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Title: | Development of modified mismatch PCR-RFLP to screen mutations in codon 12 and 13 of K- ras gene of colorectal (CRC) patients in Sri Lanka |
Authors: | Dhilhani, M.F.F. de Zoysa, M.I.M. Chandrasekharan, N.V. Gunawardene, Y.I.N.S. Lokuhetti, M.D.S. Dassanayake, R.S. |
Keywords: | colorectal (CRC) |
Issue Date: | 2015 |
Publisher: | Sri lanka Medical Association |
Citation: | Proceedings of the Sri Lanka Medical Association, Anniversary Academic Sessions. 2015; 60(sup 1): 185 |
Abstract: | INTRODUCTION AND OBJECTIVES: Mutations in K-ras codon 12, 13 of exon 2 are known to affect prognosis and impart resistance to anti EGFR monoclonal antibody therapy in CRC. Although several diagnostic tools have been developed for K-ras mutation testing, these procedures are too expensive or time consuming. Oufaim was to develop an effective, reliable and inexpensive method for the detection of K-ras mutations in codons 12 and 13 of exon 2 in CRC patients in Sri Lanka, and to relate the mutational status to liver metastasis, METHOD: The mismatch PCR-RFLP was developed and used to screen mutations in codon 12 and 13 for DMA isolated from paraffinized tumour tissue of 30 CRC patients followed up for 5 year after surgery to detect liver metastasis. Cross-tabulations were generated between K-ras mutations and the metastatic status. The Chi Square test was used to indicate statistical significance of the association. RESULTS: Analysis of banding pattern obtained from restriction digestion of PCR amplified region containing codon 12 and/or 13 of KRAS gene of 14(46.6%) CRC patients revealed the presence of mutations. Of the 30 patients, 13(43.3%) had developed liver metastases. There was a significant association between the presence of a K-ros mutation and the occurrence of liver metastasis (X2=4.693, p=0.003). CONCLUSION: This mismatch PCR-RFLP protocol is a suitable method to screen codon 12 and 13 mutation of K-ros gene to predict liver metastasis. Presence of these mutations is associated with the occurrence of liver metastasis during the first 5 years after surgery. |
Description: | Poster Presentation Abstract (PP50), 128th Annual Scientific Sessions, Sri Lanka Medical Association, 6th-8th July 2015 Colombo, Sri Lanka |
URI: | http://repository.kln.ac.lk/handle/123456789/10642 |
Appears in Collections: | Conference Papers |
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