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      A minor groove binder probe real-time PCR assay for discrimination between type 2-based vaccines and field strains of canine parvovirus

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          Abstract

          A minor groove binder (MGB) probe assay was developed to discriminate between type 2-based vaccines and field strains of canine parvovirus (CPV). Considering that most of the CPV vaccines contain the old type 2, no longer circulating in canine population, two MGB probes specific for CPV-2 and the antigenic variants (types 2a, 2b and 2c), respectively, were labeled with different fluorophores. The MGB probe assay was able to discriminate correctly between the old type and the variants, with a detection limit of 10 1 DNA copies and a good reproducibility. Quantitation of the viral DNA loads was accurate, as demonstrated by comparing the CPV DNA titres to those calculated by means of the TaqMan assay recognising all CPV types. This assay will ensure resolution of most diagnostic problems in dogs showing CPV disease shortly after CPV vaccination, although it does not discriminate between field strains and type 2b-based vaccines, recently licensed to market in some countries.

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          Most cited references26

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          A real-time PCR assay for rapid detection and quantitation of canine parvovirus type 2 in the feces of dogs.

          We describe a rapid, sensitive and reproducible real-time PCR assay for detecting and quantifying canine parvovirus type 2 (CPV-2) DNA in the feces of dogs with diarrhea. An exogenous internal control was added to control the assay performance from extraction to amplification. The method was demonstrated to be highly specific and sensitive, allowing a precise CPV-2 DNA quantitation over a range of eight orders of magnitude (from 10(2) to 10(9) copies of standard DNA). The reproducibility of the CPV-2 real-time PCR assay was assessed by calculating the coefficients of variation (CV) intra-assay and inter-assay for samples containing amounts of CPV-2 DNA spanning the whole range of the real-time PCR standard curve. Then, fecal specimens from diarrheic dogs were analyzed by hemagglutination (HA), conventional PCR and real-time amplification. Comparison between these different techniques revealed that real-time PCR is more sensitive than HA and conventional gel-based PCR, allowing to detect low viral titers of CPV-2 in infected dogs.
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            Natural variation of canine parvovirus.

            Canine parvovirus was first recognized during 1978. Analysis of isolates collected since its emergence revealed that viruses circulating after 1980 were antigenically different from earlier isolates. Monoclonal antibodies clearly distinguished the two strains, some being specific for either the old or the new viruses. Restriction enzyme analysis of viral DNA's showed that the post-1980 viruses were similar to earlier isolates, but some restriction site differences were present in the new strain. These results suggest that the canine parvoviruses infecting dogs in the seven areas of the United States that were sampled derive from a variant virus that replaced the original strain during 1980.
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              A novel antigenic variant of Canine parvovirus from a Vietnamese dog.

              Nine isolates of Canine parvovirus (CPV) were obtained from Vietnamese dogs and cats. One canine isolate showed a unique antigenic property which indicates a novel antigenic variant of CPV-2b when examined with hemagglutination inhibition tests using our monoclonal antibodies, 21C3 and 19D7, which were recently developed. This isolate had an amino acid substitution of residue 426, Asp to Glu, and the same substitution has recently been found in CPV from Italian dogs. This study first showed that such substitution caused an antigenic difference demonstrable by monoclonal antibodies and that a similar evolution may have occurred in CPV in Vietnam.
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                Author and article information

                Contributors
                Journal
                J Virol Methods
                J. Virol. Methods
                Journal of Virological Methods
                Elsevier B.V.
                0166-0934
                1879-0984
                6 May 2006
                September 2006
                6 May 2006
                : 136
                : 1
                : 65-70
                Affiliations
                [a ]Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, Strada per Casamassima Km 3, 70010 Valenzano (Bari), Italy
                [b ]Department of Pathology and Animal Health, Faculty of Veterinary Medicine of Naples, via Delpino 1, 80137 Naples, Italy
                Author notes
                [* ]Corresponding author. Tel.: +39 080 4679832; fax: +39 080 4679843. n.decaro@ 123456veterinaria.uniba.it
                Article
                S0166-0934(06)00124-8
                10.1016/j.jviromet.2006.03.030
                7119799
                16682086
                640617c7-3d4f-4e86-a7a0-648455320fd1
                Copyright © 2006 Elsevier B.V. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 27 December 2005
                : 28 March 2006
                : 30 March 2006
                Categories
                Article

                Microbiology & Virology
                dog,parvovirus,type 2 vaccine,field strains,mgb probe assay
                Microbiology & Virology
                dog, parvovirus, type 2 vaccine, field strains, mgb probe assay

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