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• W2025516091 endingPage "387" @default.
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• W2025516091 abstract "A quantitative real-time PCR (qPCR) assay using SYBR Green dye was established in order to detect and quantify the proviral DNA of HTLV-1 in peripheral blood mononuclear cells (PBMCs). Primers were designed, and the assay was standardized to amplify a novel, conserved HTLV-1 tax region. Proviral load was normalized to the amount of cellular DNA by quantitation of the human albumin gene. Firstly, the qPCR was assessed determining the specificity, sensitivity, dynamic range and intra- and inter-assay reproducibility of the technique. The limit of detection as determined by PROBIT analysis using dilutions of the standard was 2.97 copies. The assay had an excellent dynamic range from 105 to 101 copies per reaction and good intra- and inter-assay reproducibility, CVs less than 2%. Secondly, the performance of the qPCR was tested on 40 HTLV-1 seropositive individuals. Proviral load for HTLV-1 carriers ranged from 2.2 × 102 to more than 8.3 × 104 copies/106 PBMCs. The high sensitivity and wide dynamic range allowed the determination of a broad range of HTLV-1 proviral loads in infected individuals. This assay is a valuable alternative diagnostic tool when current available serological assays are insufficient. In addition, it will facilitate the study of the relationship between proviral load and pathogenesis." @default.
• W2025516091 created "2016-06-24" @default.
• W2025516091 creator A5009254139 @default.
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• W2025516091 date "2013-05-01" @default.
• W2025516091 modified "2023-09-30" @default.
• W2025516091 title "Development and validation of a real-time PCR assay for a novel HTLV-1 tax sequence detection and proviral load quantitation" @default.
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• W2025516091 doi "https://doi.org/10.1016/j.jviromet.2013.02.018" @default.
• W2025516091 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/23499262" @default.
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