Development of multiplex RT-PCR for detection and differentiation of foot-and-mouth disease virus O and A serotypes in Turkey

Authors : Beyhan SAREYYÜPOĞLU, İbrahim BURGU

Pages : 764-769

View : 15 | Download : 3

Publication Date : 0000-00-00

Article Type : Research Paper

Abstract :  In this study, a two-step multiplex reverse transcription-polymerase chain reaction insert ignore into journalissuearticles values(mRT-PCR); was developed and evaluated using designed primers for the simultaneous detection and differentiation of Turkish foot-and-mouth disease virus insert ignore into journalissuearticles values(FMDV); serotypes A and O from clinical samples. In the method, a mix of one universal reverse primer designed from the 2B gene and two serotype-specific forward primers designed from the hypervariable regions of the capsid VP1-1D coding gene of FMDV were used. Totally, 272 FMDV-infected samples collected between 2006 and 2008 constituted the material of the study. mRT-PCR was compared to ELISA in terms of positivity percentage insert ignore into journalissuearticles values(%); and the difference between the two tests was found to be statistically significant insert ignore into journalissuearticles values(P < 0.05);. In the study, 148 insert ignore into journalissuearticles values(54.4%); of 272 samples were undetermined by ELISA. Ninety-nine insert ignore into journalissuearticles values(66.8%); of these 148 undetermined samples were found FMDV RNA-positive with mRT-PCR. Consequently, the diagnostic sensitivity and specificity of mRT-PCR was determined as 95% and 84%, respectively. The study showed that mRT-PCR was more efficient than ELISA and concluded that the technique can be used as an alternative to ELISA for molecular typing of FMDV samples. This is the first report providing the differentiation of FMDV serotypes A and O in Turkey by mRT-PCR.
Keywords : Foot and mouth disease virus, diagnosis, mRT PC