Medline Abstract Wild-type blocking polymerase chain reaction for detection of single nucleotide minority mutations from clinical specimens. PL Dominguez and MS Kolodney Oncogene, October 13, 2005; 24(45): 6830-4. Abstract Full text via Infotrieve Alert me when cited Find more like this   Department of Medicine, Division of Dermatology, Harbor - UCLA Medical Center, 1000 W Carson St, Box 459, Torrance, CA 90509, USA.		Download to Citation Manager Alert me when this article is cited PubMed Citation Related Articles in PubMed Order Full text via Infotrieve This article has been cited by other articles
Detection and sequencing of mutations from clinical specimens is often complicated by the presence of an excess of nonmutated cells. To facilitate the detection and sequencing of minority mutations from clinical specimens, we developed wild-type blocking polymerase chain reaction (WTB-PCR). This technique allows sensitive detection of minority mutations in a tissue sample containing excess wild-type DNA. In WTB-PCR, a nonextendable locked nucleic acid (LNA) oligonucleotide binds tightly to a region of wild-type DNA known to develop point mutations. This LNA sequence blocks amplification of wild-type DNA during PCR while permitting amplification of mutant exon 15. Our results show that the LNA blocking oligonucleotide inhibits amplification of wild-type DNA in a dose-dependent manner. WTB-PCR was able to detect mutant DNA in clinical samples of melanoma tissue containing an excess of nonmelanoma cells. This method was also able to detect small amounts of point mutated or tandem mutated DNA diluted with a much larger concentration of wild-type DNA. This rapid and simple assay overcomes the limitations of current methods to detect minority mutations. The potential applications of WTB-PCR include early diagnosis and prognosis of various cancers. Erratum in Oncogene. 2006 Jan 26;25(4):656 Publication Types: Journal article Research support, non-u.s. gov't MeSH Terms: Base Sequence DNA Primers Mutation* Nucleotides Polymerase Chain Reaction PMID: 16116485

This article has been cited by other articles:

			REGULAR ARTICLES: Rapid Method for Detection of Mutations in the Nucleophosmin Gene in Acute Myeloid Leukemia Todd S. Laughlin, Michael W. Becker, Jane L. Liesveld, Deborah A. Mulford, Camille N. Abboud, Patrick Brown, and Paul G. Rothberg J. Mol. Diagn., July 1, 2008; 10: 338 - 345.				Abstract Full text PDF Find more like this

			EDITORIALS: Epidermal Growth Factor Receptor Mutation Testing in Lung Cancer: Searching for the Ideal Method William Pao and Marc Ladanyi Clin. Cancer Res., September 1, 2007; 13: 4954 - 4955.				Full text PDF Find more like this Related Topics

			CANCER DIAGNOSTICS: Sensitive Detection of KIT D816V in Patients with Mastocytosis Angela Tan, David Westerman, Grant A. McArthur, Kevin Lynch, Paul Waring, and Alexander Dobrovic Clin. Chem., December 1, 2006; 52: 2250 - 2257.				Abstract Full text PDF extra: Data Supplement Find more like this

			REGULAR ARTICLES: Allele-Specific Polymerase Chain Reaction for the Imatinib-Resistant KIT D816V and D816F Mutations in Mastocytosis and Acute Myelogenous Leukemia Christopher L. Corless, Patina Harrell, Mario Lacouture, Troy Bainbridge, Claudia Le, Ken Gatter, Clifton White, Jr, Scott Granter, and Michael C. Heinrich J. Mol. Diagn., November 1, 2006; 8: 604 - 612.				Abstract Full text PDF Find more like this Related Topics

			LETTERS TO THE EDITOR: Combined Locked Nucleic Acid and Molecular Beacon Technologies for Sensitive Detection of the JAK2V617F Somatic Single-Base Sequence Variant Pierre Sidon, Pierre Heimann, Frédéric Lambert, Barbara Dessars, Valérie Robin, and Hakim El Housni Clin. Chem., July 1, 2006; 52: 1436 - 1438.				Extract Full text PDF extra: Supplemental Data extra: 066886.Data Supplements Find more like this Related Topics