Development of a loop-mediated isothermal amplification assay for sensitive and rapid detection of Campylobacter fetus

We developed a sensitive and rapid loop-mediated isothermal amplification (LAMP) assay for detection of Campylobacter fetus. This assay provides simpler and more rapid detection of C. fetus than conventional biochemical and PCR assays. The assay correctly detected 60 C. fetus strains but not 55 non-...

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Veröffentlicht in:	Veterinary microbiology 2009-05, Vol.136 (3), p.393-396
Hauptverfasser:	Yamazaki, Wataru, Taguchi, Masumi, Ishibashi, Masanori, Nukina, Masafumi, Misawa, Naoaki, Inoue, Kiyoshi
Format:	Artikel
Sprache:	eng
Schlagworte:	accuracy animal tissues Animals Bacteriology bioassays Biological and medical sciences Campylobacter fetus Campylobacter fetus - genetics Campylobacter fetus - isolation & purification Campylobacter Infections - diagnosis Campylobacter Infections - microbiology campylobacteriosis Cattle cattle diseases Cattle Diseases - diagnosis Cattle Diseases - microbiology disease detection disease diagnosis DNA - chemistry DNA - genetics Female food pathogens Fundamental and applied biological sciences. Psychology human diseases Liver - microbiology loop-mediated isothermal amplification Loop-mediated isothermal amplification (LAMP) assay Microbiology Miscellaneous molecular epidemiology new methods Nucleic Acid Amplification Techniques - methods Nucleic Acid Amplification Techniques - veterinary nucleic acids pathogen identification polymerase chain reaction rapid methods Sensitive and rapid detection Sensitivity and Specificity Vagina - microbiology validity
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Zusammenfassung:	We developed a sensitive and rapid loop-mediated isothermal amplification (LAMP) assay for detection of Campylobacter fetus. This assay provides simpler and more rapid detection of C. fetus than conventional biochemical and PCR assays. The assay correctly detected 60 C. fetus strains but not 55 non- fetus Campylobacter and 30 non- Campylobacter strains. The sensitivity of the LAMP assay in pure cultures and in a spiked bovine liver specimen was 10-fold more sensitive than that of the conventional PCR assay. The sensitivity of the LAMP assay in a spiked bovine vaginal mucus specimen was similar to that of the conventional PCR assay. The assay was markedly faster, requiring less than 40 min for detection of C. fetus in a single colony on blood agar and 80 min in spiked bovine specimens from the beginning of DNA extraction to final determination. Our LAMP assay is a simple and practical tool for detection of C. fetus regardless of subspecies.
ISSN:	0378-1135 1873-2542
DOI:	10.1016/j.vetmic.2008.11.018