Universität zu Köln

Mairinger, Fabian D., Vollbrecht, Claudia ORCID: 0000-0002-0861-001X, Streubel, Anna, Roth, Andreas, Landt, Olfert, Walter, Henry F. R., Kollmeier, Jens ORCID: 0000-0002-8048-3895 and Mairinger, Thomas (2014). The COLD-PCR Approach for Early and Cost-Effective Detection of Tyrosine Kinase Inhibitor Resistance Mutations in EGFR-Positive Non-Small Cell Lung Cancer. Appl. Immunohistochem., 22 (2). S. 114 - 119. PHILADELPHIA: LIPPINCOTT WILLIAMS & WILKINS. ISSN 1533-4058

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Abstract

Background:Activating epidermal growth factor receptor (EGFR) gene mutations can be successfully treated by EGFR tyrosine kinase inhibitors (EGFR-TKIs), but nearly 50% of all patients' exhibit progression of the disease until treatment because of T790M mutations. It is proposed that this is mostly caused by therapy-resistant tumor clones harboring a T790M mutation. Until now no cost-effective routine-diagnostic method for EGFR-resistance mutation status analysis is available leaving long-time response to TKI treatment to chance. Unambiguous identification of T790M EGFR mutations is mandatory to optimize initial treatment strategies.Materials and Methods:Artificial EGFR T790M mutations and human wild-type gDNA were prepared in several dilution series. Preferential amplification using coamplification at lower denaturation temperature-PCR (COLD-PCR) of the mutant sequence and subsequent HybProbe melting curve detection or pyrosequencing were performed in comparison to normal processing.Results:COLD-PCR-based amplification allowed the detection of 0.125% T790M mutant DNA in a background of wild-type DNA in comparison to 5% while normal processing. These results were reproducible.Conclusions:COLD-PCR is a powerful and cost-effective tool for routine diagnostic to detect underrepresented tumor clones in clinical samples. A diagnostic tool for unambiguous identification of T790M-mutated minor tumor clones is now available enabling optimized therapy.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Mairinger, Fabian D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Vollbrecht, Claudia UNSPECIFIED orcid.org/0000-0002-0861-001X UNSPECIFIED Streubel, Anna UNSPECIFIED UNSPECIFIED UNSPECIFIED Roth, Andreas UNSPECIFIED UNSPECIFIED UNSPECIFIED Landt, Olfert UNSPECIFIED UNSPECIFIED UNSPECIFIED Walter, Henry F. R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kollmeier, Jens UNSPECIFIED orcid.org/0000-0002-8048-3895 UNSPECIFIED Mairinger, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-446148
DOI:	10.1097/PDM.0b013e31829a638d
Journal or Publication Title:	Appl. Immunohistochem.
Volume:	22
Number:	2
Page Range:	S. 114 - 119
Date:	2014
Publisher:	LIPPINCOTT WILLIAMS & WILKINS
Place of Publication:	PHILADELPHIA
ISSN:	1533-4058
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language GROWTH-FACTOR-RECEPTOR; KRAS MUTATIONS; GENE; AMPLIFICATION Multiple languages Anatomy & Morphology; Medical Laboratory Technology; Pathology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/44614