Non isotopic PCR cold SSCP of p53 gene mutations in gliomas using the Dcode system
M.N. Isa1*, A.A. Yusoff1, M.R. Sidek1 and J.M. Abdullah2
1Human Genome Centre, Health Campus, 2Neuroscience Unit, School of Medical Sciences, Health Campus,
Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
Received 8 June 2001 / Accepted 16 November 2001
Abstract.
The p53 tumor suppressor gene is the most commonly mutated gene identified in human cancers. The abnormal changes of this gene could be detected by several methods. In the present study, we have developed a procedure for non isotopic PCR cold SSCP using the Dcode Universal Mutation Detection System and applied it to the detection of p53 gene mutations in 33 Malaysian patients with surgically removed gliomas tissues. Our research showed that the optimum buffer temperature for 'cold' SSCP analysis was 10oC and highly specific for the PCR products between 100-250 bp. Mutations of the p53 gene were detected in 11 cases (33%) of 33 surgical specimens of gliomas. Additionally, p53 mutations occurred most frequently at exon 7 (45.5%), followed by exon 8 (36.4%) and exon 6 (18. 1 %). Our results indicate that the utility of the non isotopic PCR cold SSCP is a convenient, rapid and sensitive screening assay for detecting p53 gene mutations
Keywords: p53 gene, gliomas, Dcode, cold SSCP
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