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Seifarth, W.* ; Frank, O.* ; Schreml, J.* ; Leib-Mösch, C.

RetroArray - a comprehensive diagnostic DNA chip for rapid detection and identificaton of retroviruses, retroviral contaminants, and mistaken identity of cell lines.

In: DNA Fingerprinting, Sequencing and Chips. New York, NY: Nova Science Publishers, 2009. 87-118
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Retroviruses not only represent some of the most dangerous pathogens, but also constitute, as remnants of former infections that happened millions of years ago, a large fraction of the human genome. We have established a fast and reliable DNA chip-based assay (RetroArray) for detection and identification of a wide variety of human and other vertebrate exogenous and endogenous retroviruses in biological/clinical samples. The assay combines a pan-retrovirus multiplex polymerase chain reaction (PCR) using fluorochrome-modified primers and DNA chip hybridization. Using RetroArray, distinct transcription profiles of human endogenous retroviruses (HERVS) have been established for a variety of human tissues. Using paired samples (normal vs. Disease) this method can be applied to examine HERV activity in human tumors and can help to identify retrovirus-derived tumor antigens. In addition, RetroArray has been designed to detect human exogenous retroviruses such as human immunodeficity and sensitivity of the assay was demonstrated by detecting traces of pig endogenous retrovirus (PERV) DNA down to ~25 copies in human cDNA samples. Furthermore, retroviral transcripts may be identified in particle preparations from cell culture supernatants. This makes the assay a valuable technique for monitoring packaging cell lines and vector preparations commonly used in human gene therapy applications to exclude cotransfer of replication competent retroviruses (RCRs) or endogenous retrocviruses (ERVs) into target cell. Therefore, RetroArray could improve significantly the safety of human gene therapy, tissue engineering, xenotransplantation and production of therapeutic polypeptides in cell culture. Mistaken identity of human cell lines and frequently observed laboratory contaminations with cells of other species, as well as infection with polytropic animal retroviruses such as squirrel monkey retrocirus (SMRV) or murine leukemia virus (MLV) can influence experimental results and may lead to invalid conclusions. The RetroArray technique is an excellent tool for testing purity and homogeneity of cell lines. Characteristic HERV transcription profiles can be used to assess the cell type and to monitor cell lines for contaminating cells.
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Publikationstyp Artikel: Sammelbandbeitrag/Buchkapitel
ISBN 978-1-60741-814-6
Bandtitel DNA Fingerprinting, Sequencing and Chips
Quellenangaben Band: , Heft: , Seiten: 87-118 Artikelnummer: , Supplement: ,
Verlag Nova Science Publishers
Verlagsort New York, NY