Molecular detection of autosomal-dominant feline polycystic kidney disease by multiplex amplification refractory mutation system polymerase chain reaction.
Feline autosomal-dominant polycystic kidney disease (ADPKD), with its characteristic growth of fluid-filled cysts of different sizes, is the most prevalent inherited genetic disease of cats. The point mutation (C-->A transversion) in exon 29 of the PKD1 gene is known to contribute to ADPKD development and can thus serve as a target for the molecular genetic diagnosis of ADPKD. To this end, a simple amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) was designed with 3 primers: 2 forward primers specifically targeting either the mutant or normal allele, and 1 universal reverse primer for amplification of both alleles. The new method was tested on the DNA from 35 feline blood samples, which included 15 mutant cats and 20 wild type cats. As verified by direct DNA sequencing, both sensitivity and specificity of this tri-primer ARMS PCR were 100%. As the multiplex ARMS PCR test can be performed in a single PCR reaction without other post-PCR procedures, it is a simple and accurate method for molecular studies of feline ADPKD.
Lee
Ya-Jane
YJ
Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung 402, Taiwan.
Chen
Hsin-Yu
HY
Wong
Min-Liang
ML
Hsu
Wei-Li
WL
D000818:Animals; D002371:Cat Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics* / Q000473:pathology; D002415:Cats; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D005091:Exons / Q000235:genetics; D005784:Gene Amplification; D015994:Incidence; D009154:Mutation; D016891:Polycystic Kidney, Autosomal Dominant / Q000453:epidemiology / Q000235:genetics* / Q000473:pathology / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods; D013045:Species Specificity; D050396:TRPP Cation Channels / Q000235:genetics
D004247:DNA; D017931:DNA Primers; D050396:TRPP Cation Channels