Development of a reliable dual-gene amplification RT-PCR assay for the detection of Turkey Meningoencephalitis virus in Turkey brain tissues

► Main findings indicated that no TMEV genome changes occurred but the PCR detection sensitivities of the env and NS5 genes differed. ► The amplification sensitivity of the NS5-gene was 10-100 times more than the env-gene, when separate. ► The newly developed multiplex RT-PCR was similar in sensitivity to the NS5-gene and can be considered as a sustainable diagnostic assay, because it double validates the TMEV diagnosis.