OBJECTIVETo develop a simple, cheap and efficient restriction endonucleases fingerprinting-single strand conformation polymorphism(REF-SSCP) method applied to screen for mutations in long segments.

METHODSThe genomic DNA of Cx26 gene segment of the patients with deafness was amplified. The amplification products were screened with SSCP and REF-SSCP technique and DNA sequencing to evaluate and compare the effect on detection of mutations in long segments.

RESULTSNo different band was found in 724 bp segment in SSCP examination. Three kinds of different bands were discovered in REF-SSCP examination and the 79 G -->A mutation detected by DNA sequencing were accorded with the REF-SSCP bands entirely. The rate of detection was 100%.

CONCLUSIONThe present REF-SSCP method is applicable to screen mutations in long segment DNA of mass specimens.

Connexin 26 ; Connexins ; genetics ; DNA Fingerprinting ; methods ; DNA Restriction Enzymes ; Deafness ; genetics ; Genetic Testing ; methods ; Humans ; Mutation ; Polymorphism, Single-Stranded Conformational ; Sensitivity and Specificity