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Adenosine triphosphate content of Mycobacterium leprae by percoll buoyant density centrifugation
1Department of Dermatology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Abstract:
Thirty nine untreated patients of bacilliferous leprosy with a mean bacteriological index of 4.8 and morphological index of 1.3% formed the study group. Adenosine triphosphate assay was carried out by (i) enzyme treatment method in 18 patients and (ii) percoll buoyant density gradient method in 21 patients. ATP content obtained by percoll buoyant density gradient method was significantly higher than that obtained by enzyme treatment method. Percoll buoyant density centrifugation for purification and isolation of bacilli from human leproma is simplier, quicker and can serve as an alternate method of enzyme treatment.
Insights
The percoll buoyant density gradient method for isolating leprosy bacilli is simpler and quicker than the enzyme treatment method. This technique yields higher adenosine triphosphate (ATP) content, offering a more efficient alternative for leprosy research.
Area of Science:
- Medical Microbiology
- Infectious Diseases
- Biochemistry
Background:
- Leprosy, a chronic infectious disease caused by Mycobacterium leprae, requires accurate methods for bacterial quantification.
- Adenosine triphosphate (ATP) assays are used to assess bacterial viability and load in clinical samples.
- Traditional methods for isolating bacilli from leproma tissue can be time-consuming and complex.
Purpose of the Study:
- To compare the efficacy of two methods for adenosine triphosphate (ATP) assay in leprosy patients.
- To evaluate the percoll buoyant density gradient method against the enzyme treatment method for isolating Mycobacterium leprae.
- To determine a simpler, quicker, and more effective method for bacillary purification and isolation from leproma.
Main Methods:
- Study included 39 untreated patients with bacilliferous leprosy.
- Adenosine triphosphate (ATP) assay was performed using two techniques: enzyme treatment (18 patients) and percoll buoyant density gradient centrifugation (21 patients).
- Bacteriological index (mean 4.8) and morphological index (mean 1.3%) were recorded for the study group.
Main Results:
- The percoll buoyant density gradient method yielded significantly higher ATP content compared to the enzyme treatment method.
- This suggests superior purification and isolation of bacilli using the percoll method.
- The percoll method was found to be simpler and quicker for isolating bacilli from human leproma.
Conclusions:
- Percoll buoyant density centrifugation is a simpler, quicker, and more effective alternative to enzyme treatment for isolating leprosy bacilli.
- This method provides higher ATP content, indicating improved bacterial recovery and viability assessment.
- The findings support the adoption of percoll buoyant density gradient centrifugation in leprosy diagnostics and research.