Specific detection and differentiation of classic goose parvovirus and novel goose parvovirus by TaqMan real-time PCR assay, coupled with host specificity

Abstract

BackgroundClassic goose parvovirus (cGPV) causes high mortality and morbidity in goslings and Muscovy ducklings. Novel GPV (N-GPV) causes short beak and dwarfism syndrome (SBDS) in Cherry Valley ducks, Pekin ducks and Mule ducks. Both cGPV and N-GPV have relatively strict host specificity, with obvious differences in pathogenicity. Specific detection of cGPV and N-GPV may result in false positives due to high nucleotide similarity with Muscovy duck parvovirus (MDPV). The aim of this study was to develop a highly specific, sensitive, and reliable TaqMan real-time PCR (TaqMan qPCR) assay for facilitating the molecular detection of cGPV and N-GPV.ResultsAfter genetic comparison, the specific conserved region (located on the NS gene) of cGPV and N-GPV was selected for primer and probe design. The selected regions were significantly different from MDPV. Through a series of optimization experiments, the limit of detection was 50.2 copies/μl. The assay was highly specific for the detection of cGPV and N-GPV and no cross-reactivity was observed with E. coli., P.M., R.A., S.S., MDPV, N-MDPV, DAdV-A, DEV, GHPV, DHAV-1, DHAV-3, ATmV, AIV, MDRV and N-DRV. The assay was reproducible with an intra-assay and inter-assay variability of less than 2.37%. Combined with host specificity, the developed TaqMan qPCR can be used for cGPV and N-GPV in differential diagnoses. The frequency of cGPV in Muscovy duckling and goslings was determined to be 12 to 44%, while N-GPV frequency in Mule ducks and Cherry Valley ducks was 36 to 56%. Additionally, fluorescence-positive signals can be found in Mule duck embryos and newly hatched Mule ducklings. These findings provide evidence of possible vertical transmission of N-GPV from breeding Mule ducks to ducklings.ConclusionsWe established a quantitative platform for epidemiological investigations and pathogenesis studies of cGPV and N-GPV DNA that was highly sensitive, specific, and reproducible. N-GPV and cGPV infections can be distinguished based on host specificity.

Detection of Novel Goose Parvovirus Disease Associated with Short Beak and Dwarfism Syndrome in Commercial Ducks

“…Earlier studies reported the possible vertical transmission of classic GPV in geese [ 19 ] and for MDPV [ 20 ]. Similarly, vertical transmission of NGPV from breeder cherry valley, pekin and mule ducks to their offspring was evidenced [ 17 , 21 , 22 ].…”

Molecular analysis of goose parvovirus field strains from the Derzsy’s disease outbreak revealed the local European-associated variants

“…Zadori et al (1995) previously de ned a primer set for the VP3 position [8], which has been extensively used to identify and classify GPV elsewhere [6,30,31]. Since then, additional methods have been developed such as Real Time PCR [32], LAMP [33] and immunochromatographic assay [34]. We de ned a novel degenerated primer set for the detection of GPV amplifying VP3 coding sequence.…”

Development of a sandwich enzyme-linked immunosorbent assay based on single-domain antibody for detecting goose parvovirus infection

“…The traditional methods used to detect GPV infection include detecting the GPV antigen or antibodies against GPV. Among GPV antigen detection methods, electron microscopy observations, polymerase chain reaction (PCR), real-time PCR, matrix-assisted laser desorption/ionization-time of flight mass spectrometry, loop-mediated isothermal amplification, and recombinant polymerase amplification combined with the vertical flow have been established to detect GPV (Wan et al, 2019). However, these techniques are more appropriate for laboratory analysis and are not relevant for field practice, as they are time-consuming methods that require specialized systems and trained personnel.…”