Rapid and sensitive detection of canine parvovirus type 2 by recombinase polymerase amplification

Wang, Jianchang; Liu, Libing; Li, Ruiwen; Wang, Jinfeng; Qi, Fazhi; Yuan, Wanzhe

doi:10.1007/s00705-015-2738-y

Cited by 31 publications

(23 citation statements)

References 17 publications

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“…Several assays have been reported for the detection or quantitation of CPV DNA [12][13][14][15][16][17][18], including PCR, nested PCR, iiPCR, RPA, LAMP-ELISA, LAMP-LFD, LAMP, polymerase spiral reaction, and SYBR Green based real-time PCR, but none of these assays enable differentiation CPV antigenic types and CPV-like viruses (MEV and FPV). Several other assays have been reported for the detection and differentiation of type 2-based vaccines and field strains of CPV [19] or typing of three antigenic types of CPV [20] or CPV and MEV [21] assay and MGB probe real-time RT-PCR, but none of these assays enables simultaneous detection and differentiation of four antigenic types of CPV.…”

Section: Introductionmentioning

confidence: 99%

A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

Sun

Cheng

Lin

et al. 2018

Molecular and Cellular Probes

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Canine parvovirus (CPV) is an important pathogen in domestic dogs, and the original antigenic types CPV-2 and its variants, CPV-2a, 2b and 2c, are prevalent worldwide. A multiplex TaqMan real-time PCR method was developed for the detection and differentiation of four antigenic types of CPV. A set of primers and probes, CPV-305F/CPV-305R and CPV-2-305P (for CPV-2)/CPV-2a-305P (for CPV-2a, 2b and 2c), was able to differentiate CPV-2 and its variants (CPV-2a, 2b and 2c). Another set of primers and probes, CPV-426F/CPV-426R and CPV-2-426P (for CPV-2 and 2a)/CPV-2b-426P (for CPV-2b)/CPV-2c-426P (for CPV-2c), was able to differentiate CPV-2a (2), CPV-2b, and CPV-2c. With these primers and probes, the multiplex TaqMan real-time PCR assay detected effectively and differentiated CPV-2, 2a, 2b and 2c by two separate real-time PCRs. No cross reactivity was observed with canine distemper virus, canine adenovirus, and canine coronavirus. The detection limit of the assay is 10 genome copies/μL for CPV-2, CPV-2a, CPV-2b, and 10 copies/μL for CPV-2c. The multiplex real-time PCR has 100% agreement with DNA sequencing. We provide a sensitive assay that simultaneously detects and differentiate four antigenic types of CPV and the method was also used for quantification of CPVs viral genome.

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Section: Introductionmentioning

confidence: 99%

A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

Sun

Cheng

Lin

et al. 2018

Molecular and Cellular Probes

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“…RPA has been developed for veterinary importance viruses detection, including porcine parvovirus virus [13], avian influenza virus [14], bovine viral diarrhea virus [15], canine parvovirus type 2 [16], foot-and-mouth disease virus [17], and Orf virus [18, 19]. In this study, we developed fluorescent probe-based real-time RPA assay (PCV2 real-time RPA assay) and RPA assay united with LFD (PCV2 RPA LFD assay) to detect PCV2.…”

Section: Introductionmentioning

confidence: 99%

Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

Yang

Qin

Sun

et al. 2017

BioMed Research International

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Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 102 copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.

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“…In addition, the SNAP test based on ELISA protocols for detection of viral antigens is commonly used for CPV-2 diagnosis and can be completed in about 8 min by employing the commercial kit, whereas PCR seems to be more sensitive than SNAP [8,13,16,17]. Recently we reported on recombinase polymerase amplification (RPA) as a rapid, specific, sensitive and cost-effective molecular method for POC diagnosis of CPV-2 infection [18].…”

Section: Introductionmentioning

confidence: 99%

Development of real-time recombinase polymerase amplification assay for rapid and sensitive detection of canine parvovirus 2

Geng¹,

Wang²,

Liu³

et al. 2017

BMC Vet Res

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BackgroundCanine parvovirus 2, a linear single-stranded DNA virus belonging to the genus Parvovirus within the family Parvoviridae, is a highly contagious pathogen of domestic dogs and several wild canidae species. Early detection of canine parvovirus (CPV-2) is crucial to initiating appropriate outbreak control strategies. Recombinase polymerase amplification (RPA), a novel isothermal gene amplification technique, has been developed for the molecular detection of diverse pathogens. In this study, a real-time RPA assay was developed for the detection of CPV-2 using primers and an exo probe targeting the CPV-2 nucleocapsid protein gene.ResultsThe real-time RPA assay was performed successfully at 38 °C, and the results were obtained within 4–12 min for 105–101 molecules of template DNA. The assay only detected CPV-2, and did not show cross-detection of other viral pathogens, demonstrating a high level of specificity. The analytical sensitivity of the real-time RPA was 101 copies/reaction of a standard DNA template, which was 10 times more sensitive than the common RPA method. The clinical sensitivity of the real-time RPA assay matched 100% (n = 91) to the real-time PCR results.ConclusionThe real-time RPA assay is a simple, rapid, reliable and affordable method that can potentially be applied for the detection of CPV-2 in the research laboratory and point-of-care diagnosis.Electronic supplementary materialThe online version of this article (10.1186/s12917-017-1232-z) contains supplementary material, which is available to authorized users.

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Rapid and sensitive detection of canine parvovirus type 2 by recombinase polymerase amplification

Cited by 31 publications

References 17 publications

A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

A multiplex TaqMan real-time PCR for detection and differentiation of four antigenic types of canine parvovirus in China

Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2

Development of real-time recombinase polymerase amplification assay for rapid and sensitive detection of canine parvovirus 2

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