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《Chinese Journal of Clinical Medicine》 2006-01
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Validation of Two New Methods for Mutation Detection in Sporadic Ovarian Cancer

DU Ming FENG Youji Hospital of Obstetrics & Gynecology, Fudan University, Shanghai 200011  
Objective: Two lately developed methods for mutation detection, DHPLC (denaturing high-performance liquid chromatography) and F-DDF (fluorescent dideoxy fingerprinting) have been tested for their suitability by analysing heterogeneous tumor material. Methods:For comparison of both techniques 50 sporadic ovarian cancers (OC) including 18 known p53 mutations (blind study) were examined analysing exons 5-8 by DHPLC and by F-DDF (total 200 PCR-fragments). The conventional DDF was adapted to an ALF express?DNA sequencing system using Cy5-labeled primers for single-strand sequencing reaction with ddGTP or ddTTP. For DHPLC analyses the WAVE system (Transgenomic) was used. Results:All 18 mutations could be detected by DHPLC analysis (sensitivity: 100%) without any false positive/negative results whereas 4 mutations were missed by F-DDF (sensitivity: 78%). Interpretation of F-DDF results is more difficult therefore a higher number of false positive results were obtained. Conclusion:DHPLC is a reliable high throughput technique for mutation detection in heterogeneous tumor material. Mutation detection by F-DDF revealed only a sensitivity of 78% and a lower specificity. However in contrast to DHPLC analysis no additional equipment is necessary for F-DDF analysis.
【Fund】: 博士后科学基金资助(2003033278)
【CateGory Index】: R737.31
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