HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (21) Citing Articles via Google Scholar Google Scholar Articles by Lopez-Crapez, E. Articles by Grenier, J. Search for Related Content PubMed PubMed Citation Articles by Lopez-Crapez, E. Articles by Grenier, J. Related Collections Molecular Diagnostics and Genetics Automation and Analytical Techniques

K-ras Mutation Detection by Hybridization to a Polypyrrole DNA Chip

¹ CRLC Val d’Aurelle Paul-Lamarque, Centre de Recherche en Cancérologie, Parc Euromédecine, 34298 Montpellier Cedex 5, France.

² CIS bio international, 91192 Gif sur Yvette Cedex, France.
^a Author for correspondence. Fax 33-4-67-63-28-73; e-mail ecrapez{at}valdorel.fnclcc.fr.

Background: Detection of mutations in cancer-related genes is of major importance for both basic knowledge and clinical practice. Several strategies have been developed to diagnose these alterations. We describe a method based on polypyrrole DNA chip technology to detect K-ras gene mutations in tumors.

Methods: An oligodeoxynucleotide array was constructed on a silicon device by copolymerization of 5'-pyrrole-labeled oligodeoxynucleotides and pyrrole. The samples to be analyzed were then amplified by PCR, and the single-stranded biotin-labeled amplified DNA was specifically hybridized to the addressed probes. Perfectly matched duplexes were detected by fluorescence microscopy using R-phycoerythrin as the detection label. The developed methodology was applied to genotype assignment of K-ras in human samples. The genotypes of 75 DNA genomic samples from colorectal cancer patients were analyzed side by side using direct DNA sequencing and a polypyrrole DNA chip.

Results: The chip method unequivocally defined all of the genotypes. Mutations present at <10% of the wild-type DNA concentration could be distinguished.

Conclusions: This probe array assay is a rapid and reliable procedure that may be used to detect mutations.

The following articles in journals at HighWire Press have cited this article:

				A. Halperin, A. Buhot, and E. B. Zhulina Hybridization Isotherms of DNA Microarrays and the Quantification of Mutation Studies Clin. Chem., December 1, 2004; 50(12): 2254 - 2262. [Abstract] [Full Text] [PDF]

				J.-H. Park, I.-J. Kim, H. C. Kang, Y. Shin, H.-W. Park, S.-G. Jang, J.-L. Ku, S.-B. Lim, S.-Y. Jeong, and J.-G. Park Oligonucleotide Microarray-Based Mutation Detection of the K-ras Gene in Colorectal Cancers with Use of Competitive DNA Hybridization Clin. Chem., September 1, 2004; 50(9): 1688 - 1691. [Full Text] [PDF]

				L. Prix, P. Uciechowski, B. Bockmann, M. Giesing, and A. J. Schuetz Diagnostic Biochip Array for Fast and Sensitive Detection of K-ras Mutations in Stool Clin. Chem., March 1, 2002; 48(3): 428 - 435. [Abstract] [Full Text] [PDF]

				I.-J. Kim, H. C. Kang, J.-H. Park, J.-L. Ku, J.-S. Lee, H.-J. Kwon, K.-A. Yoon, S. C. Heo, H.-Y. Yang, B. Y. Cho, et al. RET Oligonucleotide Microarray for the Detection of RET Mutations in Multiple Endocrine Neoplasia Type 2 Syndromes Clin. Cancer Res., February 1, 2002; 8(2): 457 - 463. [Abstract] [Full Text] [PDF]