鸭疱疹病毒1型套式PCR检测方法的建立及应用

为建立一种快速鸭疱疹病毒1型(Anatid herpesvirus 1,AHV-1),又名鸭肠炎病毒(duck enteritis virus,DEV)病原检测方法,本研究根据GenBank上登录的DEV基因序列,设计合成内、外2对引物,建立了检测DEV的套式PCR方法。该方法对正常鸭胚、健康鸭肝肠组织、鸡传染性喉气管炎病毒、伪狂犬病病毒、减蛋综合征病毒、鸭肝炎病毒和新城疫病毒的扩增结果均为阴性;该方法第1次扩增的敏感性是10 ng,第2次扩增的敏感性是0.1 ng,第2次比第1次扩增的敏感性高100倍。建立的套式PCR方法具有良好的特异性、敏感性,可以准确快速检测出极低含量的DEV,将为鸭病毒性肠炎的病原检测及分子流行病学调查等提供一种高效、快速、特异、灵敏的检测方法。

Establishment and Application on a Nest PCR Method for Detection of Anatid Herpesvirus 1

To establish a rapid duck enteritis virus (DEV) pathogen detection method, according to the DEV gene sequence in GenBank, we synthesized outer and inner two pairs of primers, established a nested PCR for DEV detection. The method used for normal duck embryo, healthy duck constitution, infectious laryngotracheitis virus (ILTV), pseudorabies virus (PRV), egg drop syndrome virus (EDSV), duck hepatitis virus (DHV) and Newcastle disease virus (NDV), all of the PCR results were negative, the sensitivity of first amplification was 10 ng, the second amplification sensitivity was 0.1 ng, the second amplification was more sensitive 100 times than the first amplification. The established nest PCR method had good specificity, sensitivity, it could detect low levels of DEV quickly and accurately, it provided a kind of efficient, rapid, specific and sensitive detection method for pathogen detection and molecular epidemiology of material such as of duck viral enteritis.