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Die Quantifizierung von proviraler FIV-DNA mittels kompetitiver PCR


Allenspach, K; Amacker, M; Leutenegger, C M; Hottiger, Michael; Hofmann-Lehmann, Regina; Hübscher, Ulrich; Pistello, M; Lutz, Hans (1996). Die Quantifizierung von proviraler FIV-DNA mittels kompetitiver PCR. Schweizer Archiv für Tierheilkunde, 138(2):87-92.

Abstract

Quantification of provirus copies is important in the context of different biological questions. The most reliable approach for DNA quantification is a PCR based on coamplification of two templates of similar length, the target sequence and the reference template, sharing the same primer recognition sequences. During the amplification, the two templates compete for the same primer set (competitive PCR, or cPCR) AND consequently amplify at the same rate independently of the number of cycles. The amplified products can be distingushed by their different lengths. After densitometrical analysis, the proviral copy number of experimentally feline immunodeficiency virus infected cats could be calculated, since a known amount of reference template was used. The method described here proved to be very sensitive (10 copies for the competitor-DNA) and was used to quantitate the proviral load during several experiments in which the influence of periodical immunestimulations and the effect of vaccines on the virus load was studied.

Abstract

Quantification of provirus copies is important in the context of different biological questions. The most reliable approach for DNA quantification is a PCR based on coamplification of two templates of similar length, the target sequence and the reference template, sharing the same primer recognition sequences. During the amplification, the two templates compete for the same primer set (competitive PCR, or cPCR) AND consequently amplify at the same rate independently of the number of cycles. The amplified products can be distingushed by their different lengths. After densitometrical analysis, the proviral copy number of experimentally feline immunodeficiency virus infected cats could be calculated, since a known amount of reference template was used. The method described here proved to be very sensitive (10 copies for the competitor-DNA) and was used to quantitate the proviral load during several experiments in which the influence of periodical immunestimulations and the effect of vaccines on the virus load was studied.

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Additional indexing

Other titles:Quantification of proviral FIV DNA using competitive PCR
Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
630 Agriculture
Uncontrolled Keywords:FIV - proviral load - model for HIV-infection - quantitative competitive PCR
Language:German
Date:1996
Deposited On:28 Dec 2016 10:55
Last Modified:28 Dec 2016 10:55
Publisher:Gesellschaft Schweizer Tierärztinnen und Tierärzte
ISSN:0036-7281
Free access at:Publisher DOI. An embargo period may apply.
Related URLs:http://doi.org/10.5169/seals-589672 (Library Catalogue)
PubMed ID:8720733

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