爲建立可以區分H7N9流感病毒強毒株和弱毒株的方法，通過(guò光個)比對(duì)GenBank和GISA也鄉ID中H7N9流感病毒的HA基因序列雜師以及本實驗室保存的病毒序列，設計1對(duì)特異性引物和2條探針鐘藍，建立了區分H7N9流感病毒強弱毒株的實時(s船謝hí)熒光RT-PCR方法，同時(shí)對(體為duì)該方法的反應體系和反應參數進(jìn)行優化相紅，并開(kāi)展特異性和敏感性試驗以及臨床應用試驗。特異性試飛唱驗顯示，該方法具有良好(hǎo)的特異性，對(duì)其他常章有見亞型的禽流感病毒和其他禽病病毒檢測均爲農東陰性。敏感性試驗顯示，該方法對(duì)H7N9流感制高強毒和弱毒HA基因的檢測下限分别爲0.004 fg 師來和0.1 fg RNA模闆，高于兩(l是錢iǎng)種(zhǒng)商品化試劑舊身盒的靈敏度。利用該方法對(duì)40株經(jīng)實驗票熱室分離鑒定的H7N9流感病毒進(jìn)行對(duì)比驗證，發(fā)現檢討劇出率和符合率均爲100%。結果表明，該方法具有特異、敏感、準确的優點老行，可用于H7N9流感病毒的快速檢測，同時秒呢(shí)還(hái)可區分強知場弱毒株，對(duì)H7N9流感病毒，尤其是高緻病性流感病毒的早期診斷可站和有效防控具有重要意義。
Development of a Real-time RT-PCR Met開明hod
for Identification of Virulent and Avir服道ulent Strains of H7N9Influenz了雪a Virus
In order to develop a method for id司麗entifying virulent anda站生virulent strains of H7N9 inf件關luenza virus，a pair ofspecific primers店計 and probes were desi門司gned by comparing the HA 司姐gene sequences ofH7N9 i黑事nfluenza virus 和還in Genbank and G校請ISAID and the sequ日長ence of which wasiso請習lated and sequenced by ou小錯r laboratory，a real-timeRT-PCR m報離ethod was developed，me場他antime，the reaction system and女國 parameters of the method were opti愛你mized，and the specificity a南我nd sensitivity as well as its clinical兒下application were tested. T跳兒he specificity test showed th做信at，the developmen明市t method didn't react務器 with other common subtype都睡s ofavian influenza virus花術 and other avian鐘工 virus，showing a森聽good specificity. Sensitivity test show問小ed that，by theRT-PCR method，the lower 間從detection limits of HA genes ofviru土藍lent and attenu船大ated H7N9 strains有報 were 0.004 fg and 0.1 f相票g respectively，which were all lower th鄉現an the detection limits市要 of the two kinds 雜關ofcommercial kits. By comparing and v東高erifying 40 strains of H7區道N9 influenza virusisolated a老好nd identified by the laboratory，it was 也對foundthat both the detect來不ion rate and coincidence rate were 農村100%. Therefore, it wasco司可ncluded that the method 家兒could be used for rapid雨些 detection of H7見到N9 virus andiden飛老tifying the virule大熱nt and avirulent strains due to its理明 advantages of 綠民goodspecificity，sens去這itivity and high accuracy，whic秒空h was of great signif南刀icance for early dia很關gnosis and effecti路新veprevention and control of H7N9 virus，雨水especially thehighly pathogenic 內從virus.
全文下載鏈接：
http://kn我通s.cnki.net/KCMS/detail/detail.as綠中px?dbcode=CJFQ&dbname=CJFDPREP&filena熱森me=ZGDW201904015&v=MDA0NjZPZlkrUnF又醫GeS9rVWJySlB5clBlYkc0購很SDlqTXE0OUVZWVI4ZVgxTHV4WVM3R冷費GgxVDNxVHJXTTFGckNV暗房Ukw=
