Development of sarcosine quantification in urine based on enzyme-coupled colorimetric method for prostate cancer diagnosis
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Date
2018-05-17
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Abstract
An enzyme-coupled colorimetric assay for quantification of urinary sarcosine was developed. The proposed
method is a specific reaction based on hydrogen peroxide (H2O2) formation via sarcosine oxidase (SOX). The
liberated H2O2 reacts with Amplex Red in the presence of horseradish peroxidase (HRP) to produce the redfluorescent
oxidation product, resorufin, which can be measured spectrophotometrically (OD570). The method
was performed in the 96-well microtiter plate. Reaction conditions, such as pH and reaction time were optimized.
At the optimum conditions, the limit of detection (LOD) and quantification (LOQ) were found to be 0.7
and 1 μM, respectively. A good linearity was revealed with a coefficient of 0.990. The assay showed no significant
interference from ascorbic acid, glucose and bilirubin. In addition, it is extremely specific for sarcosine rather
than other amino acids. The determination of sarcosine in human urine displayed high accuracy and good
reproducibility. This method is promising to differentiate prostate cancer patients from healthy subjects according
to urinary sarcosine level. Altogether, this study provides a rapid, simple and specific tool to determine urinary
sarcosine which could be useful for prostate cancer diagnosis.
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Keywords
Colorimetric assay, Sarcosine, Prostate cancer, Sarcosine oxidase, Urine