Diagnosi parallela automatizzata di infezioni virali trasmissibili per via ematica (HIV, HCV, HBV)

Automated parallel diagnosis of viral infections transmitted in haematic tract. The aim of this study was to develop a complete system for the identification and quantification of HBV, HCV, HIV using molecular biological techniques by means of Real Time PCR. The first step was to develop a positive control, valid for all considered viruses, absolutely free from risk of infection due to the fact that they are synthetic clones. The second step of this study was to develop a complete kit in basic PCR with detection by gel electrophoresis, to identify specific sequences of viral genomes. This test uses a solid thermopolimers mix which allows retrotranscription and amplification to be performed separately in a single vial. The third step was to develop a complete kit in real time PCR to quantify the viral concentrations using, in a first phase, a simple liquid mix and, in a second phase, a solid thermopolimers mix. At the end of my ph.D study it was produced a complete system containing a positive control to check the system and to make a real time standard curve. The real innovation of this study was the development of a solid master mix that delivers a rapid but highly specific test for the 3 viruses simultaneous