【摘要】：為了建立一種快速、特異、靈敏的檢測(cè)I群禽腺病毒的液相基因芯片方法。根據(jù)Hexon基因序列設(shè)計(jì)特異引物,上游引物5'端加TAG序列,下游引物5'端加生物素,進(jìn)行PCR擴(kuò)增,將擴(kuò)增產(chǎn)物與鏈霉素親和蛋白、磁珠37℃孵育30 min,通過(guò)Luminex200儀器分析。用所建立的液相基因芯片方法進(jìn)行特異性、靈敏度、重復(fù)性及病料樣品的檢測(cè)。結(jié)果該方法的特異性強(qiáng);檢測(cè)的敏感性可達(dá)1×102copies/μL;批內(nèi)批間的變異系數(shù)都在5%以下;檢測(cè)結(jié)果與SYBR Green I熒光PCR方法 100%相符。表明建立的液相基因芯片方法特異性好、靈敏高、重復(fù)性好,可用于I群禽腺病毒的檢測(cè)。
[Abstract]:In order to establish a rapid, specific and sensitive liquid-phase microarray method for detection of group I avian adenoviruses. According to the Hexon gene sequence, specific primers were designed. The upstream primer 5 'end added TAG sequence and downstream primer 5' end added biotin. The amplified product was amplified by PCR. The amplified product was incubated with streptomycin affinity protein at 37 鈩，

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