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

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.

Cabras

“…Las pruebas de diagnóstico serológico pueden practicarse a partir del tercer día posterior a la infección y primordialmente se realiza la prueba de inhibición de la hemoaglutinación, que se basa en la capacidad de la proteína HN de hemoaglutinar glóbulos rojos; también se han descrito pruebas como elisa, IF e ipma (Graham et al, 1998;Mao et al, 2017). Para amplificar un fragmento del genoma viral se efectúa la prueba de rt-pcr punto final y rt-pcr tiempo real, en las cuales se amplifican los genes HN y N (Templeton et al, 2004;Vaucher, Simonetti y Roehe, 2008). A B…”

A lateral flow dipstick combined with reverse transcription recombinase polymerase amplification for rapid and visual detection of the bovine respirovirus 3

“…The common and widely accepted molecular technique for detecting BPIV3 is reverse transcriptase polymerase chain reaction (RT-PCR) or quantitative RT-PCR (RT-qPCR) [6,7]. However, this method requires thermal cycling equipment and the settings of professional diagnostic laboratory.…”