Jundishapur Journal of Microbiology
Volume:14 Issue: 7, Jul 2021

Indonesia is one of the five countries with the highest number of diphtheria cases worldwide. Diphtheria is caused by the toxigenic strains Corynebacterium diphtheriae, C. ulcerans, and C. pseudotuberculosis. The diphtheria-causing bacteria can be identified using conventional and molecular methods, including polymerase chain reaction (PCR) assay. We used the PCR assay as additional testing, because in island countries like Indonesia, specimen transport is a serious challenge to maintaining bacterial survival.

This study aimed to evaluate the PCR assay as additional testing to identify diphtheria-causing bacteria in the diphtheria laboratory.

In this cross-sectional study, a total of 178 pairs of the throat and nasal swabs from diphtheria suspected cases and close contacts were collected from seven provinces in Indonesia in 2016. All samples were directly identified by the conventional method and multiplex PCR assay. Statistical analysis was conducted using the 2 × 2 tables to determine the sensitivity and specificity of both methods, while the χ 2 test was used to examine the correlation between specimen examination delay and the differentiation of results. A P-value < 0.05 was considered as statistically significant.

Out of 178 examined samples, eight samples were identified as diphtheria-positive by both the conventional method and PCR assay, while nine samples were only detected by the PCR assay. All diphtheria-causing bacteria found in the positive samples were toxigenic C. diphtheriae. The diphtheria-causing bacteria were found in 27.6% of cases and 6.0% of close contacts using the PCR assay versus 13.8% of cases and 2.7% of close contacts using the conventional method. Statistical analysis showed that the PCR assay is about twice more sensitive than the conventional method. There was a significant correlation between the differentiation of results and > 72 hours’ specimen examination delay with a P-value of 0.04 (< 0.05).

The PCR assay is more sensitive than the conventional method to identify diphtheria-causing bacteria and may be applied as additional testing to increase the positivity rate of diphtheria-confirmed cases in Indonesia as an archipelago country where geographical factors and specimen transport are real obstacles.

The prevalence of nontuberculous mycobacteria (NTM) infection has been increasing globally. Many cases of NTM infection are misdiagnosed as Mycobacterium tuberculous (MTB) because of similar clinicoradiological features.

To determine the burden and characteristics of NTM infection, this study was done to evaluate clinical isolates collected from tuberculous (TB) suspects in a population from Northwest China.

From January to December 2020, the clinical samples of 9,142 TB suspects were collected for the PCR-fluorescent probe and mycobacterial culture. The PCR-fluorescent probe-positive nucleic acid samples were further subjected to a DNA microarray for confirmation and species identification. Drug susceptibility testing (DST) was also carried out using the micropore plate method (MicroDSTTM) on isolates from NTM patients.

Of 9,412 TB suspects, 85 cases (0.9%) were clinically diagnosed with NTM infection according to the American Thoracic Society (ATS) guidelines. For the laboratory samples, a total of 169 NTM strains, identified by molecular biology methods, were classified into 10 species. Themost common species were M. chelonae/ M. abscessus (64/169, 37.7%) and M. intracellulare (40/169, 23.7%). All strains showed the highest resistance to imipenem/cilastatin (85/85, 100%) and the highest susceptibility to linezolid (4/85, 4.7%). In comparison with the rapidly growing mycobacteria (RGM) group, the slowly growing mycobacteria (SGM) group showed a lower resistance and a shorter hospital inpatient stay (t = 6.66, P < 0.001 and t = 2.40, P = 0.020, respectively).

Mycobacterium chelonae/M. abscessus and M. intracellulare were the most frequently detected NTM pathogens in Northwest China. The differences in drug sensitivity and clinical characteristics were giant for different strains. Timely identification and accurate DST play important roles in NTM management.

Pseudomonas aeruginosa is a unique Gram-negative opportunistic pathogen that is the leading cause of nosocomial infections.

This study aimed to investigate the prevalence of the main carbapenemase and extended-spectrum β-lactamases encoding genes in P. aeruginosa clinical isolates.

In the present study, we collected 85 P. aeruginosa clinical isolates from different wards of three military hospitals in Tehran, Iran. We used disk diffusion and agar dilution methods to determine resistance to 12 different antibiotics in these isolates. Also, we assessed the blaIMP, blaVIM, blaSHV , blaTEM, and blaCTX genes by polymerase chain reaction methods among all isolates.

Our results revealed that all isolates were resistant to two antibiotics, and 76 (89.4%) of isolates were multidrug-resistant. We observed maximum and minimum resistance rates against ticarcillin (n = 77; 90.5%) and colistin (n = 7; 8.2%), respectively. The blaVIM, blaIPM, blaTEM, blaSHV , and blaCTX genes were harbored by 44 (51.8%), 20 (23.5%), 41 (48.2%), 24 (28.2%), and 16 (18.8%) isolates, respectively.

