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• W2372738260 abstract "Objective To develop a real-time fluorescent quantitative PCR(FQ-PCR)combined with Sanger sequencing method for determination of epidermal growth factor receptor(EGFR),KRAS and BRAF gene mutations in colorectal cancer. Methods Primers and TaqMan probes were designed based on common mutations of EGFR,KRAS and BRAF gene in colorectal cancer for FQ-PCR amplification using the tumor tissue DNAs of 86 patients with colorectal cancer as templates. The PCR products were digested with SAP and EXOⅠ and purified by ethanol / sodium acetate method,then sequenced. The sequencing result was analyzed by Bioedit software. The mutations of EGFR, KRAS and BRAF genes were tested, and the results were compared with that of test for KRAS12 / 13 codon mutation by FQ-PCR. Results A method for determination of EGFR,KRAS and BRAF gene mutations in colorectal cancer by FQ-PCR combined with Sanger sequencing was successfully developed. The PCR amplification curves of various genes were in standardSshape,and the sequencing peaks were clear and stable. The probability of EGFR gene mutation in 86 colorectal cancer specimens was 2. 3%(2 / 86),both of which were found in exon 21,and expressed as L858 R and L861 Q respectively.However,the probability of KRAS gene mutation was 33. 7%(29 / 86),of which 79. 3%(23 / 29)in codon 12(G12Dand G12V),10. 3%(3 / 29)in codon 13(G13D),and 3. 4% in both codons 12 and 13. No mutations in codon 61 were observed,while those in codon 146 were observed in 2 of 29 specimens(6. 9%),both of which were expressed as A146 V. No mutations of BRAF gene were found. The probability of KRAS12 / 13 mutation determined by FQ-PCR was insignificantly higher than that by FQ-PCR combined with Sanger sequencing(P 0. 05),with a total coincidence rate of 97. 7%(84 / 86). Conclusion Real-time FQ-PCR combined with Sanger sequencing may be used for rapid determination of EGFR,KRAS and BRAF gene mutations in colorectal cancer,which was stable and reliable." @default.
• W2372738260 created "2016-06-24" @default.
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• W2372738260 date "2014-01-01" @default.
• W2372738260 modified "2023-09-23" @default.
• W2372738260 title "Test for epidermal growth factor receptor,KRAS and BRAF gene mutations in colorectal cancer by FQ-PCR combined with Sanger sequencing" @default.
• W2372738260 hasPublicationYear "2014" @default.
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