番鸭三种细小病毒多联PCR检测方法的建立与初步应用

Establishment and Preliminary Application of Multi-PCR for Diagnosis of Three Parvovirus in Muscovy Ducks

根据GenBank中番鸭"三周病"病毒(MDPV)、小鹅瘟病毒(GPV)的NS基因和VP基因,以及华南农业大学禽病组分离到的细小病毒型"白点病"病毒(MDWNV)的NS和VP基因序列,设计3对特异性引物,并在建立单项PCR技术的基础上,优化多联PCR反应条件(EX-Taq DNA聚合酶浓度、引物浓度、dNTPs浓度以及退火温度等),建立了MDPV、MDWNV和GPV三种病毒的多联PCR检测方法。本研究结果表明,该番鸭三种细小病毒多联PCR检测方法具有特异、快速、准确、敏感的特点,能同时鉴别诊断这三种番鸭细小病毒,并可直接应用于临床典型病变组织的检测。

Three pairs of parvovirus generic primers were designed in this paper by using the VP and NS genes respectively from the GPV and MDPV in GenBank and from MDWNV in the Avian Disease Researchers of South China Agricultural University. Furthermore,the single PCR technology to optimize the reaction conditions of multi-PCR was established,such as the concentration of EX-Taq DNA polymerase enzyme,the primer concentration,the dNTPs concentration,the annealing temperature,and so on. The results showed that multi-PCR detection method of three parvovirus in the Muscovy ducks was specific,rapid,accurate and sensitive,and could synchronously differentiate three parvo virus of Muscovy ducks.Meanwhile,it could be used for the epidemic materials detection from the clinical typical pathological changes.