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一种用于马立克氏病病原检测的PCR技术研究

     

摘要

Specificity and sensitivity comparative studies were carried out in a pair of self-designed prim-ers and four pairs of published PCR detection primers .The PCR products were sequenced and analyzed .All the primers were found to be highly specific ,but their sensitivities were different .A target bang could be amplified when the DNA concentration reached 3.81×10-3 μg/mL,so the sensitivity of Meq 5 primer was better than that of other four pairs of primers .Sequencing analysis results showed that the SD 2012-1 have high homology with other reference strains .Detection result of tissue samples showed that Marek ’ s disease virus can be detected in feather pulp,heart,liver,spleen,kidney,glandular and duodenum .This study provides a basis for epidemiologi-cal investigation and detection of Marek ’ s disease .%根据马立克氏病Meq基因序列特点设计的一对检测引物,与其他4对已发表的检测引物进行特异性和敏感性的比较研究,对PCR产物进行测序分析。结果显示,5对PCR检测引物均具有较好的特异性,但敏感性不完全相同,自行设计的Meq5引物在DNA浓度为3.81×10-3μg/mL时仍能够扩增出目的条带,明显优于其他4对引物。序列测定结果显示,新的分离株SD2012-1与国内外其他参考毒株之间同源性较高。组织样品检测结果显示,羽髓、心脏、肝脏、脾、肾脏、腺胃及十二指肠病料中均能检测到马立克氏病病毒,为今后马立克病的流行病学调查和检测提供了依据。

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