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Optimizing Testing Time for Methaemoglobin Reduction Test in G6PD Screening: A Comparative Study of Monica
Richard Vikpebah Duneeh1,2, Emmanuel Appiah-Intsiful1, Israel Bedzina1
1Department of Medical Laboratory Sciences, School of Health and Allied Sciences, University of Health and Allied Sciences, Ho, Ghana, uhas.edu.gh.
Advances in Hematology
|November 24, 2025
Summary
The optimal Methaemoglobin Reduction Test (MRT) reaction time for diagnosing Glucose-6-phosphate dehydrogenase (G6PD) deficiency is 120 minutes. This finding supports efficient sample management, particularly for pediatric patients requiring G6PD testing.
Area of Science:
- Clinical Diagnostics
- Biochemistry
- Hematology
Background:
- Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a prevalent inherited red blood cell disorder affecting approximately 500 million individuals globally.
- Accurate and efficient diagnosis of G6PD deficiency is crucial for patient management and preventing hemolytic crises.
Purpose of the Study:
- To determine the optimal reaction time for the Methaemoglobin Reduction Test (MRT) in diagnosing G6PD deficiency.
- To compare the diagnostic performance of the Monica Cheesbrough protocol with a modified Asutifi North Hospital protocol.
Main Methods:
- An experimental study involving 643 participants was conducted to compare two MRT protocols at various time points (90, 120, 150, and 180 minutes).
- Intraclass Correlation Coefficient (ICC), Kappa statistics, and McNemar's test were employed to assess reliability, agreement, and differences in G6PD status.
- Data analysis was performed using SPSS version 26.0, with a p-value less than 0.05 considered statistically significant.
Main Results:
- Excellent reliability and agreement (ICC > 0.967, Kappa = 1.000) were achieved from 120 minutes onwards, indicating optimal diagnostic performance.
- Significant differences in G6PD status were observed at 90 minutes (p < 0.001), with diminishing differences at later time points.
- The prevalence of G6PD deficiency was found to be 5.7%, with 4.5% having fully defective and 1.2% having partially defective enzyme activity.
Conclusions:
- The optimal MRT reaction time for diagnosing G6PD deficiency is 120 minutes, ensuring high reliability and agreement between protocols.
- Utilizing the established optimal time allows for accurate diagnosis with potentially reduced sample volumes and reagent usage, enhancing efficiency, especially in pediatric testing.
- These findings contribute to improved diagnostic strategies for G6PD deficiency, optimizing resource management in clinical settings.
