Paper Information

Journal:   THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)   SPRING 2011 , Volume 8 , Number 1 (30); Page(s) 42 To 51.
 
Paper: 

DEVELOPMENT OF NASBA-ELISA TECHNIQUE FOR DETECTION OF HIV-1 RNA

 
Author(s):  SHAHRABI M., FOROUZANDEH M.*, SABAHI F., PARYAN M.
 
* DEPARTMENT OF MEDICAL BIOTECHNOLOGY, FACULTY OF MEDICAL SCIENCES, TARBIAT MODARES UNIVERSITY, TEHRAN, IRAN
 
Abstract: 

Background and Objectives: Viral RNA is the first blood marker which appears in HIV-1 infection. RT-PCR and NASBA are the commonly used techniques for amplification of RNA. NASBA technique provides more advantages over RT-PCR. In this study, an NASBA assay combined with an easy, sensitive enzyme-linked immunosorbent assay (ELISA) was developed for detection of HIV-1 RNA.
Materials and Methods: 11-dig-UTP was added to an NASBA reaction mixture and the RNA amplicons were labeled. Then, the dig-labeled NASBA products hybridized to a biotinylated specific probe in hybridization solution buffer and the hybrids were transferred to a streptoavidin-coated plate. After several washing processes and emission of nonspecific products, the final detection of the captured RNA was done by addition of anti-dig antibody-enzyme conjugate and the substrate.
Results: The results demonstrated that, NASBA is an efficient method for amplification of the target genome. Detection of NASBA products by agarose gel electrophoresis showed a unique 176 bp band corresponding to the specific target. For the detection of NASBA products, an ELISA assay was performed; using 0.01 PM probe, the OD of 1.049
± 0.11 was obtained.
Conclusions: In this study, by using suitable primers and probe a highly sensitive and specific NASBAELISA method for detection of HIV-1 developed.

 
Keyword(s): NASBA, ELISA, IMMUNOASSAY, HIV-1
 
References: 
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