A Molecular Assay for Sensitive Detection of Pathogen-Specific T-Cells

作者：Kasprowicz Victoria O^*; Mitchell Jessica E; Chetty Shivan; Govender Pamla; Huang Kuan Hsiang Gary; Fletcher Helen A; Webster Daniel P; Brown Sebastian; Kasmar Anne; Millington Kerry; Day Cheryl L; Mkhwanazi Nompumelelo; McClurg Cheryl; Chonco Fundisiwe; Lalvani Ajit; Walker Bruce D; Ndung'u Thumbi; Klenerman Paul

来源：PLos One, 2011, 6(8): e20606.

DOI：10.1371/journal.pone.0020606

Here we describe the development and validation of a highly sensitive assay of antigen-specific IFN-gamma production using real time quantitative PCR (qPCR) for two reporters monokine-induced by IFN-gamma (MIG) and the IFN-gamma 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-gamma 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.

• 出版日期2011-8-11