H5亞型禽流感病毒HA序列差異越來越大。爲更準确地檢測H5白計亞型禽流感病毒，對(duì)GenBank中發(fā)表的H5亞型禽流感病毒H樹用A序列以及本實驗室保存病毒序列進(j議北ìn)行比對(duì)，同時(shí)設計1對(du船雨ì)引物和1條探針，建立了H5亞型禽流感病毒實呢小時(shí)RT-PCR方法，并對(duì)該方法的反應體系和反應參數進(jì的快n)行了優化。以本實驗室分離測序确定的30份H5亞型禽流感病毒RNA爲模闆，將費事(jiāng)該方法與兩(liǎng)種(zhǒng)商品化H爸金5亞型禽流感病毒檢測試劑盒進(jìn)行比對村低(duì)，發(fā)現該方法與兩(liǎ刀請ng)種(zhǒng)商品化試劑盒的檢出做會率分别爲100%、98%、98%。敏感性試車光驗顯示，該方法可以檢測出0.1 fg的RNA模闆，靈敏度比兩(liǎng)種微算(zhǒng)商品化試劑盒均提高了1音說0倍。結果表明，該方法具有更高的特異性和敏感性，可用于H5亞型信腦高緻病性禽流感的早期診斷。

Establishment and Application of計身 Real-time RT-PCR for Detect嗎聽ion

of H5 Subtype Avian I文東nfluenza Virus

In recent years機去，thedifference in HA動物 sequence of H5 subtype avia務窗n influenza virus（AIV）was著嗎 increasing. In order to 制科detect H5subtype AIV m動計ore accurately，暗飛by comparing the HA開務sequence of H5 subt刀術ype AIV published in GenBank with the車有 one conserved in thelabo自月ratory and designing a pair of pri道裡mers and one TaqMan probe，a real-time地土 RT-PCR method wa影車s developed，andits reaction 玩劇system and paramet唱門ers were optimized. Taking 30 RNA sam吧紙ples of H5subtype AIV strain 習店isolated and sequen電照ced by the laborato紙場ry as the template，the established me街體thod was compared with two commerci兵制al H5 subtypeAIV detection匠也 kits，and it was如兵 found that the detection車低rates were 100%，98% and 98% respecti空事vely. Sensitivitytest showed t紙廠hat RNA template of 0.1 fg could be筆廠 detected by the懂哥 establishedmethod w藍資hose sensitivity was 10 tim化短es higher than that of 小年the two commercialkits. In conc河討lusion，the RT-PCR method was 火鐵more specificand sensiti的了ve，and it could 行通be used for early diagnosisof H5 subtyp她文e highly pathogenic avian influe裡愛nza.

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