A Simple and Rapid Method for the Detection of HIV-1/HCV in Co-Infected Patients

  Background: Due to some limitations of serological methods in diagnosis of patients infected with HIV-1 (human immunodeficiency virus) and HCV (hepatitis C virus), it is profoundly important to use molecular methods for the detecting of these infectious agents. However, the most significant problems are the exorbitant cost of these methods and the need of a thermocycler which is an expensive instrument. Objectives: The current research recruits a multiplex Nucleic Acid Sequence Base Amplification (NASBA) in order to simultaneously detect HIV-1 and HCV genomes in patients’ plasma samples. Sensitivity and specificity of this method have been evaluated using clinical samples. Materials and Methods: A multiplex NASBA assay for simultaneous detection of HCV and HIV-1 by the use of specific primers were designed and validated. A well-conserved region in the HIV-1 pol gene and 5’-NCR of HCV genome were used. A total of 40 samples of HIV-1 (20 samples) and HCV (20 samples) were used in the NASBA assay. The specificity and sensitivity of the assay were evaluated. Results: Our results have demonstrated that the primers used in the assay had no interrelation with each other and other possible interfering agents in the assay. The analytical sensitivity of the assay for both HIV-1 and HCV was determined to be 1000 copies/mL and the clinical sensitivity and specificity were 93.3% and 100%, respectively. Conclusions: By exploiting this multiplex NASBA assay, it is possible to detect HIV-1 and HCV infection/co-infection in patients’ plasma with a suitable sensitivity and specificity. Furthermore, due to its simplicity and multiplexing feature, it could be used in limited access laboratories in a cost-effective manner.