Final Activity Report Summary - BEACON (Novel molecular beacon technologies for the study of hiv-1 and other infectious agents)
The original scope of the project was to investigate the use of molecular beacon technologies in other infectious agents aiming to develop a unique approach for a class of challenging problems in human genetics and infectious diseases. Molecular beacons are synthetic hairpin-shaped nucleic acid probes that undergo a fluorogenic conformational change upon binding to their nucleotide-specific target. As a consequence of their unique architecture, they are extraordinarily specific even in the presence of genetically quasi-similar targets. A single nucleotide mismatch between the probe sequence and the target sequence prevents hybridization. Molecular beacons can be labelled with differently coloured fluorophores, enabling multiplex, homogeneous amplification assays to be carried out in real-time in sealed reaction tubes. Among the applications are the detection of infectious viral and microbial agents in clinical and environmental samples, the quantitative assessment of gene expression, and the determination of genotypes in human alleles associated with human disease.
The proposal dealt with the investigation of 1) the role of the concentration of HIV-1 DNA in peripheral blood mononuclear cells (cellular viral load), in predicting the virological response to treatment 2) the determination if single-dose neviparine (NVP) used to prevent HIV-1 perinatal transmission is associated with the selection of a higher frequency of resistance mutations in HIV-1 infected mothers and 3) the HIV-1 genetic diversity and determination of the molecular epidemiology and spread of drug resistance of HIV-1 infection in Cyprus.
Concerning the overall aims set in the proposal, we have succeded in establishing a national infrastructure in collaboration with the AIDS Department of the Ministry of Health and the Gregorian AIDS Clinic of well trained, a quality controlled reference HIV virology laboratory in the University of Cyprus for studies associated with the HIV molecular epidemiology and genotypic resistance in Cyprus.
We have also managed to enrol Cyprus in the European network, EuropeHIVResistance, which aims to implement a European strategy to prevent transmission of resistant HIV-1 strains in close collaboration with a network of the Associated States of Europe. In addition we have developed a Cypriot database and viral panels of HIV-1 strains, which may be used for development of specialized diagnostic assays, HIV antiretroviral compounds and vaccines. Furthermore, genotypic drug resistance analyses were performed on drug-naive patients in the course of molecular analyses of the Protease and Reverse Transcriptase genes.
The proposal dealt with the investigation of 1) the role of the concentration of HIV-1 DNA in peripheral blood mononuclear cells (cellular viral load), in predicting the virological response to treatment 2) the determination if single-dose neviparine (NVP) used to prevent HIV-1 perinatal transmission is associated with the selection of a higher frequency of resistance mutations in HIV-1 infected mothers and 3) the HIV-1 genetic diversity and determination of the molecular epidemiology and spread of drug resistance of HIV-1 infection in Cyprus.
Concerning the overall aims set in the proposal, we have succeded in establishing a national infrastructure in collaboration with the AIDS Department of the Ministry of Health and the Gregorian AIDS Clinic of well trained, a quality controlled reference HIV virology laboratory in the University of Cyprus for studies associated with the HIV molecular epidemiology and genotypic resistance in Cyprus.
We have also managed to enrol Cyprus in the European network, EuropeHIVResistance, which aims to implement a European strategy to prevent transmission of resistant HIV-1 strains in close collaboration with a network of the Associated States of Europe. In addition we have developed a Cypriot database and viral panels of HIV-1 strains, which may be used for development of specialized diagnostic assays, HIV antiretroviral compounds and vaccines. Furthermore, genotypic drug resistance analyses were performed on drug-naive patients in the course of molecular analyses of the Protease and Reverse Transcriptase genes.