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

作者：Samira Mohammadi Yeganeh; Mahdi Paryan; Siamak Mirab Samiee; Vahid Kia

来源：Iranian Journal of Microbiology, 2012.

• HIV-1
• HCV
• Multiplex NASBA assay
• Molecular beacon

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 %26lt; 1000 copies/ml for HIV-1 and %26lt; 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.