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Induction/activation and detection of occult viral agents present in mammalian cells


Schüpbach, J (2001). Induction/activation and detection of occult viral agents present in mammalian cells. In: Brown, F; Lewis, A M; Peden, K; Krause, P. Evolving Scientific and Regulatory Perspectives on Cell Substrates for Vaccine Development. Basel: Karger, 425-437.

Abstract

Occult viral agents in cell cultures may include (i) known viruses present at a concentration too low for detection and (ii) unknown viruses. For maximal safety regarding adventitious agents, a strategy is proposed which involves cellular cloning of candidate cell substrates for vaccines. This will render occult viruses present in either all cells or none, thus greatly facilitating their detection. Known and unknown members of virus families known to establish latency are best detected by molecular methods based on broadly conserved sequences and low stringency conditions. The presence of retrovirus particles, known or unknown, in vaccine harvests can be excluded by the Product-Enhanced Reverse Transcriptase (PERT) assay which detects particle-associated reverse transcriptase with the sensitivity of nucleic acid amplification tests. Latent retroviruses can be activated by stimulation with halogenated pyrimidines and/or azacytidine. Endogenous proviruses with no release of particles are of no concern.

Occult viral agents in cell cultures may include (i) known viruses present at a concentration too low for detection and (ii) unknown viruses. For maximal safety regarding adventitious agents, a strategy is proposed which involves cellular cloning of candidate cell substrates for vaccines. This will render occult viruses present in either all cells or none, thus greatly facilitating their detection. Known and unknown members of virus families known to establish latency are best detected by molecular methods based on broadly conserved sequences and low stringency conditions. The presence of retrovirus particles, known or unknown, in vaccine harvests can be excluded by the Product-Enhanced Reverse Transcriptase (PERT) assay which detects particle-associated reverse transcriptase with the sensitivity of nucleic acid amplification tests. Latent retroviruses can be activated by stimulation with halogenated pyrimidines and/or azacytidine. Endogenous proviruses with no release of particles are of no concern.

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Additional indexing

Contributors:International Association for Biologicals
Item Type:Book Section, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Medical Virology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2001
Deposited On:11 Feb 2008 12:28
Last Modified:05 Apr 2016 12:22
Publisher:Karger
Series Name:Developments in Biologicals
Number:106
ISSN:1424-6074 (P) 1662-2960 (E)
ISBN:978-3-8055-7286-6
Additional Information:Rockville, Md., September 1999
Related URLs:http://content.karger.com/ProdukteDB/produkte.asp?Aktion=showproducts&searchWhat=books&ProduktNr=227216
PubMed ID:11761258

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