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Development of loop-mediated isothermal amplification and PCR assays for rapid and simple detection of Campylobacter
Wataru Yamazaki1, Masumi Taguchi, Naoaki Misawa
1Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, Miyazaki, Japan. yamazakiw@cc.miyazaki-u.ac.jp
Abstract:
Campylobacter fetus is divided into CFV and CFF. Because CFV causes bovine genital campylobacteriosis, differentiation of the two subspecies is essential to the implementation of efficient CFV control and eradication programs. We have developed LAMP and duplex PCR assays for rapid and simple detection of CFV. The LAMP assay correctly detected 7 CFV strains and did not detect 53 CFF, 35 non-fetus Campylobacter and 25 non-Campylobacter strains. The PCR assay successfully differentiated the two subspecies. The LAMP and PCR assays were faster than conventional biochemical assays, requiring for detection less than 50 min and less than 4 hr, respectively, from the beginning of DNA extraction from a single colony on blood agar to final determination. Our LAMP and PCR assays are rapid and practical tools for detection of CFV.
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