Abstract
Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method offering rapid, accurate, and cost-effective diagnosis of infectious diseases. From the beginning of DNA extraction to final detection of Campylobacter jejuni and Campylobacter coli, the assay requires less than 50 and 90 min from a colony on selective media, and human feces, respectively. For chicken meat samples, the assay requires approximately 24–48 h from the beginning of the enrichment culture to final detection. The sensitivity of the LAMP assay is tenfold higher than that of the equivalent PCR assay. LAMP amplification can be judged by both turbidimeter analysis and visual assessment with the unaided eye. The LAMP assay is a powerful tool for rapid, simple, and sensitive detection of C. jejuni and C. coli, which may facilitate the investigation of C. jejuni and C. coli contamination in chicken, as well as the early diagnosis of C. jejuni and C. coli infection in humans.
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References
Yamazaki-Matsune W, Taguchi M, Seto K, Kawahara R, Kawatsu K, Kumeda Y, Kitazato M, Nukina M, Misawa N, Tsukamoto T (2007) Development of a multiplex PCR assay for identification of Campylobacter coli, Campylobacter fetus, Campylobacter hyointestinalis subsp. hyointestinalis, Campylobacter jejuni, Campylobacter lari and Campylobacter upsaliensis. J Med Microbiol 56:1467–1473
Logan JM, Edwards KJ, Saunders NA, Stanley J (2001) Rapid identification of Campylobacter spp. by melting peak analysis of biprobes in real-time PCR. J Clin Microbiol 39:2227–2232
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28:e63
Mori Y, Nagamine K, Tomita N, Notomi T (2001) Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 289:150–154
Nagamine K, Hase T, Notomi T (2002) Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16:223–229
Yamazaki W, Taguchi M, Ishibashi M, Kitazato M, Nukina M, Misawa N, Inoue K (2008) Development and evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Campylobacter jejuni and Campylobacter coli. J Med Microbiol 57:444–451
Yamazaki W, Taguchi M, Kawai T, Kawatsu K, Sakata J, Inoue K, Misawa N (2009) Comparison of loop-mediated isothermal amplification assay and conventional culture methods for detection of Campylobacter jejuni and Campylobacter coli in naturally contaminated chicken meat samples. Appl Environ Microbiol 75:1597–1603
Kaneko H, Kawana T, Fukushima E, Suzutani T (2007) Tolerance of loop-mediated isothermal amplification to a culture medium and biological substances. J Biochem Biophys Methods 70:499–501
Mohran ZS, Arthur RR, Oyofo BA, Peruski LF, Wasfy MO, Ismail TF, Murphy JR (1998) Differentiation of Campylobacter isolates on the basis of sensitivity to boiling in water as measured by PCR-detectable DNA. Appl Environ Microbiol 64:363–365
Wilson IG (1997) Inhibition and facilitation of nucleic acid amplification. Appl Environ Microbiol 63:3741–3751
Kanki M, Sakata J, Taguchi M, Kumeda Y, Ishibashi M, Kawai T, Kawatsu K, Yamasaki W, Inoue K, Miyahara M (2009) Effect of sample preparation and bacterial concentration on Salmonella enterica detection in poultry meat using culture methods and PCR assaying of preenrichment broths. Food Microbiol 26:1–3
Parkhill J, Wren BW, Mungall K, Ketley JM, Churcher C, Basham D, Chillingworth T, Davies RM, Feltwell T, Holroyd S, Jagels K, Karlyshev AV, Moule S, Pallen MJ, Penn CW, Quail MA, Rajandream MA, Rutherford KM, van Vliet AH, Whitehead S, Barrell BG (2000) The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature 403:665–668
Fouts DE, Mongodin EF, Mandrell RE, Miller WG, Rasko DA, Ravel J, Brinkac LM, DeBoy RT, Parker CT, Daugherty SC, Dodson RJ, Durkin AS, Madupu R, Sullivan SA, Shetty JU, Ayodeji MA, Shvartsbeyn A, Schatz MC, Badger JH, Fraser CM, Nelson KE (2005) Major structural differences and novel potential virulence mechanisms from the genomes of multiple Campylobacter species. PLoS Biol 3:e15
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Yamazaki, W. (2013). Sensitive and Rapid Detection of Campylobacter jejuni and Campylobacter coli Using Loop-Mediated Isothermal Amplification. In: Wilks, M. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology, vol 943. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-353-4_18
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DOI: https://doi.org/10.1007/978-1-60327-353-4_18
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Publisher Name: Humana Press, Totowa, NJ
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