Tae-Hoon Kim
Kyung Hee University, South Korea
Title: Rapid multiplex molecular detection of Brucella genus, B. abortus, B. melitensis and B. suis using ultra-fast convection palm PCR
Biography
Biography: Tae-Hoon Kim
Abstract
Rapid detection and timely treatment of brucellosis is important for increasing the curative ratio, to prevent disease spread among animals and to reduce the risk of transfer to humans. In this study, we developed a rapid molecular diagnostics and differentiation method for Brucella species using convection Palm polymerase chain reaction (PCR). Three Brucella genus representatives, B. abortus, B. melitensis and B. suis, were successfully identified and differentiated. A common primer, IS711, specific to all Brucella species tested in this study and species-specific primers were used. When these primer sets were used in convection PCR, each species-specific primer set specifically detected the target species. Multiplex detection was performed with a mixture of the common forward and all reverse primers and three distinct species-specific DNA amplicons were unambiguously detected. Sensitivity of detection was also tested and tens or hundreds of copies, depending on the species were detected using this PCR approach and gel electrophoresis. The PCR reaction time could be reduced to 20 min with distinct, detectable DNA amplicons. These results suggest that the ultra-fast multiplex detection of Brucella species developed in this study may be useful in rapid field applications.