RT-PCR for detection of bovine parainfluenza virus type 3 (bPIV-3)

Authors

  • Rodrigo de Almeida Vaucher UFRGS
  • Amauri Braga Simonetti UFRGS
  • Paulo Michel Roehe UFRGS

DOI:

https://doi.org/10.22456/1679-9216.17288

Keywords:

Vírus parainfluenza tipo 3, bPIV-3, TR-PCR, gene da hemaglitinina-neuraminidase (HN), Detecção

Abstract

The RT-PCR technique has been frequentely used for detection of the human parainfluenza virus type 3 (hPIV-3) but the literature is scarce in relation to the bovine parainfluenza virus type 3 (bPIV-3). The aim of this study was to describe a reverse transcriptase polymerase chain reaction (RT-PCR) for detection of bovine parainfluenza virus type 3 (bPIV-3) using degenerate oligonucleotides targeting a conserved region of hemagglutinin-neuraminidase (HN) gene. Reference strain SF-4 and three different brazilian bPIV-3 isolates, besides five viral strains from different sources, were included in this study. Viruses were cultured in MDBK cells under standard conditions. Hemagglutination (HA) test was used for viral titration and a direct immunofluorescence test (DFAT) for isolate screening. In RT-PCR all bPIV-3 isolates showed amplification of an expected 1009 bp fragment of HN gene, as oposed to non PIV-3 viral samples where no amplification was detected. Using SF-4 as positive control, sensitivity of 95 pg cDNA wasachieved. In spite of the low number of bPIV-3 isolates tested, the results obtained in this study point out the potential use of this technique for detection of bPIV-3 in bovine clinical specimens.

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Author Biographies

Rodrigo de Almeida Vaucher, UFRGS

Amauri Braga Simonetti, UFRGS

 

 

Departamento de Microbiologia, Instituto de Ciências Básicas da Saúde (ICBS-UFRGS), Porto Alegre, RS/Brasil.

Paulo Michel Roehe, UFRGS

 

 

Departamento de Microbiologia, Instituto de Ciências Básicas da Saúde (ICBS-UFRGS), Porto Alegre, RS/Brasil.

 

 

Instituto de Pesquisas Veterinárias Desidério Finamor (IPVDF-FEPAGRO), Eldorado do Sul, RS/Brasil.

Published

2018-03-30

How to Cite

Vaucher, R. de A., Simonetti, A. B., & Roehe, P. M. (2018). RT-PCR for detection of bovine parainfluenza virus type 3 (bPIV-3). Acta Scientiae Veterinariae, 36(3), 215–220. https://doi.org/10.22456/1679-9216.17288

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