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禽流感病毒實時熒光定量RT-PCR檢測方法的建立

點擊數:15683  時間:2019-05-10 13:39:09  來源: 中國動物檢疫      作者: 王楷宬,王素春,朱琳等

為滿足禽流感病毒高通量快速檢測的需要,建立了一種能夠檢測各亞型禽流感病毒的實時熒光定量RT-PCR檢測方法,并對該方法的特異性和靈敏度進行評估,使用該方法與農業行業标準NY/T772—2013中的禽流感病毒RT-PCR方法同時進行臨床樣品檢測。結果顯示,該方法檢測耗時短、特異性好,檢測下限達10-4 ng/μL,與傳統的RT-PCR方法陽性符合率為100%。結果表明,本方法能實現對禽流感病毒的安全、特異、快速、靈敏、簡單、高通量檢測,從而彌補了現有傳統檢測技術的不足。



Establishment of Real-time RT-PCR for Detection of AvianInfluenza Virus

In order to achieve the high-throughput and rapid detection ofavian influenza virus(AIV),a real-time fluorescent quantitative RT-PCR method was establishedfor detecting all subtypes of AIV. Then its specificity and sensitivity wereevaluated,and some clinical samples were testedsimultaneously by the established method and the conventional RT-PCR stipulatedin the agricultural industry standard NY/T772—2013. Results showed that thedetection limit of the established method could reach 10-4 ng/μL with shortdetection time and better specificity,and the positivecoincidence rate was 100% as compared with traditional RT-PCR method. Inconclusion,the established real-time RT-PCR method wassafe,specific,rapid,sensitive and simple,and could achieve thehigh-throughput detection for AIV,which covered thedeficiency of current traditional detection technologies.




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