%0 Journal Article
%T Molecular beacon probes¨Cbase multiplex NASBA Real-time for detection of HIV-1 and HCV
%A Samira Mohammadi Yeganeh
%A Mahdi Paryan
%A Siamak Mirab Samiee
%A Vahid Kia
%J Iranian Journal of Microbiology
%D 2012
%I Tehran University of Medical Sciences
%X Background and Objectives: Developed in 1991, nucleic acid sequence-based amplification (NASBA) has been introduced as a rapid molecular diagnostic technique, where it has been shown to give quicker results than PCR, and it can also be more sensitive. This paper describes the development of a molecular beacon-based multiplex NASBA assay for simultaneous detection of HIV-1 and HCV in plasma samples.Materials and Methods: A well-conserved region in the HIV-1 pol gene and 5¡¯-NCR of HCV genome were used for primers and molecular beacon design. The performance features of HCV/HIV-1 multiplex NASBA assay including analytical sensitivity and specificity, clinical sensitivity and clinical specificity were evaluated.Results: The analysis of scalar concentrations of the samples indicated that the limit of quantification of the assay was < 1000 copies/ml for HIV-1 and < 500copies/ml for HCV with 95% confidence interval. Multiplex NASBA assay showed a 98% sensitivity and 100% specificity. The analytical specificity study with BLAST software demonstrated that the primers do not attach to any other sequences except for that of HIV-1 or HCV. The primers and molecular beacon probes detected all HCV genotypes and all major variants of HIV-1.Conclusion: This method may represent a relatively inexpensive isothermal method for detection of HIV-1/HCV co-infection in monitoring of patients.
%K HIV-1
%K HCV
%K Multiplex NASBA assay
%K Molecular beacon
%U http://ijm.tums.ac.ir/index.php/ijm/article/view/443/193