Molecular identification of mycobacterial infections in nonsputum specimens.

Background: Mycobacterial infections can manifest in various anatomical sites, necessitating the analysis of nonsputum specimens for accurate diagnosis. The aim of this study was to identify the molecular cases of mycobacterial infections in nonsputum specimens using polymerase chain reaction based assays and gene sequencing methods. Methods: This observational study examined 161 nonsputum samples that have been stored in the Clinical Microbiology Laboratory at Hasanuddin University Hospital. Samples were analyzed by microscopy and molecular detection methods according to the standard methods at the Clinical Microbiology Laboratory of Hasanuddin University. Descriptive statistics were utilized to summarize patient demographics, infection characteristics, and outcomes. Results: The samples were collected from patients with an average age of 39.82 years. The anatomical sites of specimen collection varied, with musculoskeletal organs and eyes being the most common. Microbiological analysis revealed a predominance of Gram positive bacteria, with polymicrobial morphology observed. Methicillin susceptible Staphylococcus aureus were the most frequently isolated organisms. Acid fast bacilli were detected in 8.1% of samples. Phylogenetic analysis, based on 16S rRNA gene sequencing, revealed similarities between the samples and known mycobacterial species, including Mycobacterium parmense, Mycobacterium lacus, and Mycobacterium dioxanotrophicus. Conclusions: The findings highlight the microbial diversity observed in these infections. The study advocates for comprehensive diagnostic evaluations and targeted testing strategies based on both clinical and laboratory findings. This knowledge can contribute to improved diagnostic accuracy and optimized treatment strategies for mycobacterial infections.

A Streptococcus suis infection causing pneumonia in Indonesia: A case report.

Streptococcus suis (S. suis) is a zoonotic pathogen that causes pneumonia, sepsis, endocarditis, and meningitis. S. suis is primarily found in the upper respiratory tract of pigs. To our knowledge, the first case of S. suis infection has resulted in pneumonia in Indonesia. A 40-year-old woman suffered from shortness of breath last month. The complaint worsened one week ago. She also complained of a productive cough with thick white phlegm. She has a history of late-stage cervical cancer. The patient's vital signs were normal, except for tachypnea. Vesicular breath sounds, no wheezing, and coarse lung crackles were discovered during a physical examination. A chest x-ray showed patchy airspace opacities and interstitial thickening throughout both lungs. The following results were obtained from routine laboratory leukocytosis. Gram stain of the sputum showed a few Gram-positive cocci, mostly in pairs. We confirmed this finding by performing the blood agar, and chocolate agar revealed small α-hemolytic and catalase-negative colonies. The strain was positive for penicillin and ceftriaxone in antimicrobial susceptibility testing. A combination of penicillin and ceftriaxone intravenous was utilized for definitive treatment. After completing a 14-day course of oral antibiotic medication, the patient was discharged. Her symptoms had subsided. This case should remind physicians about the possibility of cancer associated with S. suis infected patient and no clear history of exposure to pigs or other animals.

Real time monitoring of Staphylococcus aureus biofilm sensitivity towards antibiotics with isothermal microcalorimetry.

Biofilm-associated infections with Staphylococcus aureus are difficult to treat even after administration of antibiotics that according to the standard susceptibility assays are effective. Currently, the assays used in the clinical laboratories to determine the sensitivity of S. aureus towards antibiotics are not representing the behaviour of biofilm-associated S. aureus, since these assays are performed on planktonic bacteria. In research settings, microcalorimetry has been used for antibiotic susceptibility studies. Therefore, in this study we investigated if we can use isothermal microcalorimetry to monitor the response of biofilm towards antibiotic treatment in real-time. We developed a reproducible method to generate biofilm in an isothermal microcalorimeter setup. Using this system, the sensitivity of 5 methicillin-sensitive S. aureus (MSSA) and 5 methicillin-resistant S. aureus (MRSA) strains from different genetic lineages were determined towards: flucloxacillin, cefuroxime, cefotaxime, gentamicin, rifampicin, vancomycin, levofloxacin, clindamycin, erythromycin, linezolid, fusidic acid, co-trimoxazole, and doxycycline. In contrast to conventional assays, our calorimetry-based biofilm susceptibility assay showed that S. aureus biofilms, regardless MSSA or MRSA, can survive the exposure to the maximum serum concentration of all tested antibiotics. The only treatment with a single antibiotic showing a significant reduction in biofilm survival was rifampicin, yet in 20% of the strains, emerging antibiotic resistance was observed. Furthermore, the combination of rifampicin with flucloxacillin, vancomycin or levofloxacin was able to prevent S. aureus biofilm from becoming resistant to rifampicin. Isothermal microcalorimetry allows real-time monitoring of the sensitivity of S. aureus biofilms towards antibiotics in a fast and reliable way.

Detection and identification of mycobacteria in sputum from suspected tuberculosis patients.

BACKGROUND: Detection of Tuberculosis agent like nontuberculous mycobacteria (NTM) species by culture and microscopic methods remains difficult and time consuming. A fast and reliable diagnosis of tuberculosis would greatly improve the control of the disease. The purpose of this study is to compare the conventional multiplex PCR and multiplex PCR reverse cross blot hybridization assay to culture method in terms of mycobacteria species detection. FINDINGS: Among the 117 positively cultured samples, nontuberculous mycobacteria (NTM) species were found in 9 samples of multiplex PCR reverse cross blot hybridization assay; compared to only 3 NTM species found in our conventional multiplex PCR, and 13 NTM species were successfully identified among 162 negatively cultured samples compared to only 5 NTM species identification in conventional multiplex PCR results. CONCLUSIONS: The sensitivity of the multiplex PCR reverse cross blot hybridization assay comparing to culture method was 86.03%, the specificity is 35.46%, the positive predictive value was 41.94% and the negative predictive value was 82.41%. For conventional multiplex PCR these values are 81.62%, 38.65%, 41.89%, 79.51%, respectively. Furthermore, in terms of mycobacteria species detection, the conventional multiplex PCR was relatively equal compared to the multiplex PCR reverse cross blot hybridization assay, and to be particularly having no significant discrepant results on the identification of Mycobacteria tuberculosis in both methods.