Fluorimetric assay of D-lactate

Area of Science:

• Biochemistry
• Clinical Chemistry
• Analytical Chemistry

Background:

• D-lactate, a stereoisomer of lactate, is present in human biological fluids.
• Elevated D-lactate levels have been associated with certain metabolic disorders, including diabetes mellitus.
• Accurate and sensitive methods for D-lactate quantification in clinical samples are needed.

Purpose of the Study:

• To develop and validate a novel fluorimetric assay for the quantification of D-lactate in human blood.
• To assess the diagnostic potential of D-lactate levels in distinguishing diabetic patients from healthy individuals.

Main Methods:

• Development of an endpoint enzymatic assay utilizing D-lactate dehydrogenase from Staphylococcus epidermidis.
• Validation of the assay's precision (intrabatch and interbatch coefficients of variance) and limit of detection.
• Analysis of D-lactate concentrations in blood samples from healthy individuals and diabetic patients.

Main Results:

• The developed fluorimetric assay demonstrated good precision with intrabatch and interbatch coefficients of variance of 8.7% and 16.6%, respectively.
• The assay's limit of detection in blood was determined to be 3.73 nmol/ml.
• Significantly higher D-lactate concentrations were observed in diabetic patients (20.0 +/- 1.3 nmol/ml) compared to healthy individuals (11.0 +/- 1.2 nmol/ml) (P < 0.01).

Conclusions:

• A reliable fluorimetric assay for D-lactate in human blood has been successfully developed.
• The assay is sensitive and precise enough for clinical application.
• Elevated blood D-lactate levels are a significant indicator of diabetes mellitus, suggesting its utility as a potential biomarker.