【摘要】 〔目的〕建立牛传染性鼻气管炎的PCR检测方法。〔方法〕根据牛传染性鼻气管炎病毒的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0.2 pmol/μL,聚合酶0.8U/100μL。用这对引物扩增IBRV、HCV、PRRSV、PRV,只有IBRV扩增出特异性条带。〔结论〕该PCR方法的检测灵敏度为2.4ng/100μL,较其他方法敏感。更多还原

【Abstract】 A pair of primers were designed for PCR detection 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 p mol/μL primers,0.8U/100μL 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 100μL and is more sensitive than other assays.更多还原