Identifiable universal fluorescent multiplex PCR equipped with capillary electrophoresis for genotyping of exons 1 to 5 in human red and green pigment genes.
Level 4 - case-series / case-control
Diagnostic method validation study evaluated against a reference standard in a small cross-sectional sample
PubMed 35033897 · doi:10.1016/j.talanta.2021.123199
What was done
Authors developed a forward specific primers with identifiable universal fluorescent multiplex PCR (FSIUFM-PCR) coupled with capillary electrophoresis to simultaneously amplify and label 11 gene targets (exons 1 to 5 in human red and green pigment genes, plus one internal standard). The assay was tested on genomic samples from 80 individuals (7 with color vision deficiencies and 73 with normal color vision) and compared directly against DNA sequencing.
What was found
Across all 80 individuals, the assay resolved the targeted exons. Among the 7 individuals with color vision deficiencies, 3 had only red exons 1 to 5, and 4 possessed specific red-green or green-red hybrid genes; 73 individuals had normal color vision profiles. All genotyping results demonstrated complete agreement with reference DNA sequencing. Specific diagnostic accuracy metrics (such as sensitivity, specificity, or confidence intervals) were not reported in the abstract.
Why it matters
The assay provides a single-reaction, high-resolution approach for detecting deletions and hybrid rearrangements in visual pigment genes, potentially speeding up molecular diagnosis of congenital red-green color blindness.
Limits
The clinical validation sample size was small (n = 80) and included only 7 affected individuals, preventing comprehensive assessment across diverse or rare hybrid alleles. The assay is limited to exons 1 through 5 (omitting exon 6), and quantitative analytical performance characteristics were not provided in the abstract.
Cited by
- supports Most red-green colorblind individuals are male due to the location of the gene mutation on the X chromosome.