title: PCR-ELISA: a diagnostic assay for identifying Iranian HIV seropositives.
abstract: Quantitative viral load monitoring is an important indicator of prognosis in human immunodeficiency virus type 1 (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.
authors.affiliation: Reproductive Biotechnology Research Center, Avicenna Research Institute, Academic Center for Education, Culture and Research(ACECR), Tehran, Iran. R.Bagheri@avicenna.ac.ir
authors.forename: Rezvan
authors.initials: R
authors.lastname: Bagheri
authors.affiliation: Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran.
authors.forename: Bahareh
authors.initials: B
authors.lastname: Rabbani
authors.affiliation: Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran.
authors.forename: Nejat
authors.initials: N
authors.lastname: Mahdieh
authors.affiliation: Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran.
authors.forename: Hossein
authors.initials: H
authors.lastname: Khanahmad
authors.affiliation: Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran.
authors.forename: Mansour
authors.initials: M
authors.lastname: Abachi
authors.affiliation: Genetic Medical Group, Faculty of Medicine, Ghazvin University of Medical Sciences, Ghazvin, Iran.
authors.forename: Soheila
authors.initials: S
authors.lastname: Asgari
mesh_terms: D004279:DNA, Viral / Q000097:blood / Q000235:genetics / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006679:HIV Seropositivity / Q000097:blood / Q000175:diagnosis / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology; D006801:Humans; D007492:Iran; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019562:Viral Load / Q000235:genetics / Q000276:immunology
chemical_list: D004279:DNA, Viral