Molecular beacon probes-base multiplex NASBA Real-time for detection of HIV-1 and HCV

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.