Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a Campylobacter jejuni Clone
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概要
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Loop-mediated isothermal amplification (LAMP) assay is a simple, rapid and specific detection method and has been used for detection and identification of different Campylobacter species. In this study, we develop a LAMP assay specific for detection of a particular clone (clone SA) of Campylobacter jejuni, associated with the vast majority of recent sheep abortions in the U.S. Using a set of specific primers for C. jejuni IA3902 (a clone SA isolate) and genomic DNA or boiled cell extract as template, the target DNA was amplified at 63°C for 50 min in a water bath. A positive reaction was identified visually as white precipitate or fluorescence under UV, and confirmed by gel electrophoresis. Detection limit of the assay was comparable to that of conventional PCR. The LAMP was shown to be specific for detection of clone SA when tested on a number of C. jejuni strains of different genetic backgrounds. Applicability of the LAMP assay for specific detection of clone SA was demonstrated in animal tissues experimentally infected with IA3902 or genetically diverse C. jejuni strains. Since clone SA is the predominant cause of sheep abortions in the U.S. and is a zoonotic pathogen, the LAMP assay may be a valuable detection tool in future epidemiological studies.
著者
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Luo Yan
Department Of Child Health Care The Children's Hospital Of Zhejiang University School Of Medici
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LUO Yan
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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SAHIN Orhan
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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DAI Lei
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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SIPPY Rachel
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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WU Zuowei
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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ZHANG Qijing
Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.
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