In:
Cancer Research, American Association for Cancer Research (AACR), Vol. 73, No. 8_Supplement ( 2013-04-15), p. 4223-4223
Abstract:
We previously described the utility of the CEE-Selector assay for the sensitive detection of the EGFR T790M mutation in plasma samples from lung cancer patients. The assay combined the CEE-Selector assay with melt curve analysis of the mutant PCR product, followed by Sanger sequencing to verify the presence of the mutation. We showed that the assay selectively amplified the mutant sequence compared to wild type by & gt;100,000 fold. We have now expanded the CEE-Selector assay to include additional mutations relevant to patient care (KRAS, BRAF V600E, EGFR L858R). We are presenting data of allele/mutation-specific as well as “footprint” versions of the CEE-Selector assay. The “footprint” version of the assay provides for the interrogation of “hot spot” regions of the genome where multiple cancer relevant mutations are located within a small region, as in the case of KRAS codons 12 and 13 where the CEE-Selector assay is sensitive to all known mutations. Combining the “footprint” version of the CEE-Selector assay with next-generation sequencing, allows us to simultaneously detect, with extremely high sensitivity, a broad array of mutations of interest. Additionally, when used with barcoding, samples from multiple patients can be interrogated simultaneously. The combination of the CEE-Selector assay with next-generation sequencing increases the sensitivity of sequencing by a factor of & gt;1000 and allows for simultaneous sensitive detection of multiple mutations from complex samples, like plasma, where the mutations may be very rare. With the increases in sensitivity made possible using the CEE-Selector assay, the limiting factor for the sensitive detection of mutations/alleles is determined fundamentally by the ability to obtain at least a single copy of the mutant allele within the sample under interrogation. Citation Format: Vassilios Alexiadis, Tim Watanaskul, Lyle Arnold. Combining the CEE-selector assay with sequencing for sensitive mutation detection in cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4223. doi:10.1158/1538-7445.AM2013-4223
Type of Medium:
Online Resource
ISSN:
0008-5472
,
1538-7445
DOI:
10.1158/1538-7445.AM2013-4223
Language:
English
Publisher:
American Association for Cancer Research (AACR)
Publication Date:
2013
detail.hit.zdb_id:
2036785-5
detail.hit.zdb_id:
1432-1
detail.hit.zdb_id:
410466-3
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