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

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

doi:10.1186/s12917-018-1600-3

Cited by 20 publications

(12 citation statements)

References 34 publications

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“…Similarly, there was possible vertical transmission of N-GPV between breeder Cherry Valley and Pekin ducks to their ducklings [22, 23]. Classic MDPV shared the same phenomenon of possible vertical transmission, similar to our recent work [18]. In this study, we demonstrated that N-GPV appeared to possible vertically transfer from breeder Mule ducks to ducklings.…”

Section: Discussionsupporting

confidence: 86%

“…In this study, a total of 52 NS gene sequences (37 GPVs and 15 MDPVs) were compared for primer and probe design. Previous studies showed that NS genes shared characteristic variations between GPVs and MDPVs that could be used to design more precise primers and probes [18, 19]. Using a similar strategy, a TaqMan real-time PCR for the detection and quantification of GPV was developed and evaluated.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies demonstrated that the NS gene homology between GPVs (cGPVs and N-GPVs) and MDPVs ranged from 80.8 to 83.4% and can be used for GPVs and MDPVs differentiation [18, 19]. After a bioinformatics analysis of the NS genes of GPVs (cGPVs and N-GPVs) and MDPVs specific primers and a probe were designed using Primer Premier Software version 5.0 (Premier Biosoft, Palo Alto, CA, USA) following a similar strategy that we used to develop a specific TaqMan-based real-time PCR for MDPV.…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2019

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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.

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Section: Discussionsupporting

confidence: 86%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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

et al. 2019

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“…Quantitative real-time PCR is now widely used in clinical settings. Compared with loop-mediated isothermal amplification, enzyme-linked immunosorbent assay, and indirect immunofluorescence assay, quantitative real-time PCR has advantages with respect to time and sensitivity [ [27] , [28] , [29] , [30] ]. Quantitative real-time PCR also has higher sensitivity, specificity, and positive detection rates than traditional PCR.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a duplex SYBR green I-based real-time polymerase chain reaction assay for the rapid detection of canine circovirus and canine astrovirus

Wang

Cui

et al. 2020

Molecular and Cellular Probes

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The similar clinical characteristics of canine circovirus (CaCV) and canine astrovirus (CaAstV) infections and high frequency of co-infection make diagnosis difficult. In this study, a duplex SYBR Green I-based real-time polymerase chain reaction (PCR) assay was established for the rapid, simultaneous detection of CaCV and CaAstV. Two pairs of specific primers were designed based on the Rep gene of CaCV and the Cap gene of CaAstV. By using the real-time PCR assay method, the two viruses can be distinguished by the difference in melting temperatures, 79 °C and 86 °C for CaCV and CaAstV, respectively. This assay had high specificity, showing no cross-reaction with other common canine viruses, as well as high sensitivity, with minimum detection limits of 9.25 × 10 1 copies/μL and 6.15 × 10 1 copies/μL for CaCV and CaAstV, respectively. Based on the mean coefficient of variation, the method had good reproducibility and reliability. In a clinical test of 57 fecal samples, the rates of positive detection by real-time PCR were 14.04% (8/57) and 12.28% (7/57) for CaCV and CaAstV, respectively, and the rate of co-infection was 8.77% (5/57). In conclusion, the newly established duplex SYBR Green I-based real-time PCR assay is sensitive, specific, reliable, and rapid and is an effective tool for the detection of co-infections with CaCV and CaAstV.

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“…In checking to see the intra-batch repeatability, triplicates of each dilution were detected, and according to the formula of the geometric mean Cq values / standard deviation calculated the coefficients of variation (CV). The coefficient of variation for inter-assay repeatability shows the differences among the measures at different times [33]. Three repeats were performed for each of the inter and intra assay analysis.…”

Section: Repeatability Analysis Of the Rt-qpcr Assaymentioning

confidence: 99%

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

Zhang

Liu

et al. 2020

BMC Vet Res

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Background: In China, Newly emerging duck reovirus (NDRV) variants have been causing major disease problems in cherry valley ducks. NDRV has the potential to cause high morbidity and 5-50% mortality rates. Severe hemorrhagic-necrosis in the liver and spleen were commonly seen in NDRV affected ducks. The availability of upgraded methods for rapid diagnosis of newly emerging DRV variants is crucial for successful DRV infection control and prevention. Results: In this study, we present a TaqMan-based real-time PCR assay (RT-qPCR) for the detection of NDRV infection. Using the conserved regions within the NDRV genome, we designed the specific primers and probe. The lower limit of detection for NDRV infection was 10 copies/μL (Ct values: 38.3) after the optimization of the RT-qPCR conditions. By cross-checking with other duck viral pathogens, no crossreactivity was observed confirming the assay was highly specific for the detection of NDRV. Reproducibility of the RT-qPCR was confirmed by intra-and inter-assay variability was less than 2.91%(Intra-assay variability of Ct values: 0.07-1.48%; Interassay variability of Ct values: 0.49-2.91%). This RT-qPCR and conventional PCR (cPCR) detected one hundred and twenty samples of NDRV infection from different regions. The result shows that the positive rates were 94.17 and 84.17% respectively. The detection rate of RT-qPCR rapid detection assay was 10% higher than that of the cPCR method. Conclusion: This research developed a highly sensitive, specific, reproducible and versatile of RT-qPCR for quantitatively detecting NDRV. It can be used to study the pathogenesis and epidemiology investigation of NDRV.

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

Cited by 20 publications

References 34 publications

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

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

Establishment of a duplex SYBR green I-based real-time polymerase chain reaction assay for the rapid detection of canine circovirus and canine astrovirus

A TaqMan-based real-time PCR assay for specific detection of novel duck reovirus in China

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