Identification of feline polycystic kidney disease mutation using fret probes and melting curve analysis

We developed and validated a real-time polymerase chain reaction (PCR) assay using fluorescent hybridization probes and melting curve analysis to identify the PKD1 exon 29 (C → A) mutation, which is implicated in polycystic kidney disease of cats. DNA was isolated from peripheral blood of 20 Persian...

Full description

Saved in:
Bibliographic Details
Published in:Research in veterinary science 2009-02, Vol.86 (1), p.88-90
Main Authors: Criado-Fornelio, A., Buling, A., Barba-Carretero, J.C.
Format: Article
Language:English
Subjects:
Agreements
Animals
Cat Diseases - diagnosis
Cat Diseases - genetics
Cats
Deoxyribonucleic acid
Disease
DNA
DNA - chemistry
DNA - genetics
FRET probes
Genetic test
Genetic testing
Melting
Microbiology
Mutation
Point Mutation
Polycystic kidney disease
Polycystic Kidney Diseases - diagnosis
Polycystic Kidney Diseases - genetics
Polycystic Kidney Diseases - veterinary
Polymerase Chain Reaction - veterinary
Polymorphism, Restriction Fragment Length
Real-time PCR
Temperature
Thermal cycling
Veterinary medicine
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:We developed and validated a real-time polymerase chain reaction (PCR) assay using fluorescent hybridization probes and melting curve analysis to identify the PKD1 exon 29 (C → A) mutation, which is implicated in polycystic kidney disease of cats. DNA was isolated from peripheral blood of 20 Persian cats. The employ of the new real-time PCR and melting curve analysis in these samples indicated that 13 cats (65%) were wild type homozygotes and seven cats (35%) were heterozygotes. Both PCR–RFLP and sequencing procedures were in full agreement with real-time PCR test results. Sequence analysis showed that the mutant gene had the expected base change compared to the wild type gene. The new procedure is not only very reliable but also faster than the techniques currently applied for diagnosis of the mutation.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2008.04.005