鸡肿瘤病快速鉴别诊断技术的建立

THE DEVELOPMENT OF RAPID DIFFERENTIAL DIAGNOSIS TECHNIQUE FOR AVIAN TUMORS

根据病毒基因组特征,以聚合酶链反应(PCR)为基础,建立一种能快速鉴别马立克氏病(MD)、禽白血病(AL)和网状内皮增生症(RE)3种常见鸡肿瘤病的诊断技术。研究确定以MDV的132 bpr及ALV和REV的LTRs作为PCR扩增的特异性基因片段,改进样品DNA的提取程序,优化PCR及三重PCR的反应条件及参数,并对该诊断技术的特异性、敏感性进行研究。结果表明:建立的诊断技术具有特异、敏感、快速、经济的优点,为开发鸡肿瘤病快速鉴别诊断试剂盒奠定了基础。

Marek′s disease (MD), reticuloendotheliosis (RE) and avian leukosis (AL) are the most common tumors in chickens. A rapid differential diagnosis for avian tumors, which based on PCR technique was developed to amplify the specific gene fragments: 132bpr of MDV, LTRs of ALV and REV viral genomes. Improved protocol for sample DNA extraction, optimized condition and parameters for PCRs were approached. In addition, the specificity and the sensitivity of the PCRs were also evaluated. The whole procedure, from sampling to the final conclusion, was finished within one day′s workhour (8 h) in our protocol. The results demonstrated that the PCR protocol is a specific, sensitive and rapid tool for the differential diagnosis of avian tumors, which provides a possibility for the development of rapid differential detection kit for avian tumors.