Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran ; Department of Molecular Biology and Genetic Engineering, Stem Cell Technology Research Center, No. 9, Saadat abad ave, P.O. Box 19977-75555, Tehran, Iran ; Day General Hospital Laboratory, Tehran, Iran.
Indian J Microbiol. 2012 Sep;52(3):456-63. doi: 10.1007/s12088-012-0271-1. Epub 2012 May 11.
At least 10 million individuals worldwide are co-infected with immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV). These two viruses are transmitted most primarily by exposure to infected blood or blood products. Various nucleic acid assays have been developed for diagnostics and therapeutic monitoring of infections. In the present study, a multiplex real-time PCR assay for simultaneous detection of HCV and HIV-1 using molecular beacons were designed and validated. 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 analysis of scalar concentrations of the samples indicated that this multiplex procedure detects at least 1,000 copies/ml of HIV-1 and 100 copies/ml of HCV with linear reference curve (R (2) > 0.94). The results demonstrate that a specificity of 100 % and sensitivity of 96 % can be achieved. The analytical sensitivity 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 only detected HIV-1 and all major variants of HCV. This assay may represent an alternative rapid and relatively inexpensive screening method for detection of HIV-1/HCV co-infection especially in blood screening.
全球至少有 1000 万人同时感染了艾滋病毒 1 型 (HIV-1) 和丙型肝炎病毒 (HCV)。这两种病毒主要通过接触受感染的血液或血液制品传播。已经开发出各种核酸检测方法来诊断和监测感染情况。在本研究中，设计并验证了一种使用分子信标同时检测 HCV 和 HIV-1 的多重实时 PCR 检测方法。使用 HIV-1 pol 基因和 HCV 基因组 5'NCR 中的保守区域来设计引物和分子信标。对样品的标量浓度分析表明，这种多重方法可以检测到至少 1000 拷贝/ml 的 HIV-1 和 100 拷贝/ml 的 HCV，线性参考曲线 (R (2)> 0.94)。结果表明，可达到 100%的特异性和 96%的灵敏度。使用 BLAST 软件进行的分析敏感性研究表明，引物除 HIV-1 或 HCV 序列外，不会与任何其他序列结合。引物和分子信标探针仅检测到 HIV-1 和 HCV 的所有主要变体。该检测方法可能是一种替代的快速且相对廉价的筛查方法，可用于检测 HIV-1/HCV 合并感染，尤其是在血液筛查中。
