Specific detection of Muscovy duck parvovirus infection by TaqMan-based real-time PCR assay

Chunhe Wan,Qiuling Fu,Yu Huang,Hongmei Chen,Guanghua Fu,Shaohua Shi,Rongchang Liu,Longfei Cheng,Cuiteng Chen

doi:10.1186/s12917-018-1600-3

Chunhe Wan, Qiuling Fu + Show 7 more

Open Access

https://doi.org/10.1186/s12917-018-1600-3

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Abstract

BackgroundMuscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic similarity with goose parvovirus (GPV). The objective of this study was to develop a sensitive, highly specific, and repeatable TaqMan-based real-time PCR (qPCR) assay for facilitating the molecular detection of MDPV.ResultsThe specific primers and probe were designed based on the conserved regions within MDPVs, but there was a variation in GPVs of the nonstructural (NS) genes after genetic comparison. After the optimization of qPCR conditions, the detection limit of this qPCR assay was 29.7 copies/μl. The assay was highly specific for the detection of MDPV, and no cross-reactivity was observed with other non-targeted duck-derived pathogens. Intra- and inter-assay variability was less than 2.21%, means a high degree of repeatability. The diagnostic applicability of the qPCR assay was proven that MDPV-positive can be found in cloacal swabs samples, Muscovy duck embryos and newly hatched Muscovy ducklings.ConclusionsOur data provided incidents that MDPV could be possible vertically transmitted from breeder Muscovy ducks to Muscovy ducklings. The developed qPCR assay in the study could be a reliable and specific tool for epidemiological surveillance and pathogenesis studies of MDPV.

Highlights

Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects
A terminal resolving site (TRS), Rep protein binding site (RBS), and transcription factor binding sites can be found in inverted terminal repeats (ITR), which were involved in viral replication, packaging and transcription
NS gene analysis We compared a total of 52 NS gene sequences downloaded from the GenBank database

Summary

Introduction

Muscovy duck parvovirus (MDPV) causes high mortality and morbidity in Muscovy ducks, with the pathogenesis of the virus still unknown in many respects. Specific MDPV detection is often rife with false positive results because of high identity at the genomic nucleotide level and antigenic similarity with goose parvovirus (GPV). Dependoparvovirus in subfamily Parvovirinae due to similar genetic properties and evolutionary origins (https://talk.ictvonline.org/taxonomy/). The genomes of MDPVs and GPVs contain a single copy of the linear, single-stranded DNA genome of approximately 5100 nucleotides in length. The genomes of these viruses are flanked by identical inverted terminal repeats (ITR) at both the 5′- and 3′-terminus. A terminal resolving site (TRS), Rep protein binding site (RBS), and transcription factor binding sites can be found in ITR, which were involved in viral replication, packaging and transcription. The left ORF that encodes for the nonstructural (NS) protein, which is involved in viral replication and

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