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- W2280135107 abstract "Abstract ELISpot assays measure antigen specific cell mediated immunity, and are useful indicators of vaccine and therapeutic efficacy. Methods that yield consistent and reproducible results are required to compare data in longitudinal studies or among laboratories. Here, we describe the performance of a validated ELISpot assay using PBMC controls and standardized antigenic stimulants among laboratories participating in an External Quality Assurance (EQA) Program. Responses from four PBMC controls (high, medium, low and negative responders) were compared in an IFN-γ ELISpot assay using SeraCare (SC) kits and individual laboratory specific assay protocol and kits in an ELISpot EQA. SC kit performance was also compared to competitor kits in previous rounds of an EQA, each with 13 participants. SC kits showed high sensitivity in detection of low responders (Spot forming cells [SFC] range 10.9 - 49.2/well) compared with in house kit results (SFC range 0.4 - 67.2). Competitor kits yielded larger numbers of Too Numerous To Count results with high responders and greater variability in comparison to SC kits. SC kits yielded a broad linear dose response curve (10-1000 SFC/well) as a result of smaller spot size and improved resolution. SC kits and standardized assay protocols used by multiple laboratories allowed improved sensitivity of detection of low level antigenic responses, higher consistency and a broad linear range." @default.
- W2280135107 created "2016-06-24" @default.
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- W2280135107 date "2009-04-01" @default.
- W2280135107 modified "2023-09-24" @default.
- W2280135107 title "Consistency and sensitivity of immune response measurements among multiple laboratories using a standardized ELISpot assay kit (42.28)" @default.
- W2280135107 doi "https://doi.org/10.4049/jimmunol.182.supp.42.28" @default.
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