Applications of gene-chip for the detection of mutations in cTnI gene associated with FHCM

摘要

Mutations in cardiac troponin I (cTnl) gene were assessed based on gene-chip technology. Special probes were designed to fabricate the low-density gene-chip, which could detect the mutations in exons 3,5,7, and 8 of the cTnl gene simultaneously. For each exon, two oligonucleotide sequences labeled with fluorescein at the 5'-end were designed, one (oligonucleotide I) simulating the wild type and the other (oligonucleotide II) simulating the mutant. Oligonucleotides I and I were mixed together to simulate the heterozygote. After optimizing the hybridization protocols, the fabricated gene-chip could detect the mutations in the exons of the cTnI gene with relative high sensitivity and specificity. The fully complementary probe gave a fluorescent signal almost 50 percent stronger than that of the one-base mismatched one, which is in accordance with the result from a theoretical estimate. An applicable special gene-chip is available to investigate and diagnose familial hypertrophic cardiomyopathy (FHCM) after further improvement.