An improved diagnostic test for rod cone dysplasia 1 (rcd1) using allele-specific polymerase chain reaction.

Abstract:

PURPOSE:To develop an improved diagnostic test for rod-cone dysplasia type 1 (rcd1). The rcd1 phenotype is an early onset, autosomal recessive disease caused by a mutation in the canine rod cyclic GMP phosphodiesterase beta-subunit (PDE6B) gene. A G to A transition in codon 807 at nucleotide position 2420 results in a stop codon. This is the only disease causing mutation detected so far in the canine PDE6B gene. METHODS:Allele specific primers were designed in which the 3 end had the nucleotide corresponding to either the wild type or the mutant rcd1 allele. PCR was done using the allele specific primers in combination with a common primer complementary to the opposite strand to distinguish between the wild type and the rcd1 alleles. RESULTS:The wild type and rcd1 alleles were identified successfully in two independent ASPCRs done with two different sets of allele specific primers. Further, both alleles could be amplified in a single tube and distinguished based on the size difference of the PCR products using one allele specific primer of altered length by the addition of a 9 nucleotide long linker. CONCLUSIONS:We have developed an improved diagnostic test for the disease based on ASPCR such that the presence or absence of different size amplified fragments provides direct determination of the genotype. In contrast to previously reported diagnostic tests, this method is more efficient because it eliminates the need for any further manipulation of the PCR product.

journal_name

Curr Eye Res

journal_title

Current eye research

authors

Ray K,Tejero MD,Baldwin VJ,Aguirre GD

doi

10.3109/02713689609000770

subject

Has Abstract

pub_date

1996-05-01 00:00:00

pages

583-7

issue

5

eissn

0271-3683

issn

1460-2202

journal_volume

15

pub_type

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