Simultaneous Diagnosis of HIV-1 and HCV Infections by Nucleic Acid Sequence-Based Amplification

Due to some problems of serological assays in diagnosis of patients infected with HIV-1 and HCV, the use of molecular assay has become widely accepted. However, molecular assays have a major drawback of high cost and dependency to expensive thermocycler instruments. This research described the development of a multiplex nucleic acid sequence-based amplification (NASBA) assay for the simultaneous or single detection of HIV-1 and HCV genomes in plasma samples. The sensitivity and specificity of this assay has been evaluated using several clinical samples. The results showed that the primers used in this assay did not have any interactions with each other or with other possible interfering agents. The analytical sensitivity of this assay for both HIV-1 and HCV was calculated as 1000 copies/ml. The clinical sensitivity and specificity of the assay were considered as 90% and 100%, respectively. The developed multiplex NASBA assay can be used for simultaneous or single detection of HIV-1 and HCV viruses with a suitable sensitivity and specificity. Due to easy application, multiplexity, and no need to thermocycler instruments, this assay can be used as a low cost method in laboratories with limited instruments.