牛传染性鼻气管炎PCR检测方法的建立

Establishment of a PCR for Detection of Infectious Bovine Rhinotrachectis Virus

根据牛传染性鼻气管炎病毒gB基因序列设计一对引物,进行PCR检测,并对反应条件及反应体系进行优化。结果得到与预期大小(362bP)一致的特异性片段。最佳退火温度为58～64℃,Mg2+最佳浓度为1.6mM,dNTPS为0.36mM,引物为0.2pmol/uL,聚合酶0.8U/100uL。用这对引物扩增IBRV、HCV、PRRSV、PRV,只有IBRV扩增出特异性条带。该PCR方法的检测灵敏度为2.4ng/100uL,较其他方法敏感。

A pair of primers were designed for PCR according cheitis virus（IBRV）. The PCR assay was optimized and a special to the glycoproteinB（gB） gene of the Infectious bovine rhinotrapart of 362bp was obtained. The optimized annealing temperature was 58-64℃ and the optimized reaction system was 1.6mM Mg^2＋, 0.36mM dNTPs, 0.2 pmol/uL primers, 0.8U/100uL polymerase. Hog cholera virus（HCV）, Porcine reproduetive and respiratory syndrome virus（PRRSV）,Pseudorabies virus（PRV）and IBRV were amplified by the primers respeetively, only a special section of IBRV was obtained. The PCR assay ean deteet 2.4ng templet per 100uL and was sensitiver than other assays.