Ragon Institute of MGH, MIT and Harvard, Harvard Medical School, Boston, Massachusetts, United States of America.
PLoS One. 2011;6(8):e20606. doi: 10.1371/journal.pone.0020606. Epub 2011 Aug 11.
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 (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.
在这里，我们描述了一种使用实时定量 PCR（qPCR）检测两种报告分子——干扰素诱导的单核细胞趋化蛋白-1（MIG）和干扰素诱导蛋白-10（IP10）——的抗原特异性 IFN-γ 产生的高灵敏度检测方法的开发和验证。我们开发并验证了该检测方法，并将其应用于 HIV 共感染患者队列中对 CMV、HIV 和结核分枝杆菌（MTB）特异性反应的检测。我们比较了该检测方法与体外 RD1（ESAT-6 和 CFP-10）特异性 IFN-γ Elispot 检测方法的灵敏度。我们观察到这两种检测方法之间存在明显的定量相关性（P<0.001）。我们的检测方法被证明是一种用于检测 MTB 特异性 T 细胞的敏感检测方法，可在手指穿刺的全血样本（50 uL）上进行，并且不受 HIV 介导的免疫抑制影响。该检测平台在其他临床环境中可能对感染的诊断具有实用性。
