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猪瘟病毒4种检测方法的比较
张朝红1, 张彦明1, 张永国1
西北农林科技大学 动物科技学院
摘要:
应用病毒分离鉴定、荧光抗体法、反转录-聚合酶链式反应(RT-PCR)和夹心ELISA 4种方法,分别对23份猪瘟病料组织中的猪瘟病毒(CSFV)进行了检测,并比较4种检测方法的优缺点。结果显示,病毒分离鉴定法准确性较高,诊断比较容易,但存在试验步骤繁琐,所需时间长的缺点;荧光抗体检测法所需时间较短,但需要经验比较丰富人员来判定结果,且存在假阳性结果,特异性比较差;RT-PCR检测法具有快速、敏感等优点,特别是样品保存不理想时更有价值,利用套式PCR(nested-PCR)可以提高检测的特异性;夹心ELISA检测法具有快速、敏感等优点,但在样品保存不理想时会出现假阴性结果。
关键词:  猪瘟  病毒分离鉴定  荧光抗体  反转录-聚合酶链式反应  ELISA
DOI:
分类号:
基金项目:陕西省重大科技攻关项目(2006kz07-G2)
Comparisons of four methods for detection of classical swine fever virus
Abstract:
Four methods including the virus isolation and identification,fluorescent antibody,reverse transcriptase polymerase chain reaction(RT-PCR) and Sandwich ELISA were used to detect classical swine fever(CSFV) in 23 samples and were compared thereafter.The results showed that the virus isolation and identification could generate considerably reliable results but the procedure was complicated and costed long time;microtomy fluorescent antibody detection was fast,but the specificity was low and needed to exclude the false positive from the results.Sandwich ELISA detection combined the merits of the former two methods,but it generated the false negative results when the samples were in bad preservation.Compared with the others,RT-PCR was fast,high sensitive and especially useful for detecting the CSFV directly from the samples.The specificity could be raised by using the nested-PCR to detect the CSFV from the simples,especially when the simples were in bad preservation.
Key words:  classical swine fever  virus isolation and identification  fluorescent antibody  RT-PCR  ELISA