Establishment and Application of Loop-mediated Isothermal Amplification for Rapid Detection of Porcine Epidemic Diarrhea Virus |
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| Authors: | LUO Ya-kun LIANG Lin WANG Jing LIU Cun LIU Qi LIU Chang LIN Wen-cheng CUI Shang-jin |
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| Institution: | 1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Beijing Observation Station for Veterinary Drug and Veterinary Biotechnology, Ministry of Agriculture, Beijing 100193, China;
3. College of Animal Science, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | This study was aimed to establish a loop-mediated isothermal amplification (LAMP) assay for detection of porcine epidemic diarrhea virus (PEDV), and provide a simple, sensitive, accurate and reliable tool for diagnosis of PEDV.The conservative PEDV N gene (GenBank accession number: KT799997) of PEDV was selected as a target to design six specific primers.The reaction system and temperature of LAMP were optimized, and the LAMP method for specific amplification of PEDV was established. Results showed that the PEDV LAMP detection method was established successfully, and it could detect PEDV specifically at 60℃ for 60 min,and the detection limit was 91 copies/μL, which was one hundred-fold higher than conventional RT-PCR method.75 clinical samples were detected by LAMP and PCR, respectively, the coincidence of LAMP and PCR was about 97.3%. All the data suggested that the LAMP assay had strong specificity, high sensitivity, simple operation, low equipment requirement, and was suitable for rapid detection of PEDV clinical samples. |
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| Keywords: | porcine epidemic diarrhea virus (PEDV) loop-mediated isothermal amplification (LAMP) detection |
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