Washing-Free and Label-Free Onsite Assay for Inorganic Pyrophosphatase Activity Using a Personal Glucose Meter

Area of Science:

• Biochemistry
• Analytical Chemistry
• Biomedical Diagnostics

Background:

• Inorganic pyrophosphatase (PPase) plays a role in various biological processes.
• Accurate and accessible detection methods for PPase are crucial for diagnostics.
• Existing PPase detection methods often require complex procedures and equipment.

Purpose of the Study:

• To develop a simple, washing-free, and label-free method for inorganic pyrophosphatase (PPase) detection.
• To utilize a personal glucose meter for quantitative PPase activity measurement.
• To validate the method's diagnostic capability using human serum samples.

Main Methods:

• A cascade enzymatic reaction (CER) involving hexokinase and pyruvate kinase was employed.
• The method relies on the interplay between adenosine triphosphate sulfurylase and PPase activity.
• Pyrophosphate (PPi) levels are indirectly measured by monitoring glucose reduction via CER.

Main Results:

• A quantitative PPase detection method was established with a linear dynamic range of 1.5-25 mU/mL.
• The limit of detection for PPase was determined to be 1.18 mU/mL.
• The method demonstrated successful PPase detection in undiluted human serum with 100 ± 2% recovery.

Conclusions:

• The developed method offers a cost-effective and user-friendly alternative for PPase detection.
• The assay eliminates the need for specialized equipment and washing steps.
• This approach holds promise for point-of-care diagnostics and biochemical analysis.