鸡传染性喉气管炎病毒PCR诊断方法的建立与应用

根据GenBank登录的传染性喉气管炎病毒(ILTV)的TK基因序列设计并合成1对特异性引物,以ILTV疫苗株DNA为模板,建立了检测ILTV TK基因的PCR方法。应用该方法能从临床分离毒株和疫苗株中扩增到长为427 bp的目的片段;但不能从新城疫病毒(NDV)、传染性法氏囊病毒(IBDV)、禽呼肠孤病毒(ARV)、减蛋综合征病毒(EDSV)、H9亚型禽流感病毒(H9-AIV)、传染性支气管炎病毒(IBV)、大肠杆菌以及金黄色葡萄球菌等病原中扩增出阳性条带;敏感性试验表明其DNA最小检出量为4.9 ng;应用该方法和病毒分离法对2份临床病例和人工感染鸡的检测,两者符合率为100%。上述结果表明该PCR方法具有良好的特异性和敏感性,可用于传染性喉气管炎病毒鉴定和临床诊断。

Establishment and application of polymerase chain reaction for infectious laryngotracheitis virus

According to the TK gene sequence of infectious laryngotracheitis virus (ILTV) in GenBank, a pair of primers were designed and synthesized. Using the genomie DNA of ILTV vaccine strain as a template, a PCR method was established for the detection of ILTV TK gene. The target fragment of 427 bp could be amplified from the ILTV strains and vaccine strains, but not from the Newcastle disease virus (NDV), infectious bursal disease virus (IBDV), avian reovirus (ARV), egg drop syndrome virus (EDSV), H9 subtype of avian influenza virus (H9-AIV), infectious bronchitis virus IBV, Escherichia coli and Staphylococcus aureus. The sensitivity test showed that the minimum detectable amount of DNA was 4. 9 ng. The PCR method and virus isolation were used on 2 clinical cases and the artificially infected chickens, and the coincidence rate between the two methods was 100%. The results indicated that the PCR method has good specificity and sensitivity, and can be used for the detection of ILTV.