Abstract
BackgroundWaterfowl parvoviruses, including goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV), can cause seriously diseases in geese and ducks. Developing a fast and precise diagnosis assay for these two parvoviruses is particularly important.ResultsA duplex SYBR Green I-based quantitative real-time PCR assay was developed for the simultaneous detection and differentiation of GPV and MDPV. The assay yielded melting curves with specific single peak (Tm = 87.3 ± 0.26 °C or Tm = 85.4 ± 0.23 °C) when GPV or MDPV was evaluated, respectively. When both parvoviruses were assessed in one reaction, melting curves with specific double peaks were yielded.ConclusionThis duplex quantitative RT-PCR can be used to rapid identify of GPV and MDPV in field cases and artificial trials, which make it a powerful tool for diagnosing, preventing and controlling waterfowl parvovirus infections.
Highlights
Waterfowl parvoviruses cause substantial economic loss to waterfowl production worldwide [9, 12]
Quantification method to detect waterfowl parvoviruses was previously reported using a TaqMan-based real-time polymerase chain reaction (PCR) reaction containing primers targeting the parvovirus, Lin et al Virology Journal (2019)6: as well as a probe labeled with fluorescent reporter dyes
The results indicated that 41 of the 65 artificial samples were infected with goose parvovirus (GPV), which had single peaks in the melting curves at Tm = 87.3 ± 0.32 °C and the other 24 samples were infected with Muscovy duck parvovirus (MDPV), resulting in single melting peaks at Tm = 85.2 ± 0.26 °C
Summary
Waterfowl parvoviruses cause substantial economic loss to waterfowl production worldwide [9, 12]. Quantification method to detect waterfowl parvoviruses was previously reported using a TaqMan-based real-time PCR reaction containing primers targeting the parvovirus, Lin et al Virology Journal (2019): as well as a probe labeled with fluorescent reporter dyes. These assays were not developed to differentiate GPV and MDPV [11, 16, 24, 28]. We developed a duplex SYBR Green-I based quantitative real-time PCR to rapidly and amplify a region of the GPV and MDPV genome that can be used to distinguish infection by either virus in a single reaction.
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