RESUMEN
Purpose: To date, the diagnosis of nontuberculous mycobacteria (NTM) disease primarily relies on clinical symptoms and radiological features. Our objective was to apply a sequence-based analysis method by using partial gene sequencing of heat shock protein 65 (hsp65) to identify NTM species. Patients and Methods: A total of 32 stored isolates obtained from individuals suspected of having pulmonary NTM infection were subjected to solid Ogawa culture. Genomic DNA from each sample was extracted and used in a conventional polymerase chain reaction (PCR) targeting a specific region of hsp65 gene. Identified amplicons from the PCR were then subjected to targeted sequencing. Analysis of the obtained hsp65 sequence was performed using DNA Baser tool. The consensus sequences obtained were compared to references in the GenBank NCBI database to determine NTM species. Results: We identified several important NTM species which posses opportunistic characteristics. M. abscessus and M. chelonae are the most frequent NTM species identified in this study (40.63% and 18.75%, respectively). These two species have the potential to cause significant infections in human, ranging from opportunistic pulmonary infection to localized skin infection. Additionally, pathogenic NTM members of M. fortuitum group (MFG), M. avium, M. intracellulare, M. kansasii, and M. celatum were also found among all identified species. Conclusion: Sequence-based analysis is a promising method for identifying species of NTM. The hsp65 gene has a high discriminatory power to identify opportunistic pathogen NTM species in specimens in Indonesia. Consequently, hsp65 partial gene sequencing is considerable as an alternative and reliable approach for NTM speciation.
RESUMEN
Introduction: Acne vulgaris (AV) is a common and chronic disorder of the pilosebaceous unit and has a multifactorial pathology, including activities of Cutibacterium acnes (C. acnes) and Staphylococcus epidermidis (S. epidermidis). Antibiotic resistance has become a major concern in dermatology daily practice, and the ability of biofilm formation by both bacteria is suggested to increase antibiotic resistance in acne. Purpose: Our aim was to analyze the comparison of antibiotic resistance between biofilm-forming (BF) and non-biofilm-forming (NBF) strains of C. acnes and S. epidermidis towards seven antibiotics commonly used for acne. Methods: This is a cross-sectional analytical study involving 60 patients with AV. Samples were obtained from closed comedones on the forehead using the standardized skin surface biopsy (SSSB) method at the Cosmetic Dermatology Clinic Dr. Hasan Sadikin in Bandung, Indonesia. Isolates were cultured and identified before undergoing the biofilm-forming test using the tissue culture plate method. Antibiotic susceptibility testing for each antibiotic was then performed using the disc diffusion method. Results: The incidence of antibiotic resistance to clindamycin in BF and NBF C. acnes isolates was 54.5% (p=1.00), while in BF and NBF S. epidermidis isolates, it was 54.5% and 45.5% respectively (p=0.67). The incidence of antibiotic resistance to erythromycin and azithromycin in BF and NBF C. acnes isolates was 54.5% and 63.6% respectively (p=1.00), whereas for S. epidermidis BF and NBF isolates, it was 54.5% (p=1.00). There was no resistance observed to tetracycline, doxycycline, levofloxacin, and cotrimoxazole in all groups. Conclusion: There were no significant differences in resistance against seven antibiotics between the C. acnes and S. epidermidis in BF and NBF groups. Furthermore, although statistically not significant, some resistances were observed against clindamycin, erythromycin, and azithromycin. Consequently, the use of these three antibiotics should be judiciously regulated.
