Since 2013, outbreaks of disease caused by duck Tembusu virus (DTMUV) have been observed in layer and broiler duck farms in Thailand. The virus is closely related to Chinese DTMUVs and belongs to the Ntaya group of mosquitoborne flaviviruses. These findings represent the emergence of DTMUV in ducks in Thailand.
Aims: To develop and validate assays based on real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR) for rapid detection and strain identification (European and North American strains) of porcine reproductive and respiratory syndrome virus (PRRSV) by using SYBR Green I and TaqMan probe chemistries.
Methods and Results: This study describes two alternative assays based on real‐time RT‐PCR for rapid detection and strain identification of PRRSV in comparison with conventional RT‐PCR. The first assay utilized SYBR Green I with melting curve analysis; another assay was performed using strain‐specific TaqMan probes. Primers were selected from the conserved regions within ORF7 (N) of both strains whereas two TaqMan probes labelled with different fluorescent dyes were specifically designed for each strain. The result of strain identification was confirmed by direct sequencing. Both assays can be used for rapid detection and strain identification of PRRSV with a sensitivity of 104 and 103 copies μl−1 for SYBR Green and TaqMan probe, respectively.
Conclusions: Real‐time RT‐PCR is a powerful method combining rapidity, specificity and efficiency for large‐scale screening and strain identification of PRRSV.
Significance and Impact of the Study: The data indicate that the methods developed are invaluable for detecting low levels of PRRSV infection in swine.
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