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:: Volume 8, Issue 1 (Spring 2011) ::
Sci J Iran Blood Transfus Organ 2011, 8(1): 42-51 Back to browse issues page
Development of NASBA-ELISA technique for detection of HIV-1 RNA
M. Shahrabi, M. Forouzandeh Dr. , F. Sabahi Dr., M. Paryan
Abstract:   (13969 Views)

  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 µM 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 NASBA-ELISA method for detection of HIV-1 developed.

 

Keywords: Nasba, Elisa, Immunoassay, HIV-1
Full-Text [PDF 336 kb]   (2648 Downloads)    
Type of Study: Research | Subject: Biotechnology
Published: 2013/08/28
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Shahrabi M, Forouzandeh M, Sabahi F, Paryan M. Development of NASBA-ELISA technique for detection of HIV-1 RNA. Sci J Iran Blood Transfus Organ. 2011; 8 (1) :42-51
URL: http://bloodjournal.ir/article-1-462-en.html


Volume 8, Issue 1 (Spring 2011) Back to browse issues page
فصلنامه پژوهشی خون Scientific Journal of Iran Blood Transfus Organ
The Scientific Journal of Iranian Blood Transfusion Organization - Copyright 2006 by IBTO
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