Characterisation of the canine parvovirus type 2 variants using minor groove binder probe technology.
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.
Decaro
Nicola
N
Department of Animal Health and Well-Being, Faculty of Veterinary Medicine of Bari, 70010 Valenzano, Bari, Italy. n.decaro@veterinaria.uniba.it
Elia
Gabriella
G
Martella
Vito
V
Campolo
Marco
M
Desario
Costantina
C
Camero
Michele
M
Cirone
Francesco
F
Lorusso
Eleonora
E
Lucente
Maria Stella
MS
Narcisi
Donato
D
Scalia
Pierluigi
P
Buonavoglia
Canio
C
D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004283:Dog Diseases / Q000235:genetics / Q000821:virology*; D004285:Dogs; D005243:Feces / Q000821:virology; D018628:Gene Dosage; D014644:Genetic Variation; D006385:Hemagglutination Inhibition Tests; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D017993:Parvovirus, Canine / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D019914:Taq Polymerase
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D015342:DNA Probes; D004279:DNA, Viral; D019914:Taq Polymerase