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Molecular Identification of Mycobacterium leprae in the Leprosy Patients
Utma Laela Warka1, Mochammad Hatta1,2, Lisa Tenriesa Muslich1,2,3
1Department of Microbiology, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia.
Background:
Several discoveries about leprosy indicate that Mycobacterium leprae transmission mainly occurs by inhalation, and the nose is a major port of entry and exit. Molecular probes have shown certain potential for the detection and identification of M. leprae in patients. The aim of this study was to identify M. leprae in nasal swab specimens using polymerase chain reaction (PCR)-based assays followed by gene sequencing methods. This observational study examines 64 anterior nasal swab samples taken from pretreatment leprosy patients, on-treatment and completed leprosy treatment in Bulukumba, South Sulawesi, Indonesia.
Methods:
samples were analyzed by molecular detection methods according to the standard methods at the Clinical Microbiology Laboratory of Hasanuddin University. Descriptive statistics were utilized to summarize patient demographics and outcomes.
Results:
This study uses PCR to detect the M. leprae deoxyribonucleic acid (DNA) from nasal swab specimens. Data were collected from 64 patients with a percentage of male patients 51.54%. Based on the age category, the group 45-46 years was the most frequent (39.05%). PCR detection proline-rich antigen gene of a 531 bp DNA fragment from M. leprae, was positive in eight patients, and they were multibacillary. Furthermore, PCR was positive in 5 (31.25%) of 16 new leprosy patients, 2 (8.69%) of 23 on-treatment patients, and 1 (4%) of 25 treatment completed patients. Based on the results of the phylogenetic tree and analysis of 8 positive results detected by M. leprae from leprosy patients, almost all samples have a level of similarity, except for sample Ua7.
Conclusions:
M. leprae cannot grow in vitro, so molecular diagnostic tools were used to confirm the disease. This study predominantly of males with the age above 45 years of age being the most common. Eight M. leprae were positive from nasal swab leprosy patients. The sequencing findings provide insight into the genetic diversity of the genus M. leprae, so it is necessary to consider the detection of whole-genome sequence.
Insights
This study detected Mycobacterium leprae (M. leprae) DNA in nasal swabs from leprosy patients using PCR. Eight positive cases were identified, highlighting the nose as a key site for M. leprae detection.
Area of Science:
- Microbiology
- Genetics
- Infectious Diseases
Background:
- Leprosy transmission primarily occurs via inhalation, with the nose serving as a crucial entry and exit point for Mycobacterium leprae (M. leprae).
- Molecular probes show promise for detecting and identifying M. leprae in patients.
Purpose of the Study:
- To identify M. leprae in nasal swab specimens using polymerase chain reaction (PCR)-based assays and gene sequencing.
- To investigate the presence of M. leprae in nasal swabs of leprosy patients at different treatment stages.
Main Methods:
- Analysis of 64 anterior nasal swab samples from leprosy patients (pretreatment, on-treatment, and completed treatment).
- Utilized PCR to detect the M. leprae proline-rich antigen gene (531 bp DNA fragment).
- Phylogenetic analysis of positive M. leprae samples.
Main Results:
- PCR detected M. leprae DNA in 8 out of 64 nasal swab samples, all from multibacillary leprosy patients.
- Positive PCR results were observed in 31.25% of new patients, 8.69% of on-treatment patients, and 4% of completed treatment patients.
- Phylogenetic analysis indicated high similarity among the detected M. leprae samples, with one exception.
Conclusions:
- Molecular diagnostic tools like PCR are essential for confirming M. leprae infection due to the inability to culture the bacteria in vitro.
- The study identified M. leprae in nasal swabs of leprosy patients, predominantly males over 45 years old.
- Sequencing data offer insights into M. leprae genetic diversity, suggesting the need for whole-genome sequencing for comprehensive analysis.
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