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      1. 國(guó)家獸藥產(chǎn)業(yè)技術(shù)創(chuàng)新聯(lián)盟
        National veterinary drug industry technology innovation alliance
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        H9亞型禽流感病毒實(shí)時(shí)熒光RT-PCR檢測(cè)方法的建立與應(yīng)用

        時(shí)間:2019-12-16   訪問量:1045


        為建立一種快速準(zhǔn)確檢測(cè)H9亞型禽流感病毒基因的檢測(cè)方法,根據(jù)GenBank中H9亞型禽流感病毒血凝素編碼基因進(jìn)行熒光檢測(cè)引物和探針設(shè)計(jì),建立了一種針對(duì)H9亞型禽流感病毒的熒光RT-PCR檢測(cè)方法。利用該方法進(jìn)行靈敏度和特異性檢測(cè),并用本方法和國(guó)家標(biāo)準(zhǔn)中的熒光RT-PCR檢測(cè)方法,同時(shí)對(duì)臨床樣本進(jìn)行檢測(cè)。結(jié)果顯示,僅H9亞型禽流感病毒出現(xiàn)了正常熒光檢測(cè)曲線,而其他病毒及陰性對(duì)照均未出現(xiàn);檢測(cè)靈敏度為RNA終濃度10-4 ng/μL(1.30×104 copies/μL);臨床樣本檢測(cè)結(jié)果與國(guó)標(biāo)方法一致,符合率為100%。結(jié)果表明,該方法特異性強(qiáng)、靈敏度高,可用于H9亞型禽流感病毒檢測(cè)。

        Establishment and Application of a Real-time RT-PCR Assay for Detection

        of H9 Subtype Avian Influenza Virus

        In order to establish a method for detecting H9 subtype avian influenza virus(AIV-H9)rapidly and accurately,the fluorescence detection primers and probes were designed and synthesized according to hemagglutinin encoding gene of AIV-H9 registered in GenBank,and a fluorescence PCR assay was thereby established,and its specificity and sensitivity were tested. Then by the established assay and the RT-PCR stipulated in national standard,some clinical samples were simultaneously tested. The results showed that the normal fluorescence detection curve only appeared in AIV-H9 rather than other non-specific viruses and negative sample;its detection limit was 10-4 ng/μL(1.30×104 copies/μL)of RNA;the detection result of clinical samples by established assay was consistent with the national standard,and the coincidence rate was 100%. In conclusion,the assay could be used to detect AIV-H9 due to its strong specificity and high sensitivity.

        全文下載鏈接:https://kns.cnki.net/KCMS/detail/37.1246.S.20191206.0907.024.html











        國(guó)家獸藥產(chǎn)業(yè)技術(shù)創(chuàng)新聯(lián)盟
        National veterinary drug industry technology innovation alliance

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