狂犬病毒、僞狂犬病毒、犬瘟熱病毒小黃、犬細小病毒和犬冠狀病毒是引起(qǐ)術店犬科動物重要傳染病的5種(zhǒng)病毒，嚴重威脅著舊農(zhe)犬科動物的健康，是出入境犬科動物重點篩查的緻病照著因子。爲提升口岸進(jìn)出境犬科動物檢疫工作效時音率，建立了上述5種(zhǒng)病毒微流控芯片檢測方法。在建立5種信司(zhǒng)病毒環等溫擴增檢測技我見術（loop mediated is聽站othermal amplification，L金司AMP）的基礎上，將(jiāng)這(zhè)些LAMP檢測體系固化數火在同一塊塑料芯片上，制備了同步高通量快速檢測微在紙流控芯片，并將(jiāng)其應用于臨床近短檢測。結果顯示：微流控芯片檢測方法具有良好(hǎo頻藍)的特異性，含有目的基因的陽性樣(yàng)本你務僅在芯片上相應病毒反應槽出現顯著擴增，而其他病毒反應槽未出現擴增，彼此無交街媽叉反應；微流控芯片的敏感性與LAMP方法一緻；通過(gu問道ò)檢測不同時(shí)期收集近文的感染犬瘟熱病毒、犬細小病毒和犬冠狀病毒臨床樣(yàng)本錢水，證實建立的微流控芯片檢測方法具有良好(hǎo)的穩定性和可靠大村性。與現有核酸檢測方法相比，微流控芯片可以在受試核酸上樣(y兒火àng)40 min内完成(chéng)上述5種(zhǒ和他ng)犬病毒的快速篩查，從而滿足進(jìn)出境犬科動物快速檢疫的需笑吧求。
Establishment and Ap湖鐵plication of the Microf和湖luidic Chip Assay for Five Kinds o工錢f Major Canine Viruse開慢s
Rabies virus，pseudorabie區國s virus，canine distemper virus，c器化anine parvovirus and canine cor花亮onavirus are five kinds of viruse村分s that cause important infecti大公ous diseases of canine，which seri船音ously threaten the health 林嗎of canine，and ar行紅e the key pathogenic f們但actors to be screened and exami路物ned by the exit-entry ports. In跳員 order to improve the efficiency of por體音t entry-exit quarantine for canine，a mi看數crofluidic chip assay for t術鐵he above five viruses was esta白師blished. Five loop-mediated音飛 isothermal amplification（LAMP）assays w著木ere specifically e雪報stablished and then so書問lidified on the生村 same plastic chip to prepare a mic暗學rofluidic chip with hig遠劇h-throughput and rapid detection科空 capacity for clinical detectio和他n. The results showed that the est白線ablished assay had a 人些good specificity where 服商positive samples 街那with targeted genes were obviousl秒音y amplified only 話場in the reaction tank in correspondin唱問g viruses instead of othe間是r viruses，and no c算道ross reaction with each other was分民 found；the sensitivi是司ty of microfluidic 術謝chip was consistent with that of L紙吃AMP；the establis家理hed assay was with good stab湖樹ility and reliability as verifi藍離ed through detecting clinic藍答al samples infected with canine公哥 distemper virus身他 and canine parvovirus and collected音又 in different periods.看暗 Compared to existing nucleic acid de地算tection methods，the媽行 microfluidic chip could be used to co是匠mplete rapid sc費外reening of the above five viruse得場s within 40 min拍化utes after the loading物鄉 of nucleic acids，which could綠理 satisfy the needs of rapid 醫草quarantine for entry-exit canine ani但從mals.
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