為建立一種適用于現(xiàn)場快速檢測的豬流行性腹瀉病毒（PEDV）LAMP技術(shù),，基于羥基萘酚藍(lán)（HNB）的可視化顯色特點(diǎn),，根據(jù)PEDV M基因編碼區(qū)序列,，設(shè)計(jì)合成1套引物,，通過反應(yīng)物濃度和反應(yīng)條件優(yōu)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(yàn)結(jié)果顯示,，建立的RT-LAMP檢測技術(shù)快速,、靈敏、特異,，可于1 h內(nèi)檢出0.2 mL 0.1 TCID50/mL的病毒RNA,，與實(shí)時(shí)熒光RT-PCR檢測方法靈敏度一致,，與豬瘟病毒、豬偽狂犬病病毒,、豬繁殖與呼吸綜合征病毒,、豬圓環(huán)病毒2型以及豬鏈球菌2型核酸不發(fā)生交叉反應(yīng)。利用該方法對187份送檢的糞拭子及病死豬組織樣品進(jìn)行應(yīng)用檢測,，檢出陽性樣品9份,，與熒光定量RT-PCR方法檢測結(jié)果一致。試驗(yàn)結(jié)果表明,，所建立的方法快速,、特異，重復(fù)性滿足要求,，適用于送檢樣品的PEDV快速檢測,。

Development of HNB-based Visual RT-LAMP Method forDetection

of Porcine Epidemic Diarrhea Virus

In order to establish a RT-LAMP method forrapid detection of porcine epidemic diarrhea virus（PEDV），based on color reaction of HNB and according to M genesequence of PEDV,，a set of primers were designed andsynthesized. By optimizing the reaction concentration and conditions,，the RT-LAMP method was developed.Specificity and sensitivity results showed the established method was fast，sensitive and specific. It could detectPEDV-RNA extracted from 0.2 mL virus suspension（0.1 TCID50/mL）,，which was consistent with the real-time RT-PCR method.No cross reactions were observed with the nucleic acids of classical swinefever virus（CSFV）,，pseudorabies virus（PRV），porcine reproductive and respiratory syndrome virus（PRRSV）,，porcine circovirus type 2（PCV-2） and Streptococcus suis type 2. Using the RT-LAMPmethod,，a total of 9 positive samples were detectedfrom 187 samples of fecal swabs and dead pigs submitted，which was also consistent with the resultsof real-time RT-PCR. As a conclusion,，the established method was rapid,，specific and repeatable，and it was suitable for  rapid detection of PEDV in submitted tissuesamples.

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