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A Single-tube real-time RT-PCR assay for RNA detection and quantification of genetically diverse HIV including rare HIV-1 non-m groups and HIV-2

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Abstract

The RT-PCR method with real-time fluorescence detection was used for the development of a prototype of a diagnostic kit for reliable diagnosis of genetic variants of RNA of HIV-1 groups M, N, O, and P and RNA of HIV-2 in blood plasma and serum. The kit is tolerant to nucleotide mismatches, provides a broad linear range of concentrations of HIV RNA, 100% analytical specificity, and adequate analytical sensitivity: 42 ME/mL (HIV-1 group M), 45 copies/mL (HIV-2), 92 copies/mL (HIV-1 group O), and 90 copies/mL (HIV-1 group N). The kit provided successful detection and measurement of HIV RNA concentration in all the samples of the international reference panel for HIV-1 genotypes. The results of the test correlate with the results of commercial tests.

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Authors and Affiliations

  1. Institute of Cytology and Genetics, Siberian Branch, Russian Academy of Sciences, pr. Akademika Lavrent’eva 10, Novosibirsk, 630090, Russia

    M. A. Prasolova & M. K. Ivanov

  2. Vector-Best, Koltsovo, ul. Pirogova 2, Novosibirsk oblast, 630559, Russia

    M. A. Prasolova, M. K. Ivanov & E. S. Netesova

  3. Novosibirsk State University, ul. Pirogova 2, Novosibirsk, 630090, Russia

    M. A. Prasolova, M. K. Ivanov & G. M. Dymshits

  4. Vector State Research Center of Virology and Biotechnology, Koltsovo, Novosibirsk oblast, 630559, Russia

    N. M. Gashnikova

  5. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, pr. Akademika Lavrent’eva 8, Novosibirsk, 630090, Russia

    G. M. Dymshits

Authors
  1. M. A. Prasolova
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  2. M. K. Ivanov
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  3. N. M. Gashnikova
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  4. E. S. Netesova
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  5. G. M. Dymshits
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Corresponding author

Correspondence to M. A. Prasolova.

Additional information

Original Russian Text © M.A. Prasolova, M.K. Ivanov, N.M. Gashnikova, E.S. Netesova, G.M. Dymshits, 2014, published in Molekulyarnaya Genetika, Mikrobiologiya i Virusologiya, 2014, No. 4, pp. 20–25.

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Prasolova, M.A., Ivanov, M.K., Gashnikova, N.M. et al. A Single-tube real-time RT-PCR assay for RNA detection and quantification of genetically diverse HIV including rare HIV-1 non-m groups and HIV-2. Mol. Genet. Microbiol. Virol. 29, 179–185 (2014). https://doi.org/10.3103/S0891416814040089

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  • DOI: https://doi.org/10.3103/S0891416814040089

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