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

被引:38
作者
Gibellini, Davide [1 ]
Gardini, Federica [1 ]
Vitone, Francesca [1 ]
Schiavone, Pasqua [1 ]
Furlini, Giuliano [1 ]
Re, Maria Carla [1 ]
机构
[1] Univ Bologna, St Orsola Hosp, Dept Clin & Expt Med, Microbiol Sect, I-40138 Bologna, Italy
关键词
HIV-1; HCV; SYBR green; multiplex RT-PCR;
D O I
10.1016/j.mcp.2005.12.005
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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/nil 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. (c) 2006 Elsevier Ltd. All rights reserved.
引用
收藏
页码:223 / 229
页数:7
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