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

Abstract

In this study, a two-step multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed and evaluated using designed primers for the simultaneous detection and differentiation of Turkish foot-and-mouth disease virus (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 (%) and the difference between the two tests was found to be statistically significant (P < 0.05). In the study, 148 (54.4%) of 272 samples were undetermined by ELISA. Ninety-nine (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.