基于钙黄绿素显色的可视化LAMP检测猪流行性腹泻病毒的研究

doi:10.16431/j.cnki.1671-7236.2015.02.014

Abstract

Abstract: In order to establish a loop-mediated isothermal amplification assay (LAMP),a set of LAMP primers targeting to NP gene of porcine epidemic diarrhea virus (PEDV) were designed and synthesized.Calcein and manganese chloride were added as the indicator instead of SYBR Green Ⅰ after reaction.The LAMP could amplify PEDV specifically at 65 ℃ in 1 h with detection limit of 3.73 pg/μL and the result could be judged by naked eye.The LAMP didn't react with CSFV,TGEV,PoRV and E.coli.93 clinical samples of PED suspected case collected from Veterinary Institute of Longyan University were submit to detect PEDV by LAMP,the result showed that the positive rate of the samples was 43.01%,which was higher than that (38.7%) of conventional PCR.The established visual LAMP could be used for diagnosing PED.

Key words: loop-mediated isothermal amplification assay (LAMP); porcine epidemic diarrhea virus (PEDV); calcein; diarrhea; detection

CLC Number:

S852.65

TANG Xiao-zhen, CHEN Lu-wei, LU Rong-bin, ZHENG Xin-tian, HUANG Cui-qin. Detection of Porcine Epidemic Diarrhea Virus by Calcein-based Visual Loop-mediated Isothermal Amplification (LAMP) Assay[J]. , 2015, 42(2): 331-336.

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Detection of Porcine Epidemic Diarrhea Virus by Calcein-based Visual Loop-mediated Isothermal Amplification (LAMP) Assay

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