爲建立一種(zhǒng)适用于現場為好快速檢測的豬流行性腹瀉病毒（PEDV）LAMP技術，基于羟基萘酚藍（HNB）的可視化顯色特點，根據PEDV M基因編碼區序列，設計合成(chén樹算g)1套引物，通過(guò)反應物濃度和反應條件花雨優化，建立了可閉管檢測的PEDV RT-LAMP檢測方法。特異性和靈敏度試驗結那腦果顯示，建立的RT-LAMP檢測技術快速、靈敏、特異，可于1 h内檢出0.2 mL 0.1 TCID50/mL的病毒RNA，與實時(shí)熒光RT-PCR檢測方法靈敏度一緻，與豬瘟病毒、豬僞喝金狂犬病病毒、豬繁殖與呼吸綜合征病毒、豬圓環病毒機務2型以及豬鏈球菌2型核酸不發(fā)生交叉反應。利用該方法對(du明員ì)187份送檢的糞拭子及病死豬組織樣(yàng)品進(jìn)行應用檢測，檢出陽性樣短煙(yàng)品9份，與熒光定量RT-PCR方法檢測結果一緻。試驗結果表明，光微所建立的方法快速、特異，重複性滿足要求，海山适用于送檢樣(yàng)品的PEDV快速檢測。

Development of HNB-based Visual RT-LAM時輛P Method forDetection

of Porcine Epidemic討議 Diarrhea Virus

In order to est店司ablish a RT-LAMP me媽用thod forrapid detection of porcine epid慢員emic diarrhea viru著紅s（PEDV），based on color reaction of HNB a照報nd according to M genese友吃quence of PEDV，a set of primers were designed andsynt區農hesized. By optimi嗎線zing the reaction concentr看畫ation and conditions，the RT-LAMP method was deve懂子loped.Specificity and sensit姐鐵ivity results showed the establi街我shed method was fast，sensitive and specific. It co舞銀uld detectPEDV-RNA 黃友extracted from 0.2 mL v喝高irus suspension（0.1 TCID50/mL），which was consist姐兒ent with the real-time RT-PCR method.No地火 cross reactions were obs自習erved with the nucleic acids of c和美lassical swinefever virus（CSFV），pseudorabies virus（PRV），porcine reproductive and respirato短現ry syndrome vir師秒us（PRRSV），porcine circovirus type 2（PCV-2） and Streptococcus suis ty愛年pe 2. Using the RT-LAMPmethod，a total of 9 positive sample對一s were detectedfrom 187門身 samples of fecal swabs and dea做冷d pigs submitted，which was also consisten飛區t with the resultso頻通f real-time RT-PCR. As a 風劇conclusion，the established me知關thod was rapid，specific and repeatabl大用e，and it was suitable for  ra拿志pid detection of PEDV in submi街腦tted tissuesamples.

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