روش حساس كِمي لومينسانس براي سنجش فعاليت آنزيم سوپراكسيد ديسموتاز

Sensitive chemiluminescence method for Superoxide Dismutase activity assay

Abstract Aims: Measuring superoxide dismutase activity is of importance in common diseases like cardiovascular disease, cancer and autoimmune disorders. Nowadays measuring enzyme activity is performed with expensive imported kits in different fields of science. The aim of this study was to design a quantitative method for chemiluminometeric measurement of superoxide dismutase activity with improved sensitivity, accuracy and speed. Materials & Methods: In this method luminol substrate and iodophenol were used as reaction accelerators in luminometry measuring reaction in order to determine the superoxide dismutase activity in serum samples and to increase measurement sensitivity. Sensitivity, precision tests, intra and inter-assay test, accuracy tests, recovery and parallelism tests, method comparison and methodsʹ correlation and coherence investigation were also performed. In order to increase reading speed and precision, measurement was performed in microplate and reading was done in luminometry plate. Results: Standard operating range was 3 to 300 U/ml. Sensitivity of the method was O.lU/ml. Intra and inter- assay coefficient of variation was less than 6.6 and 7.3% respectively. In recovery and Parallelism tests, the recovery percent range was 92 to 110. Comparison of common colorimetric method and the designed method indicated a Pearsonʹs correlation coefficient of 0.917. Conclusion: This method can measure superoxide dismutase activity in serum samples with acceptable precision and accuracy and probably can be a good alternative for conventional colorimetric methods. Keywords: Chemiluminescence, Superoxide Dismutase, Microplate Reading Format