Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive and Specific Detection of a Campylobacter jejuni Clone

Journal of Veterinary Medical Science

Yan LUO¹  Lei DAI¹  Rachel SIPPY¹  Zuowei WU¹  Qijing ZHANG¹  Orhan SAHIN¹ 

[1] Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, Iowa 50011, U.S.A.

关键词: Campylobacter jejuni clone SA;    detection;    loop-mediated isothermal amplification;    PCR;   

DOI  :  10.1292/jvms.11-0462

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(19)Cited-By(1)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.

【 授权许可】

Unknown