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

Journal article

Here we describe the development and validation of a highly sensitive assay of antigen-specific IFN-γ production using real time quantitative PCR (qPCR) for two reporters - monokine-induced by IFN-γ (MIG) and the IFN-γ 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-γ Elispot assay. We observed a clear quantitative correlation between the two assays (Pandlt;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. © 2011 Kasprowicz et al.

Authors: Kasprowicz, V; Mitchell, J; Chetty, S; Govender, P; Huang, K

• Journal: PLoS ONE
• Volume: 6
• Issue: 8
• Publication date: 2011-01-01
• DOI: 10.1371/journal.pone.0020606
• EISSN: 1932-6203
• Language: English