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

Creative Commons License

DİNÇER E., ÖZKUL A.

ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI, cilt.62, sa.4, ss.283-287, 2015 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 62 Sayı: 4
  • Basım Tarihi: 2015
  • Doi Numarası: 10.1501/vetfak_0000002693
  • Dergi Adı: ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.283-287
  • Anahtar Kelimeler: Peste des petits ruminants, Real-time qRT-PCR, virus detection, ASSAY
  • Ankara Üniversitesi Adresli: Evet

Özet

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.