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猪流感RT-PCR诊断方法的建立
刘业兵
0
(中国兽医药品监察所)
摘要:
根据GenBank发表的多株猪流感病毒(SIV)序列,在保守区域设计1对引物,对RT-PCR反应条件进行优化,建立了可检测出多种亚型猪源流感病毒的RT-PCR方法。通过对反应产物测序分析,与预期结果相符,证明该方法具有很强的特异性;该方法可以检测出约1个TCID50的猪流感病毒,显示较高的敏感性。通过对12份临床样品进行检测,结果发现:该方法和其它两种方法分离病毒符合率相当。本研究建立的猪流感RT-PCR诊断方法具有高特异性和敏感性,适用于SIV的诊断应用和推广。
关键词:  猪流感病毒  RT-PCR  诊断
DOI:
投稿时间:2009-06-23修订日期:2009-10-23
基金项目:国家科技攻关计划十一五国家科技支撑计划 2007BAD86B04-4
The Establishing of RT-PCR Diagnostic Method for Swine influenza virus
liuyebing
(IVDC)
Abstract:
A pair of primers were designed from conserved regions according to the published swine influenza virus(siv) sequences in GenBank.The RT-PCR diagnostic method for swine influenza virus was established base on the optimization of RT-PCR reaction conditions.This method can specific detect a wide range of subtypes of swine influenza viruses. Sequence analysis of PCR products was in line with the expected results.This method has a sensitivity of detecting at least approximately 1 TCID50 swine influenza virus. Through 12 Clinical samples showed that the method was in line with the rate of using other two methods to separate virus. In this study, the establishment of the swine influenza diagnostic RT-PCR method has high specificity and sensitivity for the SIV diagnostic application and promotion..
Key words:  Swine influenza virus  RT-PCR  diagnosis

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