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J Virol Methods. 2006 Apr;133(1):92-9. Epub 2005 Nov 28.

Characterisation of the canine parvovirus type 2 variants using minor groove binder probe technology.

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  • 1Department of Animal Health and Well-Being, Faculty of Veterinary Medicine of Bari, 70010 Valenzano, Bari, Italy. n.decaro@veterinaria.uniba.it

Abstract

Two minor groove binder (MGB) probe assays were developed for rapid identification and quantitation of the canine parvovirus type 2 (CPV-2) variants in the faecal samples of dogs. The MGB probe assays using type-specific probes labeled with different fluorophores (FAM and VIC) were able to detect the single nucleotide polymorphisms existing between types 2a/2b and 2b/2c. Both the MGB probe assays were found to be highly specific, sensitive and reproducible, ensuring a precise quantitation of the CPV-2 DNA, as confirmed by comparing the obtained viral DNA loads to those calculated using a real-time TaqMan-based PCR assay established previously. A total of 414 faecal specimens collected from diarrhoeic dogs and tested positive to CPV-2 were analysed by the MGB probe assays and by conventional methods (haemagglutination inhibition with MAbs, PCR-RFLP and sequence analysis) for prediction of the CPV-2 antigen specificity, with an overall agreement of 100%. These results make the established MGB probe assays an attractive tool for direct and rapid characterisation of type 2a, 2b and 2c CPVs.

PMID:
16313976
[PubMed - indexed for MEDLINE]
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