肉牛附红细胞体PCR检测方法的建立与初步应用

Establishmen and Application of PCR Method for Detection of Cattle Eperythrozoonosis

为建立一个能快速准确的检测出肉牛附红细胞体的分子生物学诊断方法,及时监控好人畜共患病。试验根据Genbank中发表的肉牛附红细胞体16SrRNA序列设计一对特异性引物,通过PCR方法,对肉牛附红细胞体基因组DNA进行扩增。结果表明:扩增出了一段412 bp的DNA片段,与Genbank中发表的肉牛附红细胞体序列同源性为98%。将建立的PCR方法对大肠杆菌、弓形体、胸膜肺炎放线杆菌的DNA进行检测为阴性,检测肉牛附红细胞体DNA的最低浓度为1.23 ng/μL。该方法快速、准确、特异性好、灵敏度高,为肉牛附红细胞体病的临床诊断提供了新的方法。

In order to establish a diagnostic method of molecular biology that can rapidly and accurately detect cattle Eperythrozoon to survey and prevent and control zoonoses, a pair of specific primers were de- signed according to the published sequence data 16s ribosomal RNA gene of beef cattle Eperythrozoon from Genbank to amplify genomic DNA of cattle Eperythrozoon. The result showed that the targeted gene fragment was 412 bp in length and 98% identical to the published data. The genomic DNA of Escherichia coli, Toxo- plasm and Actinobacillus pleuropneumoniae ity of the assay was 1.23 ng/μL of DNA. was amplified by the method, the result was negative. The sensitiv- It suggested that the method is a rapid, accurate, specific and sen- sitive diagnostic method for cattle Eperythrozoon.