A user-friendly, highly sensitive assay to detect the IFN-gamma secretion by T cells

作者：Corstjens Paul L A M^*; Zuiderwijk Michel; Tanke Hans J; van der Ploeg van Schip Jolien J; Ottenhoff Tom H M; Geluk Annemiek

来源：Clinical Biochemistry, 2008, 41(6): 440-444.

DOI：10.1016/j.clinbiochem.2007.12.015

• cell mediated immunity
• IFN-gamma
• interferon
• lateral flow
• leprosy
• mycobacteria
• UCP
• up-converting phosphor
• UPT

Objectives: Development of a user-friendly test alternative to ELISA-based assays to detect IFN-gamma by in vitro cultured peripheral blood mononuclear cells (PBMC) stimulated with pathogen-derived antigens.
Design and methods: The molecular components of an operational IFN-gamma ELISA-based test were applied in a lateral flow (LF) immuno-sandwich assay using up-converting phosphor (UCP) reporter particles. The analytical sensitivity of the UCP-LF IFN-gamma assay (ULIGA) was determined and the assay was qualitatively validated with a selection of 60 supernatants derived from PBMC cultures stimulated with M leprae derived antigens, mitogen or medium alone.
Results: ULIGA indicated an analytical sensitivity better than 2 pg/mL, and demonstrated four orders of magnitude dynamic range. The assay correlated well with the IFN-gamma ELISA.
Conclusions: ULIGA allows detection well below the cutoff value (100 pg/mL) used to define positive responses in the IFN-gamma ELISA. The test procedure is less demanding in respect to equipment and labor, and is suited for testing single samples.

• 出版日期2008-4