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Journal of Clinical & Medical Genomics
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Short Communication

Targeting Cancer Related Genes by Multiplex PCR Followed by High Throughput Parallel Sequencing

Vollbrecht Claudia1*, Mairinger Fabian2, Schweighofer Carmen Diana3, Heukamp Lukas1, Merkelbach-Bruse Sabine1,Büttner Reinhard1 and Margarete Odenthal1*

1 Institute of Pathology, University Hospital Cologne, Germany

2 Institute of Pathology, University Hospital Essen, University of Duisburg-Essen, Germany

3 Schweighofer, Carmen Diana; Medical Clinic I, University Hospital Cologne, Germany

*Corresponding Author:
Margarete Odenthal
Institute of Pathology
University Hospital Cologne Kerpener Str. 62
50937 Cologne, Germany
Tel: +49(0)2214786367
E-mail: [email protected]
 
Claudia Vollbrecht
Institute of Pathology
University Hospital Cologne
Kerpener Str. 62, 50937 Cologne, Germany
Tel: +49(0)22147888936
E-mail: [email protected]

Received Date: December 10, 2013; Accepted Date: February 21, 2014; Published Date: March 01, 2014

Citation: Claudia V, Fabian M, Diana SC, Lukas H, Sabine MB, et al. (2014) Targeting Cancer Related Genes by Multiplex PCR Followed by High Throughput Parallel Sequencing. Int J Genomic Med 2:115. doi: 10.4172/2332-0672.1000115

Copyright: © 2014 Claudia V, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

The detection of a wide range of genomic alterations plays an important role in the diagnostics improving individual therapeutic approaches of cancer patients. Technologies that help to identify therapeutic relevant targets in tumor samples are a major factor on the way to a personalized medicine. The number of predictive and prognostic markers that influence the therapeutic outcome is continuously increasing. Therefore, parallel sequencing also named next generation sequencing (NGS), allowing the simultaneous analysis of numerous cancer related hotspots in many patients starting from a limited amount of DNA, is urgently needed to be established in cancer diagnostics. Different methods of target library preparation are commercially available and offer the opportunity to sequence tumor relevant hotspot mutations in a panel of genes.

In the present study, a multiplex PCR approach, targeting a lung cancer and a leukemia gene panel, each consisting of 20 disease and therapy relevant genes, was investigated. Twelve formalin fixed and paraffin embedded lung tumors and twelve native Chronic Lymphocytic Leukemia (CLL) samples, respectively, were analyzed. Samples were sequenced on the MiSeq sequencer platform.

The results showed a very high quality of each run. In spite of the low DNA input, each multiplex approach allowed a simultaneous analysis of 20 genes, in total covered by around 1,000 amplicons, in up to twelve cancer samples.

Thus, the application of NGS on amplicon targets revealed an excellent performance in detecting a wide range of genetic alterations, combined with a high sensitivity.

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