The resistance rate among P. aeruginosa strains is significantly increasing that causes nosocomial infections due to different mechanisms, including the high frequency of metallo-β-lactamases and extended-spectrum β-lactamases genes.

Staphylococcus aureus is an important cause of resistant infection with high mortality and morbidity.

We aimed to evaluate the clinical characteristics and comorbidities of patients with S. aureus infection to define the predictors of adverse outcomes.

In this cross-sectional study, patients (aged ≥ 15 years) with positive S. aureus blood cultures were included. Their demographic and clinical characteristics were recorded, and their association with the main adverse outcomes (methicillin-resistant S. aureus [MRSA], infective endocarditis, source of infection, and the final outcome were analyzed using SPSS software version 16.

The male-to-female ratio was 54/51. The mean age was 55.13 years (women: 58.45 ± 20.4 and men: 53.6 ± 17.6). Of 105 cases analyzed, 40% had hospital-, 25.7% community-, and 34.3% healthcare-associated bacteremia. The median duration of hospital admission was 13 days. Thirty-two percent had MRSA, differently based on the source of infection (P = 0.029). Twenty-eight patients had infective endocarditis, differently based on the source of infection, prosthetics, considerable foci of infection, and receipt of blood and its derivatives (P < 0.05). Most patients with neurological and end-stage renal disease (both P = 0.001) did not have infective endocarditis. Finally, 61.9% of the patients were discharged with good condition, 38.1% died, and 9% left the hospital before a definite diagnosis.

Vascular catheters and cardiovascular diseases, including hypertension, are among the most common factors associated with S. aureus bacteremia, and it is necessary to carefully examine the presence of these factors, as well as infective endocarditis in these patients.

Candidemia is the most common systemic infection in hospitalized patients causing high mortality. Hence, the diagnosis of this infection in the early stage with appropriate antifungal therapy is paramount.

The study aimed at molecular identification of Candida species isolated from candidemia patients and evaluation of the in vitro antifungal susceptibility patterns of these strains to fluconazole, amphotericin B, and caspofungin.

In the present study, 800 hospitalized patients who were suspected to have candidemia were sampled. Candida species were isolated and identified based on morphological characteristics and PCR-sequencing of the ITS1-5.8S-ITS2 region. Antifungal susceptibility tests for fluconazole, amphotericin B, and caspofungin were performed according to the Clinical and Laboratory Standards Institute protocol M27-A3. Also, clinical data were recorded from the patients’ records.

Twenty-seven patients among the sample of hospitalized patients were found to have candidemia. A total of 33.3% of candidemia patients were treated with amphotericin B, in which case the mortality rate was 14.8%. The majority of patients (59%) were from the neonatal intensive care unit, and premature birth was the most common underlying condition. Candida albicans (n = 18; 66.6%) was the most common species isolated from blood cultures, followed by C. parapsilosis (n = 7; 25.9%), C. pelliculosa (n = 1; 3.7%), and C. tropicalis (n = 1; 3.7%). Only one C. albicans isolate resistant to fluconazole (minimum inhibitory concentration = 32 µg/mL).

Generally, C. albicans has been the most frequent causative agent of candidemia. Resistance to antifungal drugs among candidemia agents was rare. Also, the identification of Candida isolates at the species level with in vitro antifungal susceptibility tests helps manage candidemia patients better and decrease the mortality rate among them.

Fast, reliable, and cost-effective tests are recommended for tuberculosis diagnosis and drug susceptibility testing, especially in resource-limited settings.

This study aimed to evaluate the performance of thin-layer agar for tuberculosis diagnosis and drug susceptibility testing.

Samples were collected from patients with presumptive tuberculosis and tested using thin-layer agar for tuberculosis and drug susceptibility testing in parallel with Lowenstein Jensen culture method for tuberculosis diagnosis and proportion method for drug susceptibility testing as the gold standard. Receiver operating characteristic curve analysis was performed to calculate the performance parameters.

Thin-layer agar method showed sensitivity and specificity values of 96.63% and 62.50%, respectively, for the isolation of Mycobacterium tuberculosis directly from specimens. Drug susceptibility results using thin-layer agar showed sensitivity values for isoniazid, rifampicin), ethambutol and streptomycin were 94.74%, 86.84%, 94.74% and 81.58%, respectively, while the specificity values were 100%, 100%, 86.27% and 100% for isoniazid, rifampicin, ethambutol and streptomycin, respectively. Results were available in a median time of 16 days for thin-layer agar and 25 days for the conventional method.

The thin-layer agarmethod is a relatively rapid, simple, and cost-effectivemethod for the diagnosis and drug susceptibility testing of M. tuberculosis. It may be a useful tool for establishing tuberculosis laboratories in resource-limited settings because it does not require expensive equipment and a high level of training. Our study may help in choosing the appropriate treatment and control of tuberculosis.