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The p53 tumor suppressor gene is a 16-20-kb section of cellular DNA located on the short arm of human chromosome 17 at position 17 P 13.1. Allelic deletions and/or point mutations in p53 gene are now known to be associated with the development of carcinogenesis. A hallmark of p53 is that both alleles are generally altered during transformation, which usually represents a loss of heterozygosity (LOH). In this study 30 normal dental students and 22 oral cancer patients were collected from the affiliated hospital of Chung Shan Medical and Dental College, Taichung, Taiwan. Extractions of DNA from the buccal mucosa or cancer surface were sampled by cytology brush. The two polymorphic restriction sites exon 4 and intron 6 within the p53 gene were amplified with polymerase chain reactions followed by restriction fragment length polymorphism assay. In heterozygous individuals, 66% of oral cancers demonstrated loss of p53 gene heterozygosity at the exon 4 site, and 50% showed LOH at the intron 6 site. These results indicate that inactivation of p53 gene is associated with development and/or progression of oral cancer. The essential advantages of oral exfoliative cytology are the non-invasiveness, painlessness, rapidity, ease and cost-effectiveness of cell sampling and DNA extraction. Furthermore, this experimental assay might be useful for preliminary screening of carcinogenesis in human beings.