LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis

Brazilian Journal of Microbiology

Marcos D. Trangoni¹  Andrea K. Gioffré¹  María E. Cerón Cucchi¹  Karina C. Caimi¹  Paula Ruybal¹  Martín J. Zumárraga¹  Silvio L. Cravero¹ 

关键词: loop-mediated isothermal amplification;    molecular typing;    brucellosis;    paratuberculosis;   

DOI  :  10.1590/S1517-838246220131206

来源: SciELO

【 摘 要 】

In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.

【 授权许可】

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