Development of real-time reverse transcriptase-polymerase chain reaction (rt RT-PCR) targeting four genes of peste des petits ruminants virus


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Dincer E., ÖZKUL A.

ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI, vol.62, no.4, pp.283-287, 2015 (SCI-Expanded) identifier

  • Publication Type: Article / Article
  • Volume: 62 Issue: 4
  • Publication Date: 2015
  • Doi Number: 10.1501/vetfak_0000002693
  • Journal Name: ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Page Numbers: pp.283-287
  • Keywords: Peste des petits ruminants, Real-time qRT-PCR, virus detection, ASSAY
  • Dokuz Eylül University Affiliated: No

Abstract

In this study, one-step real-time quantitative reverse transcription PCR (qRT-PCR) was developed for the first time and evaluated for detection of peste des petits ruminants virus (PPRV) in field samples obtained from distinct geographical areas of Turkey. Primers and probes targeting four PPRV genes (namely L, M, N and P) were designed based on full genome sequence (AJ849636) of the local isolate (Tu00). The detection limits of the assays were found to be 7 copies/mu L RNA for L, 6 copies/mu L RNA for P, 10 copies/mu L RNA for M and 700 copies/mu L RNA for N, respectively. Besides, the detection ratio for PPRV in 45 field samples was 100% (45) for L, 88.9% (40) for M, 77.8% (35) for P and 22.3% (10) for N, respectively. In conclution, one-step real-time quantitative reverse transcription PCR (qRT-PCR) assay reported here for L gene design provides rapid, specific and sensitive detection of PPRV in tissue samples obtained from field cases.