Universität zu Köln

Mairinger, Fabian D., Walter, Robert F. H., Vollbrecht, Claudia ORCID: 0000-0002-0861-001X, Hager, Thomas, Worm, Karl, Ting, Saskia ORCID: 0000-0003-1415-7714, Wohlschlaeger, Jeremias, Zarogoulidis, Paul, Zarogoulidis, Konstantinos and Schmid, Kurt W. (2014). Isothermal multiple displacement amplification: a methodical approach enhancing molecular routine diagnostics of microcarcinomas and small biopsies. OncoTargets Ther., 7. S. 1441 - 1448. ALBANY: DOVE MEDICAL PRESS LTD. ISSN 1178-6930

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Abstract

Background and methods: Isothermal multiple displacement amplification (IMDA) can be a powerful tool in molecular routine diagnostics for homogeneous and sequence-independent whole-genome amplification of notably small tumor samples, eg, microcarcinomas and biopsies containing a small amount of tumor. Currently, this method is not well established in pathology laboratories. We designed a study to confirm the feasibility and convenience of this method for routine diagnostics with formalin-fixed, paraffin-embedded samples prepared by laser-capture microdissection. Results: A total of 250 mu g DNA (concentration 5 mu g/mu L) was generated by amplification over a period of 8 hours with a material input of approximately 25 cells, approximately equivalent to 175 pg of genomic DNA. In the generated DNA, a representation of all chromosomes could be shown and the presence of elected genes relevant for diagnosis in clinical samples could be proven. Mutational analysis of clinical samples could be performed without any difficulty and showed concordance with earlier diagnostic findings. Conclusion: We established the feasibility and convenience of IMDA for routine diagnostics. We also showed that small amounts of DNA, which were not analyzable with current molecular methods, could be sufficient for a wide field of applications in molecular routine diagnostics when they are preamplified with IMDA.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Mairinger, Fabian D. UNSPECIFIED UNSPECIFIED UNSPECIFIED Walter, Robert F. H. UNSPECIFIED UNSPECIFIED UNSPECIFIED Vollbrecht, Claudia UNSPECIFIED orcid.org/0000-0002-0861-001X UNSPECIFIED Hager, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED Worm, Karl UNSPECIFIED UNSPECIFIED UNSPECIFIED Ting, Saskia UNSPECIFIED orcid.org/0000-0003-1415-7714 UNSPECIFIED Wohlschlaeger, Jeremias UNSPECIFIED UNSPECIFIED UNSPECIFIED Zarogoulidis, Paul UNSPECIFIED UNSPECIFIED UNSPECIFIED Zarogoulidis, Konstantinos UNSPECIFIED UNSPECIFIED UNSPECIFIED Schmid, Kurt W. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-449850
DOI:	10.2147/OTT.S65144
Journal or Publication Title:	OncoTargets Ther.
Volume:	7
Page Range:	S. 1441 - 1448
Date:	2014
Publisher:	DOVE MEDICAL PRESS LTD
Place of Publication:	ALBANY
ISSN:	1178-6930
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language WHOLE GENOME AMPLIFICATION; SINGLE-NUCLEOTIDE POLYMORPHISM; DNA AMPLIFICATION; DEGRADED DNA; POLYMERASE; HYBRIDIZATION; CRYOBIOPSY; INCREASES; TISSUES; SAMPLES Multiple languages Biotechnology & Applied Microbiology; Oncology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/44985