Journal article
Detection of somatic mutations by high-resolution DNA melting (HRM) analysis in multiple cancers
PloS one, Vol.6(1), pp.e14522-e14522
01/17/2011
DOI: 10.1371/journal.pone.0014522
PMCID: PMC3022009
PMID: 21264207
Abstract
Identification of somatic mutations in cancer is a major goal for understanding and monitoring the events related to cancer initiation and progression. High resolution melting (HRM) curve analysis represents a fast, post-PCR high-throughput method for scanning somatic sequence alterations in target genes. The aim of this study was to assess the sensitivity and specificity of HRM analysis for tumor mutation screening in a range of tumor samples, which included 216 frozen pediatric small rounded blue-cell tumors as well as 180 paraffin-embedded tumors from breast, endometrial and ovarian cancers (60 of each). HRM analysis was performed in exons of the following candidate genes known to harbor established commonly observed mutations: PIK3CA, ERBB2, KRAS, TP53, EGFR, BRAF, GATA3, and FGFR3. Bi-directional sequencing analysis was used to determine the accuracy of the HRM analysis. For the 39 mutations observed in frozen samples, the sensitivity and specificity of HRM analysis were 97% and 87%, respectively. There were 67 mutation/variants in the paraffin-embedded samples, and the sensitivity and specificity for the HRM analysis were 88% and 80%, respectively. Paraffin-embedded samples require higher quantity of purified DNA for high performance. In summary, HRM analysis is a promising moderate-throughput screening test for mutations among known candidate genomic regions. Although the overall accuracy appears to be better in frozen specimens, somatic alterations were detected in DNA extracted from paraffin-embedded samples.
Details
- Title: Subtitle
- Detection of somatic mutations by high-resolution DNA melting (HRM) analysis in multiple cancers
- Creators
- Jesus Gonzalez-Bosquet - Laboratory of Translational Genomics, Department of Health and Human Services, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of AmericaJacob CalceiJun S WeiMontserrat Garcia-ClosasMark E ShermanStephen HewittJoseph VockleyJolanta LissowskaHannah P YangJaved KhanStephen Chanock
- Resource Type
- Journal article
- Publication Details
- PloS one, Vol.6(1), pp.e14522-e14522
- DOI
- 10.1371/journal.pone.0014522
- PMID
- 21264207
- PMCID
- PMC3022009
- NLM abbreviation
- PLoS One
- ISSN
- 1932-6203
- eISSN
- 1932-6203
- Publisher
- United States
- Language
- English
- Date published
- 01/17/2011
- Academic Unit
- Obstetrics and Gynecology
- Record Identifier
- 9983930308102771
Metrics
17 Record Views