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Evaluation of a duplex reverse-transcription real-time PCR assay for the detection of encephalomyocarditis virus

Qin, S., Underwood, D., Driver, L., Kistler, C., Diallo, I. and Kirkland, P. D. (2018) Evaluation of a duplex reverse-transcription real-time PCR assay for the detection of encephalomyocarditis virus. Journal of Veterinary Diagnostic Investigation, 30 (4). pp. 554-559.

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Article Link(s): https://doi.org/10.1177/1040638718779112

Publisher URL: http://journals.sagepub.com/doi/abs/10.1177/1040638718779112

Abstract

We evaluated a fluorogenic probe–based assay for the detection of encephalomyocarditis virus (EMCV) by comparing a set of published primers and probe to a new set of primers and probe. The published reagents failed to amplify a range of Australian isolates and an Italian reference strain of EMCV. In contrast, an assay based on 2 new sets of primers and probes that were run in a duplex reverse-transcription real-time PCR (RT-rtPCR) worked well, with high amplification efficiency. The analytical sensitivity was ~100-fold higher than virus isolation in cell culture. The intra-assay variation was 0.21–4.90%. No cross-reactivity was observed with a range of other porcine viruses. One hundred and twenty-two clinical specimens were tested simultaneously by RT-rtPCR and virus isolation in cell culture; 72 specimens gave positive results by RT-rtPCR, and 63 of these were also positive by virus isolation. Of 245 archived cell culture isolates of EMCV that were tested in the RT-rtPCR, 242 samples were positive. The new duplex RT-rtPCR assay is a reliable tool for the detection of EMCV in clinical specimens and for use in epidemiologic investigations.

Item Type:Article
Business groups:Biosecurity Queensland
Keywords:Encephalomyocarditis virus,reverse-transcription polymerase chain reaction
Subjects:Veterinary medicine
Veterinary medicine > Veterinary virology
Deposited On:14 Aug 2018 04:36
Last Modified:14 Aug 2018 04:36

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