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

Rodrigo De Almeida Vaucher,Paulo Michel Roehe,Amauri Braga Simonetti

doi:10.22456/1679-9216.17288

Rodrigo De Almeida Vaucher, Paulo Michel Roehe + Show 1 more

Open Access

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

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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.

Highlights

Bovine parainfluenza virus type 3is a Respirovirus, member of Paramyxoviridae family, antigenically related to human parainfluenza virus type 3 [7]
The aim of the present study was to develop a reverse transcriptase polymerase chain reaction (RT-PCR) for detection of bovine parainfluenza virus type 3 (bPIV-3) and evaluate the test on three Brazilian bPIV-3 isolates
The RT-PCR described here, using primers representing part of bPIV-3 hemagglutinin-neuraminidase gene (HN), amplified a fragment of approximately 1009 bp with material extracted from SF-4

Summary

Introduction

Is a Respirovirus, member of Paramyxoviridae family, antigenically related to human parainfluenza virus type 3 (hPIV-3) [7]. It is an important pathogen for animals but can cause disease in humans. In cattle it is associated to the clinical picture known as “shipping fever’’ [13]. Most cases of respiratory infection with bPIV-3 occur with mild clinical signs. More severe infections may be associated with bacterial or virus agents [15]. BPIV-3 has been associated to abortion in cattle [18]. Laboratory detection of bPIV-3 infections is classically performed by virus isolation in cell culture (gold standard technique). Confirmation of the identity of the virus is usually achieved by HA [4,14]

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