I’m really looking to find out the most sensitive and specific method for the diagnosis as I read different methods in which different viruses genes were detected.
If you are interested in sensitivity and specificity you should also take a look at dPCR protocols.
Hi,
We use one step real-time PCR method described by Batten et al. (2011) in our laboratory. In our experience, this protocol has a good sensitivity and specificity for detection of PPRV.
"Batten et al. (2011). A real time RT-PCR assay for the specific detection of Peste des petits ruminants virus. Journal of Virological Methods 171 (2011) 401–404."
Hi Wafa,
As for protocol for FMD viruses assessment by using one step Real-Time PCR method, please use the beneath publication of Laila EL-Shehawy et al. "Laboratory diagnosis of FMD using real-time RT-PCR in Egypt", published in Life Science Journal 8(3):384-387, 2011. Abstract: Definitive diagnosis of foot-and-mouth disease (FMD) requires the detection of virus antigen or genome in clinical material. The aim was performance of real-time RT-PCR (rRT-PCR) procedures for this purpose. Twenty nine cattle samples of vesicular epithelial and nasal swabs from four localities of Ismailia governorates were examined by ELISA, VN, RT-PCR and rRT-PCR. The results showed that 11 samples were positive by ELISA and virus isolation, 8 of serotype O and 3 for type A. Fourteen samples out of 29 were positive by RT-PCR and rRTPCR. The features that influence sample quality appear to be less important for the rRT-PCR and RT-PCR as they can detect a small fragment of FMDV genomic RNA. Real-time RT-PCR provided an extremely sensitizer and rapid procedures that contributes to improved laboratory diagnosis of FMD.
Best wishes,
Ilya
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