An automated procedure for measuring biotinidase activity in serum

Area of Science:

• Clinical Chemistry
• Enzymology
• Automation in Diagnostics

Background:

• Biotinidase deficiency can cause serious health issues.
• Accurate quantification of biotinidase activity is crucial for diagnosis.
• Manual assays can be time-consuming and less precise.

Purpose of the Study:

• To adapt a manual colorimetric method for biotinidase activity measurement to an automated platform.
• To evaluate the precision, linearity, and comparability of the automated assay.
• To establish an efficient method for population screening and studies.

Main Methods:

• Modification of a manual colorimetric assay for the Technicon AutoAnalyzer II.
• Measurement of N-biotinyl p-aminobenzoate conversion by serum biotinidase.
• Assessment of intra-run and day-to-day precision using quality-control sera.
• Evaluation of assay linearity across clinically relevant ranges (0.2–11.0 U/L).

Main Results:

• The automated procedure demonstrated good precision with intra-run CV of 2.1% and day-to-day CV of 4.6%.
• Results showed linear correlation with biotinidase activity over the 0.2–11.0 U/L range.
• The automated assay results were comparable to the manual method, showing no significant difference.

Conclusions:

• The automated biotinidase activity assay is a reliable and efficient alternative to manual methods.
• The enhanced precision and speed make it ideal for large-scale population studies and newborn screening.
• This automated method facilitates more accessible and accurate diagnosis of biotinidase-related disorders.