PCR sterilization: an evaluation of two methods.

Important note: Information in this article was accurate in 1992. The state of the art may have changed since the publication date.

PCR sterilization: an evaluation of two methods.

Abstr Annu Meet Am Soc Microbiol. 1991 May 5-9;91:335 (abstract no. T-6). Unique Identifier : AIDSLINE ASM91/0190292
Frelow MM; Louie P; Rodgers G; Madej R; Roche Diagnostics Research, 1145 Atlantic Ave., Alameda, CA; 94501.

Abstract: Contamination of polymerase chain reaction (PCR) reactions and DNA samples with previously amplified DNA products (amplicon) has become a source of concern. PCR sterilization involves the modification of elimination of amplicon contaminants. We have evaluated two methods for PCR sterilization: a post-PCR sterilization of the amplicon (HRI) and a pre-PCR sterilization of reactions components (Cetus). In the post-PCR method, amplification is done in the presence of an isopsoralen compound which derivatizes the DNA and renders it unsuitable for subsequent amplification. The pre-PCR method involves enzymatic cleavage by uracil-N-glycosidase (UNG) of dUTP containing amplicon contaminants prior to amplification. Using an oligomer hybridization assay to detect HIV amplification product, both methods were evaluated to determine the maximum amount of contamination that could be eliminated. HIV negative samples were intentionally spiked with various amounts of HIV amplicon. These samples were then amplified for the HIV target and the products assayed to determine sterilization efficiency. The two methods produced similar sterilization efficiencies.

Keywords: Disinfection DNA, Viral/*ANALYSIS Human HIV/*GENETICS Nucleic Acid Hybridization Nucleosidases/METABOLISM *Polymerase Chain Reaction ABSTRACT
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