Abstract
A highly sensitive enzymatic cycling method was developed for the serum total bile acids assay. We constructed a prokaryotic expression system to prepare the recombinant 3α-hydroxysteroid dehydrogenase in place of the natural enzyme and for the first time used it in the total bile acids assay. The production rate of thio-NADH correlated with the bile acids concentration and was measured by the change of absorbance at 405/660 nm. The enzymatic cycling method could detect 0.22 μmol/L total bile acids in serum. Within-run and between-run imprecisions were 1.2–3.7% and 2.3–4.8%, respectively. The calibration curve for total bile acids in serum was linear between 0.5 and 180 μmol/L. This method was free from interference by bilirubin, hemoglobin, ascorbate, and lactate dehydrogenase. In conclusion, serum total bile acids could be measured by the enzymatic cycling method with recombinant 3α-hydroxysteroid dehydrogenase as the tool enzyme.
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