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Title: KRAS and BRAF mutation analysis can be reliably performed on aspirated cytological specimens of metastatic colorectal carcinoma
Authors: Pang, N.K.B.
Nga, M.E.
Chin, S.Y. 
Ismail, T.M.
Lim, G.L.
Soong, R. 
Salto-Tellez, M. 
Keywords: Anti-EGFR monoclonal antibody therapy
Colorectal adenocarcinoma
Cytological specimens
Direct sequencing
DNA extraction
KRAS and BRAF mutation status
Issue Date: Dec-2011
Citation: Pang, N.K.B., Nga, M.E., Chin, S.Y., Ismail, T.M., Lim, G.L., Soong, R., Salto-Tellez, M. (2011-12). KRAS and BRAF mutation analysis can be reliably performed on aspirated cytological specimens of metastatic colorectal carcinoma. Cytopathology 22 (6) : 358-364. ScholarBank@NUS Repository.
Abstract: Background: Sanger sequencing is one of several reliable methods in use to detect KRAS and BRAF mutations to facilitate clinical patient selection for anti-epidermal growth factor receptor (EGFR) monoclonal antibody therapy in unresectable metastatic colorectal adenocarcinoma (CRC). Most analyses are made on pretreatment biopsy or resection specimens. There is a scarcity of published studies on the suitability of cytological samples for KRAS testing in this setting. Methods: DNA extraction was attempted on 11 search-retrieved paired cases of histological resections or excisions of CRC and their corresponding cytological samples (representing metastases) and tested for KRAS mutations in exon 2 and 3, as well as BRAF exon 15 mutations by Sanger sequencing. Only KRAS wild-type cases were subjected to BRAF analysis because this is the setting with true diagnostic value, as these mutations are mutually exclusive. Results: Of the 11 paired cases analysed, only eight histology cases showed satisfactory DNA quality for sequencing. Thus, only eight of the corresponding cytology cases were analysed. Seven of the eight cases tested showed the same KRAS genotype on both the aspirated cytology specimen of metastatic carcinoma and the primary tumour (histological specimen), from which we derive an overall concordance rate of 87.5%. The single discordant case was likely to be a true difference as it was demonstrated again on repeat testing of both samples. No BRAF mutations were detected on the four KRAS wild-type cases. Conclusion: A range of cytological samples are suitable for KRAS and BRAF mutation testing, be it from previously stained preparations or cell blocks. These samples would be highly valuable in cases where cytological samples are the only material available for mutation testing. © 2010 Blackwell Publishing Ltd.
Source Title: Cytopathology
ISSN: 09565507
DOI: 10.1111/j.1365-2303.2010.00812.x
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