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Position: Home > Articles > Establishment of a PCR for Detection of Infectious Bovine Rhinotrachectis Virus China Animal Health Inspection 2006,23 (11) 26-28

牛传染性鼻气管炎PCR检测方法的建立

作  者:
徐淑菲;孔繁德;周斌华
单  位:
厦门出入境检验检疫局
关键词:
牛传染性鼻气管炎;引物;PCR
摘  要:
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。
译  名:
Establishment of a PCR for Detection of Infectious Bovine Rhinotrachectis Virus
作  者:
Xu Shu-fei, Kong Fan-de, Zhou Bin-hua (Xiamen Exit&Entry Enspection and Quarantine Service)
关键词:
IBRV PCR
摘  要:
A pair of primers were designed for PCR according to the glycoproteinB(gB) gene of the Infectious bovine rhinotracheitis virus(IBRV). The PCR assay was optimized and a special part of 362bp was obtained. The optimized annealing temperature was 58~64℃ and the optimized reaction system was 1.6mM Mg2+, 0.36mM dNTPs, 0.2 pmol/uL primers, 0.8U/100uL polymerase. Hog cholera virus(HCV), Porcine reproductive and respiratory syndrome virus(PRRSV),Pseudorabies virus(PRV)and IBRV were amplified by the primers respectively, only a special section of IBRV was obtained. The PCR assay can detect 2.4ng templet per 100uL and was sensitiver than other assays.

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