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Year 2013 - Volume 33, Number 10
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Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP), 33(10):1222-1226
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ABSTRACT.- Zhang H., Zhu L., Zhou Y.C., Ji H.W., Dai H.B., Guo W.Z. & Xu Z.W. 2013. Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP). Pesquisa Veterinária Brasileira 33(10):1222-1226. Key Laboratory of Animal Biotechnology Center of Sichuan Province, College of Veterinary Medicine of Sichuan Agricultural University, Ya’an, Sichuan, 625014, PR China. E-mail: abtcxzw@126.com
Bordetella bronchiseptica causes acute and chronic respiratory infections in diverse animal species and occasionally in humans. In this study, we described the establishment of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP) assay for the detection of B. bronchiseptica. A set of primers towards a 235 bp region within the flagellum gene of B. bronchiseptica was designed with online software.. The specificity of the LAMP assay was examined by using 6 porcine pathogens and 100 nasal swabs collected from healthy pigs and suspect infected pigs. The results indicated that positive reactions were confirmed for all B. bronchiseptica and no cross-reactivity was observed from other non-B. bronchiseptica. In sensitivity evaluations, the technique successfully detected a serial dilutions of extracted B. bronchiseptica DNA with a detection limit of 9 copies, which was 10 times more sensitive than that of PCR. Compared with conventional PCR, the higher sensitivity of LAMP method and no need for the complex instrumentation make this LAMP assay a promising alternative for the diagnosis of B. bronchiseptica in rural areas and developing countries where there lacks of complex laboratory services. |
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