With our project we plan:

a)                 to trace both the genetic HIV-1 strains dynamic evolution and epidemic pathways in a geographic area that intersects the official/illegal migration routes.

b)                to investigate both the prevalence and the distribution of either known drug resistance mutations and/or polymorphisms potentially associated to protease, reverse transcriptase, and fusion inhibitors.

c)                 design and set-up of a new genotypic assay for anti-retroviral drugs resistance (protease/reverse transcriptase/fusion inhibitors) that, on the basis of the current HIV-1 epidemiology, is able to detect all the HIV-1 subtypes and CRFs. 

 Each partner involved in the project will be invited to carry out its contribution in a period of three years.

In particular, we expect to follow the scheduled steps, as described below: 

1^st year:

Firstly, each partner will be involved in (a) set-up, (b) standardization and (c) inter-laboratory survey of the genotyping procedures. This will be conducted on a shared panel of reference strains provided by international research agency or by participants. In this manner, both the sensitivity and the efficacy of the methods will be checked on an extensive panel of viral variants from different European geographic areas. 

2^nd year:

In a second place, each co-partner will provide different sets of clinical samples from selected HIV-1 positive populations, in accordance with the following scheme: 

a)                 n°100 retrospective samples: blood clinical samples already characterized through commercial anti-retroviral drug resistance assay. Objective: to compare the results provided by the experimental assay vs. FDA-approved method.

b)                n°50 naïve patient samples: blood clinical samples from patients never experienced with any anti-retroviral therapy. Objective: to evaluate the overall prevalence of “primary drug resistance mutations” to protease, reverse transcriptase, and fusion inhibitors.

c)                 n°50 blood clinical samples from patients with therapeutic/virologic failure in treatment with fusion inhibitors associated HAART since 1-3 months. Objective: to evaluate the overall prevalence of drug resistance mutations associated to fusion inhibitors.

d)                n°10 patients in follow-up: blood clinical samples consecutively drawn, at 1-2 months intervals, for at least three serial times (including baseline), from patients in treatment with HAART including fusion inhibitors. Objective: to evaluate the nucleotide sequences evolution in individual HIV-1 clones from patients under T20 combined regimens.

 3^th year:

 Results evaluation.