Biochemistry & Analytical Biochemistry

Why should we analyze analyte fluorescence instead of concentration for multiplex immunoassays?

International Conference on Clinical Chemistry & Laboratory Medicine

October 17-18, 2016 Chicago, USA

Edmond J Breen

Macquarie University, Australia

Posters & Accepted Abstracts: Biochem Anal Biochem

Abstract:

Tissue samples (plasma, saliva, serum or urine) from 169 patients classified as either normal or having one of seven possible diseases (COPD, Mono, Myeloma, Psoriasis, RA, Sepsis and T2D) are analyzed across three 96-well plates for the presences of 37 analytes using cytokine inflammation multiplexed immunoassay panels. Censoring for concentration data caused problems for analysis of low abundant analytes. Using fluorescence analysis over concentration based analysis allowed analysis of these low abundant analytes. Mixed-effects analysis on the resulting fluorescence and concentration responses reveals a combination of censoring and mapping the fluorescence responses to concentration values, through a 5PL curve, changed observed analyte concentrations. Simulation verifies this, by showing a dependence on the mean florescence response and its distribution on the observed analyte concentration levels. Differences from normality, in the fluorescence responses, can lead to differences in concentration estimates and unreliable probabilities for treatment effects. It is seen that when fluorescence responses are normally distributed, probabilities of treatment effects for fluorescence based t-tests has greater statistical power than the same probabilities from concentration based t-tests. We add evidence that the fluorescence response, unlike concentration values, doesn�??t require censoring and it is seen with respect to differential analysis on the fluorescence responses that background correction is not required.

Biography :

Email: ebreen@proteome.org.au