Validation of the ELISA Method for Quantitative Detection of TNF-Î± in Patients with Intracerebral Hemorrhage
Keywords:TNF-Î±, intracerebral haemorrhage, ELISA, standard addition, spectroscopy
AIM: We aimed to investigate the sensitivity, reproducibility and validity of the commercial ELISA kits for quantitative detection of TNF-Î± and their potential application for screening purposes in patients with ICH.
METHODS: Analysis of six independent standard series, evaluation of the deviation of the TNF-Î± concentration in patients with ICH, standard addition and visual analysis of whole UV-Vis spectra were carefully performed.
RESULTS: Low standard deviations of the absorbance were detected for every standard, as well as in the samples of healthy controls and patients with ICH. The standard addition series have also confirmed high sensitivity and reproducibility of the assay, with a congruent shift of the standard curves with the concentration of TNF-Î± for the added plasma. The visual analyses of the gained spectra have revealed the absence of any matrix effects from the addition of the human plasma in the reconstituted standards.CONCLUSION: The commercial ELISA kits can be used in the clinical practice for screening purposes of the plasma TNF-Î± levels in patients with ICH.
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