Journal Article

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

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

2019

Overview

Background Classic 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. Results After 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. Conclusions We 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.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: Analysis / Animals / Anseriform dependoparvovirus 1 / beak / breeding / Cairina moschata / China / Classic GPV / cross reaction / detection limit / Differentiation / DNA / DNA, Bacterial - genetics / DNA, Complementary - genetics / DNA, Viral - genetics / ducklings / Ducks / dwarfing / Dwarfism / epidemiological studies / Epidemiology / Escherichia coli / Fluorescence / Geese / Genes / goslings / Health aspects / Host Specificity / Medicine / Medicine & Public Health / Morbidity / Mortality / mules / Muscovy / N-GPV / Novels / NS gene / Parvoviridae Infections - diagnosis / Parvoviridae Infections - veterinary / Parvoviridae Infections - virology / Parvovirinae - isolation & purification / pathogenesis / pathogenicity / Pekin / Polymerase Chain Reaction - methods / Poultry Diseases - diagnosis / Poultry Diseases - virology / quantitative polymerase chain reaction / Real-Time Polymerase Chain Reaction / Reproducibility of Results / Research Article / Sensitivity and Specificity / Specific detection / TaqMan real-time PCR assay / Transgenics / Veterinary Medicine/Veterinary Science / Virology / Zoology