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PCR-enhanced reverse transcriptase assays (PERT) are sensitive tools for the detection of retroviruses in biological samples. The adaptation of real-time PCR techniques based on fluorescent probes (F-PERT) has added a reliable quantitative capacity to the assay. In the interest of economy and time, the SYBR Green I-based real-time detection system was used to establish a convenient one-step PERT assay (SG-PERT). This assay can be completed in 2h, is linear over six orders of magnitude and can be used to quantify retroviruses belonging to divergent species, such as the human immunodeficiency virus type 1 (HIV-1), murine leukemia virus (MLV) and prototypic foamy virus (PFV).

Original publication

DOI

10.1016/j.jviromet.2008.10.012

Type

Journal article

Journal

Journal of virological methods

Publication Date

03/2009

Volume

156

Pages

1 - 7

Addresses

Jefferiss Trust Laboratories, Wright-Fleming Institute, Imperial College, London W2 1PG, United Kingdom. m.pizzato@imperial.ac.uk

Keywords

Cell Line, Humans, Retroviridae, HIV-1, Spumavirus, Organic Chemicals, Diamines, Quinolines, RNA, Viral, Fluorescent Dyes, Cell Culture Techniques, Sensitivity and Specificity, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Benzothiazoles