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Detection of mitochondrial respiratory dysfunction in circulating lymphocytes using resazurin
Khaled K Abu-Amero1, Thomas M Bosley
1Department of Genetics, King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia. kamero@kfshrc.edu.sa
Archives of Pathology & Laboratory Medicine
|October 4, 2005
Summary
A new resazurin-based assay offers a rapid, noninvasive method to detect mitochondrial respiratory dysfunction. This technique accurately quantifies mitochondrial activity, aiding in the diagnosis of mitochondrial disorders.
Area of Science:
- Biochemistry
- Cell Biology
- Medical Diagnostics
Background:
- Current methods for detecting mitochondrial respiratory dysfunction are invasive and labor-intensive.
- Existing laboratory techniques often yield conflicting results.
- There is a need for a reliable, noninvasive method to assess mitochondrial function.
Purpose of the Study:
- To develop a rapid, reliable, and noninvasive method for detecting oxidative phosphorylation activity.
- To avoid the need for isolating mitochondrial fractions for analysis.
Main Methods:
- Lymphocytes from patients and controls were incubated with resazurin.
- Mitochondrial inhibition was induced using amiodarone.
- Fluorescence intensity was monitored to calculate mitochondrial respiratory activity.
Main Results:
- Mitochondrial respiratory activity was established in 51 control subjects.
- All 6 patients with mitochondrial syndromes exhibited decreased mitochondrial respiratory activity.
- Patient values ranged from 49% to 71% of control levels (P < .001).
Conclusions:
- The resazurin-based technique is a fast and reproducible method.
- This assay can quantify mitochondrial activity.
- It effectively identifies respiratory functional defects in patients with mitochondrial disorders.
