Pcr-Elisa: A Diagnostic Assay for Identifying Iranian Hiv Seropositives Publisher



Bagheri R¹ ; Rabbani B² ; Mahdieh N³ ; Khanahmad H⁴ ; Abachi M⁵ ; Asgari S⁶ Show Affiliations
Source: Molecular Genetics, Microbiology and Virology Published:2013

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. © 2013 Allerton Press, Inc.