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

TRANGONI, MARCOS DAVID; GIOFFRÉ, ANDREA KARINA; CERÓN CUCCHI, MARÍA; CAIMI, KARINA; RUYBAL P; ZUMÁRRAGA, MARTÍN JOSÉ; CRAVERO, SILVIO

BRAZILIAN JOURNAL OF MICROBIOLOGY

SOC BRASILEIRA MICROBIOLOGIA

Año: 2015 vol. 46

1517-8382

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-characterizedtarget gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection usingthe loop-mediated isothermal amplification (LAMP) protocols described herein were similar tothose of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTMallowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We includedthe LAMP-based protocol in a rapid identification scheme of the respective pathogens, and alltested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable forspecifically identifying the respective pathogens; in the case of Brucella, it also allowed the identificationof all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool thatcould help to shorten the time required to identify insidious bacteria in low-complexity laboratories,mainly in developing countries.