VALIDATION OF A RT-qPCR ASSAY FOR DIFFERENTIAL DIAGNOSIS OF AFRICAN AND CLASSICAL SWINE FEVER
DOI:
https://doi.org/10.31548/dopovidi2017.06.014Keywords:
African swine fever (ASF), Classical swine fever (CSF), RT-qPCR, validation.Abstract
A real-time reverse-transcriptase polymerase chain reaction (RT-qPCR) assay for differential diagnosis of African and Classical swine fever has been developed in the Institute of Veterinary Medicine of the NAAS. The proposed assay allows simultaneous detection of three targets: ASFV, CSFV and an internal control, which helps to save time and money. In this article, an inter-laboratory validation of the developed RT-qPCR assay was described. Validation of the test kit was performed in compliance with OIE Validation Guidelines according to analytical sensitivity, specificity and repeatability. The limit of detection (LOD) of the validated RT-qPCR assay was 10 copies of the ASFV and CSFV genomes, respectively. Repeatability of the assay was shown to be high enough. The specificity of the assay, evaluated on different viral strains showed no cross-reactions with closely related pathogens (porcine circovirus type 2, porcine reproductive and respiratory syndrome virus and virus of Aujeszky's disease). The expected reactivity for ASFV and CSFV were observed. In conclusion, all investigated analytical performance criteria of the validated RT-qPCR assay for differential diagnosis of ASF and CSF are in compliance with international standards, which ensures accurate and definite results. This laboratory assay will be a valuable tool for rapid differential diagnosis in case of swine fever suspected outbreaks.
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