Simultaneous detection of duck circovirus and novel goose parvovirus via SYBR green I-based duplex real-time polymerase chain reaction analysis

Abstract

Beak atrophy and dwarfism syndrome (BADS) is commonly caused by co-infection with duck circovirus (DuCV) and novel goose parvovirus (NGPV). Therefore, concurrent detection of both viruses is important for monitoring and limiting BADS, although such a diagnostic test has not been reported. In this study, we developed a duplex, SYBR Green I-based real-time polymerase chain reaction (PCR) assay to enable the simultaneous detection of DuCV and NGPV. The assay readily distinguished between the two viruses, based on their different melting temperatures (Tm), where the Tm for DuCV was 80 °C and that for NGPV was 84.5 °C. Other non-target duck viruses that were tested did not show melting peaks. The detection limit of the duplex assay was 10 1 copies/μL for both viruses. This method exhibited high repeatability and reproducibility, and both the inter-assay and intra-assay variation coefficients were <1.6%. Thirty-one fecal samples were collected for clinical testing using real-time PCR analysis, and the results were confirmed using sequencing. The rate of co-infection was 6.5%, which was consistent with the sequencing results. This duplex real-time PCR assay offers advantages over other tests, such as rapid, sensitive, specific, and reliable detection of both viruses in a single sample, which enables the quantitative detection of DuCV and NGPV in clinical samples. Using this test may be instrumental in reducing the incidence of BADS and the associated economic losses in the duck and goose industries.

Reproduction and pathogenesis of short beak and dwarfish syndrome in Cherry Valley Pekin ducks infected with the rescued novel goose parvovirus

“…Since the outbreak of SBDS in China, NGPV was isolated from the clinical SBDS organ tissues [ 1 , 2 ]. However, NGPV co-infection with duck circovirus was also reported to be present in some field samples [ 16 , 17 ], hence, whether sole NGPV infection can reproduce all the typical SBDS symptoms in Cherry Valley Pekin ducks still confuses us. In the present study, through the reverse genetics technology, the infectious NGPV carrying the genetic marker was rescued and used in the infection and horizontal transmission test.…”

“…Until now, little is known about the pathogenesis of NGPV toward Cherry Valley Pekin ducks. Furthermore, due to the finding of duck circovirus co-infection in the SBDS clinical cases [ 16 , 17 ], whether sole NGPV infection in Cherry Valley Pekin duck can reproduce all the typical symptoms of SBDS remains unclear. In this study, based on the NGPV isolate SDJN19, an infectious plasmid clone pJNm containing the whole genome of SDJN19 was constructed.…”

Rapid diagnosis of duck Tembusu virus and goose astrovirus with TaqMan-based duplex real-time PCR

“…Against the most prevalent viral pathogens circulating in goose, like Goose parvovirus (GPV), Newcastle disease virus (NDV), goose herpesvirus (GHV), and goose adenovirus (GAV), several molecular assays have been developed for clinical diagnosis in the past few years by using either genomic detection methods such as PCR or antigen detection methods such as ELISA ( Chen et al, 2009 ; Guo et al, 2009 ; Wan et al, 2019 ; Wu et al, 2019 ; Tomar et al, 2021 ). Also, several duplex detection assays were developed for rapid screening of multiple pathogens, usually by duplex PCR ( Kong et al, 2007 ; Chen et al, 2013 ; Wang et al, 2020 ). These methods have brought great convenience and benefits to the breeding industry.…”