猪伪狂犬病病毒套式PCR检测方法的建立与应用  

Establishment and Application of nPCR for Detecting Porcine Pseudorabies Virus

作　　者：吴旭锦[1] 朱小甫[1] 

机构地区：[1]咸阳职业技术学院畜牧兽医研究所动物疫病分子生物学诊断实验室,陕西咸阳712000

出　　处：《动物医学进展》2016年第6期18-21,共4页Progress In Veterinary Medicine

基　　金：咸阳市科学技术研究计划项目(2015k03-21)

摘　　要：对伪狂犬病病毒(PRV)国标PCR方法进行优化,建立一种高灵敏度的套式PCR方法。根据伪狂犬病病毒gD基因序列,设计并合成了2对引物,通过反应体系和条件的优化建立套式PCR方法。通过灵敏性试验、特异性试验、病料检测对比试验等验证建立方法的适用性。结果表明,建立的方法检测伪狂犬病病毒DNA的极限为2.6×10-7 ng/mL,灵敏度比国家标准方法提高了1 000倍,从疑似PRV感染组织病料中能大幅度提高阳性检出率。本研究成功建立了一种灵敏度高、特异性好的快速检测猪伪狂犬病病毒的套式PCR检测方法。To establish a highly sensitive nested PCR methods for detecting porcine pseudorabies virus,according to published gene sequences of PRV gD,two pairs of primers were designed and synthesized,the nested PCR method was established by optimizing the reaction system and conditions.The method applicability was verified by sensitivity test,specificity test,sample comparison tests.The results showed that the detection limit of the method established is 2.6 × 10-7 ng/mL,the sensitivity is 1 000 times higher than that of national standard method,and the objective band was only amplified from the pseudorabies virus reference strain,and could greatly improve the positive detection rate from suspected samples.The nested PCR detection mehod with high sensitivity and good specificity for rapid PRV detection was successfully established.

关 键 词：伪狂犬病病毒 套式PCR 诊断 应用 

分 类 号：S852.65[农业科学—基础兽医学]