Current molecular methods for the detection of GB virus C


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Abstract

The GB virus C (GBV-C), also referred to as hepatitis G virus, is a RNA hepatitis virus, which infects humans naturally and can cause high levels of viremia, which, however, in most cases, does not lead to serious illness. It is not usually routinely screened in clinical laboratories, though its presence in patients infected with other viruses such as hepatitis C virus and HIV has been commonly reported. The virus can be detected in the blood of infected individuals using reverse-transcription polymerase chain reaction (RT-PCR) mainly. It is a technique that is commonly used for different RNA material detection and other screening such as gene expression and testing. The method includes several steps, that is, transcription of the RNA into complementary DNA (cDNA) by reverse transcriptase, amplification of the cDNA by PCR using well defined primers that target a particular region of the gene, and detection of the PCR products by various means such as agarose gel electrophoresis or hybridization and immunoassay using, for example, automated detection systems. Several primers targeting different regions of the genome, that is, NS3, NS5, and 5’ NTR, have been developed for the amplification of the specific region and the detection of the virus. Moreover, variants of the RT-PCR have been designed to maximize the identification potential. These include, for example, RT-PCR-ELISA, RT digital-PCR, and SYBR green real-time multiplex RT-PCR. Another type of method used to screen the virus is the detection of the presence of antibodies E2 (anti-E2) produced against the viral glycoprotein E2 in individuals who have been infected with the virus and have recovered or are in the process of recovering. The anti-E2s are detected by radio-immunoprecipitation assay and ELISA. Use of both antibody and RNA detection methods is necessary for a better epidemiological investigation of the prevalence of GBV-C.

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