Yuefan Xu | PubMed Health

PubMed2002

[Restriction endonucleases fingerprinting-single strand conformation polymorphism: an efficient method to screen mutations in long segments].

OBJECTIVE: To develop a simple, cheap and efficient restriction endonucleases fingerprinting-single strand conformation polymorphism(REF-SSCP) method applied to screen for mutations in long segments. METHODS: The 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. RESULTS: No 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%. CONCLUSION: The present REF-SSCP method is applicable to screen mutations in long segment DNA of mass specimens.

Connexin 26↗

PubMed2002

[Mutation analysis of Cx26 gene in Chinese hereditary nonsyndromic deafness sufferers].

OBJECTIVE: To analyze the mutations in the code region of Cx26 gene in Chinese hereditary nonsyndromic hearing impairment (NSHI) sufferers. METHODS: Thirty-three cases (29 cases in the families of 8 students who were picked out from the Deafness and Muteness School of Tianjin, 2 cases as control and 2 normal cases of genetic counseling) were included in this study. The blood samples were obtained to distill the DNA templates. Using polymerase chain reaction (PCR), the code region of Cx26 gene was amplified. The mutations were screened by restriction endonucleases fingerprinting-single strand conformation polymorphism (REF-SSCP). Afterwards we inspected the polymorphous changes or mutations of these segments with DNA sequence. RESULTS: There were 30 cases with the nucleotide changes in the Cx26 code region. The rate was 90.9% (30/33). Eight kinds of mutations were found, 79G-->A, 109G-->A, 161A-->T, 235delC, 240G-->A, 341A-->G, 571T-->C and 608T-->C. 161A-->T, 240G-->A and 571T-->C were detected primarily. There were 3 cases with 235delC in 22 deafness sufferers and the rate was 13.64% (3/22). CONCLUSIONS: The 235delC of Cx26 gene is the main mutation in Chinese NSHI sufferers, and in NSHI cases many polymorphous changes exist.

Adolescent↗