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<b>A SENSITIVE AND HIGH-RESOLUTION METHOD FOR THE DETECTION OF MUTATIONS IN THE p53 GENE USING FLUORESCENCE-BASED </b><b>PCR-SSCP </b>

1995; Allied Academies; Volume: 16; Issue: 4 Linguagem: Inglês

10.2220/biomedres.16.273

1880-313X

Kiyonori Katsuragi, Wataru Chiba, Yoshito Matsubara, Sadao Ikeda, CHIE UETA, Moritoshi Kinoshita,

Molecular Biology Techniques and Applications

Mutations of the p53 gene are an important feature of neoplastic progression in humans.We have developed a simple and rapid method, fluorescence-based polymerase chain reaction single-strand conformation polymorphism (PCR-FSSCP), which uses an automated DNA sequencer and software, for detection of mutations in exons 5, 6, 7 and 8 of the p53 gene.Using this method, we analysed 63 specimens of cancerous tissue and normal tissue from lung cancer patients.Of the 63 patients of cancer, mutations were detected in 18 (28.5%).Moreover, in one of these 18 specimens, two mutated alleles were detected in exon 5.The frequencies of mutation were 14.3% in exon 5, 3.2% in exon 6 and 11.1% in exon 7.In contrast, mutations of the p53 gene were detected in 13 (20.6%) of the 63 specimens by PCR-SSCP using small polyacrylamide gels (8 cm x 9 cm) with ethidium bromide staining.