCMV)、小鼠巨细胞病毒(MCMV)、单纯疱疹病毒(HSV)以及犬疱疹病毒(CHV),且可以检测到的 最小DNA量为18 pg/mL。PCR方法检测实验猴群和相关生物制品,发现SCMV在实验猴群中广 泛存在,在相关生物制品中有感染的风险。 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SCMV高度流行,应加强实验猴群及相关生物制品中SCMV的监测, 避免人类感染SCMV的潜在风险。

关键词: 猴巨细胞病毒(SCMV), 巢式 PCR, 实验猴群, 猴相关生物制品

Abstract: Objectives To establish simian cytomegalovirus (SCMV) specific nested PCR detection method, and to investigate SCMV infectious status in monkeys and related biological products. Methods Designed primers based on SCMV sequences and different monkey species and biological products were used to optimize the experiment. Results The established nested PCR detection method for SCMV can distinct SCMV from HCMV, MCMV, HSV and CHV, the test sensitivity is to viral DNA 18 pg/mL. By this nested PCR method, SCMV positive results were detected in monkeys and biological products, which suggested the high infection rate existed in monkeys and its biological products. Conclusions Nested PCR detection method has a good accuracy and sensitivity. It is found that high prevalence in Chinese primate colony. It is necessary to enhance detection for SCMV in monkeys and relevant biological products and to avoid potential risk of infecting in human.

Key words: Simian cytomegalovirus (SCMV), Nested PCR, Detection and application

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