A molecular assay for sensitive detection of pathogen-specific T-cells.

ABSTRACT: Here we describe the development and validation of a highly sensitive assay of antigen-specific IFN-? production using real time quantitative PCR (qPCR) for two reporters--monokine-induced by IFN-? (MIG) and the IFN-? inducible protein-10 (IP10). We developed and validated the assay and applied it to the detection of CMV, HIV and Mycobacterium tuberculosis (MTB) specific responses, in a cohort of HIV co-infected patients. We compared the sensitivity of this assay to that of the ex vivo RD1 (ESAT-6 and CFP-10)-specific IFN-? Elispot assay. We observed a clear quantitative correlation between the two assays (P<0.001). Our assay proved to be a sensitive assay for the detection of MTB-specific T cells, could be performed on whole blood samples of fingerprick (50 uL) volumes, and was not affected by HIV-mediated immunosuppression. This assay platform is potentially of utility in diagnosis of infection in this and other clinical settings.

SUBMITTER: Kasprowicz VO 

PROVIDER: S-EPMC3154901 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature