【摘要】 为研究一种可同时检测鸡传染性贫血病毒(CIAV)和鸡细小病毒(ChPV)的双重PCR方法,本试验根据GenBank中CIAV的VP基因和ChPV的NS1基因的保守序列,设计筛选了扩增片段大小分别为219 bp和384 bp的特异性引物,通过对反应条件的优化,特异性和敏感性试验,建立和评价双重PCR检测方法。结果显示,该方法最佳引物比例为CIAV上下游引物各0.4μL(20μmol/L),ChPV上下游引物各0.6μL(20μmol/L);最佳退火温度为56.1℃;灵敏度可达到66 fg(CIAV)和78 fg(ChPV);对常见鸡病病原体进行检测,结果全为阴性。结果表明,本试验所建立的双重PCR方法具有特异、敏感、快速、稳定等优点和优势,可用于同时快速鉴别诊断CIAV和ChPV的混合感染。更多还原

【Abstract】 The aim of this research was to develop a duplex PCR assay for detecting both chicken infectious anemia virus(CIAV) and chicken parvovirus(ChPV). According to the conserved gene sequences of CIAV VP gene and ChPV NS1 gene in GenBank, two sets of specific primers were designed. The size of the amplified fragment were 219 bp and 384 bp. The duplex PCR assay was established and evaluated by optimizing the reaction conditions, specificity and sensitivity test. The result showed that the best primers were 0.4 μL(20 μmol/L) for CIAV and 0.6 μL(20 μmol/L) for ChPV. The optimum annealing temperature was 56.1 ℃.The sensitivity of the assay was 66 fg for CIAV and 78 fg for ChPV. The pathogens of common chicken diseases were detected, and the reasults were all negative. This duplex PCR assay is a quick, sensitive, and specific test for detection of CIAV and ChPV,and will be useful for the accurate diagnosis of these viruses in chickens.更多还原