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PCR-ELISA: A diagnostic assay for identifying Iranian HIV seropositives

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Abstract

Quantitative viral load monitoring is an important indicator of prognosis in human immunodeficiency virus type I (HIV-1). PCR-ELISA as a quantitative method is proven to be sensitive and specific for quantification of HIV-1. Extracted DNA of thirty seropositive and twenty seronegative individuals which were confirmed by ELISA and western blot were amplified with digoxigenin-labeled nucleotides; and then in ELISA procedure biotin-labeled probes were hybridized to the PCR products. Diluted PCR products were analysed by electrophoresis and ELISA methods. The observation revealed that combination of nested-PCR and ELISA leads to a sensitive and specific identification of three copies in HIV-infected; the specificity and sensitivity were 95% and 96.7%, respectively. In conclusion, PCR-ELISA was 10 fold more sensitive than nested-PCR. This study developed a high sensitive PCR-ELISA to assess the quantification of proviral DNA load in the most relevant case of HIV-1 subtype. The reproducibility and reliability of this high-throughput test makes it appropriate for general laboratories to use for quantification of viral load and clinical diagnosis.

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Authors and Affiliations

  1. Reproductive Biotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research (ACECR), Tehran, Iran

    Rezvan Bagheri

  2. Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran

    Bahareh Rabbani

  3. Medical Genetic Group, Faculty of Medicine, Ham University of Medical Sciences, Ham, Iran

    Nejat Mahdieh

  4. Department of anatomical sciences and molecular biology, Isfahan University of Medical Sciences, Isfahan, Iran

    Hossein Khanahmad

  5. Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

    Mansour Abachi

  6. International campus, Tehran University of Medical Sciences, Tehran, Iran

    Soheila Asgari

Authors
  1. Rezvan Bagheri
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  2. Bahareh Rabbani
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  3. Nejat Mahdieh
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  4. Hossein Khanahmad
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  5. Mansour Abachi
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  6. Soheila Asgari
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Correspondence to Rezvan Bagheri.

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Bagheri, R., Rabbani, B., Mahdieh, N. et al. PCR-ELISA: A diagnostic assay for identifying Iranian HIV seropositives. Mol. Genet. Microbiol. Virol. 28, 127–131 (2013). https://doi.org/10.3103/S0891416813030026

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