Simultaneous detection of HCV and HIV-1 by SYBR Green real time multiplex RT-PCR technique in plasma samples

Molecular and Cellular Probes
Davide GibelliniMaria Carla Re

Abstract

This paper describes the development of a SYBR Green-based multiplex real time RT-PCR for the simultaneous detection of HCV and HIV-1 genomes in plasma samples. Viral genomes were identified in the same sample by their distinctive melting temperature (Tm) which are 81.6 and 86.5 degrees C for HIV-1 gag 142 bp amplicon and HCV 5'-NCR region 226 bp amplicon, respectively. Analysis of known scalar concentrations of reference plasma indicated that the multiplex procedure detects at least 500 copies/ml of both HIV-1 and HCV. In addition, we also assayed HIV-1 and HCV viral load in 30 co-infected patients and in 15 blood donors, confirming the sensitivity and specificity of the assay. This method may represent a useful alternative method for the detection of HIV-1/HCV co-infection, reliable for a rapid and relatively inexpensive screening of blood donors.

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Citations

Jan 22, 2011·World Journal of Gastroenterology : WJG·Sidhartha HazariSrikanta Dash
Dec 25, 2013·Journal of Microbiological Methods·Fatih ŞahinerRamazan Gümral
Sep 13, 2008·Expert Review of Molecular Diagnostics·François RouetEric Nerrienet
Jan 5, 2010·Journal of Virological Methods·Elisa De CrignisDavide Gibellini
Feb 22, 2011·Bioorganic & Medicinal Chemistry·Emanuela BalestrieriBeatrice Macchi
Mar 12, 2008·BioTechniques·Dmitry A KhodakovVladimir M Mikhailovich
May 31, 2016·World Journal of Gastroenterology : WJG·Mohammad IrshadMohammad Ahmad Ansari

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