PuSH - Publikationsserver des Helmholtz Zentrums München: Total Internal Reflection (TIRF)-based quantification of procalcitonin for sepsis diagnosis

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Rascher, D. ; Geerlof, A. ; Kremmer, E. ; Krämer, P. ; Michael, S. ; Hartmann, A. ; Rieger, M.

Total Internal Reflection (TIRF)-based quantification of procalcitonin for sepsis diagnosis - a point-of-care testing application.

Biosens. Bioelectron. 59, 251-258 (2014)

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Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.

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A new, highly sensitive fluorescence immunoassay for a TIRF (total internal reflection)-based point-of-care testing (POCT) device was developed for the detection of procalcitonin (PCT), a specific and early marker for sepsis and microbial infections. The immunoassay was performed on a bench-top system that fulfilled all the necessary characteristics of a POCT application, including a short measurement time (<9min), no sample pre-treatment requirements and application directly near patients. New rat monoclonal antibodies targeting PCT were screened and characterized. The best combinations of antibodies were then integrated into single-use cartridges, and the reduction of nonspecific binding was achieved by supplying suitable additives. Moreover, human recombinant PCT (hrPCT) for use as a standard was developed in the native form of hPCT in plasma (PCT1-116, PCT3-116). The assay achieves the required sensitivity range in human plasma to allow reliable differentiation between healthy persons and varying stages of infection severity (LOD=0.04ng/mL; LOQ=0.12ng/mL). Furthermore, the developed PCT assay can be applied in whole human blood with an adequate sensitivity (LOD=0.02ng/mL; LOQ=0.09ng/mL). To the best of our knowledge, this is the first diagnostic test for sepsis to use whole blood, which is a crucial requirement for POCT. We were able to detect native PCT in patient samples and showed a good correlation (R(2)=0.988) with the results of the Kryptor(®) device from BRAHMS, a state of the art device for the detection of PCT.

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