Simultaneous detection of HCV and HIV-1 by SYBR Green real time multiplex RT-PCR technique in plasma samples.
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.
Department of Clinical and Experimental Medicine, Microbiology Section, University of Bologna, St. Orsola Hospital, Via Massarenti 9, 40138 Bologna, Italy. davide.gibellini@unibo.it
Davide
D
Gibellini
Federica
F
Gardini
Francesca
F
Vitone
Pasqua
P
Schiavone
Giuliano
G
Furlini
Maria Carla
MC
Re
D001483:Base Sequence; D052160:Benzothiazoles; D017931:DNA Primers / Q000096:biosynthesis / Q000235:genetics; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D005456:Fluorescent Dyes / Q000737:chemistry; D006679:HIV Seropositivity / Q000097:blood / Q000175:diagnosis / Q000821:virology; D015497:HIV-1 / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D009930:Organic Chemicals / Q000737:chemistry; D011804:Quinolines; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019562:Viral Load
D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I