Study of aerobic and anaerobic bacterial profile of nosocomial infections and their antibiotic resistance in a referral center, Southwest Iran: A three year cross-sectional study.

BACKGROUND: The drug resistance is expected to be the most important challenge in infection control in Iran, where there is no local report or standard drug resistance monitoring system. Therefore, this study aimed to investigate the aerobic and anaerobic bacterial profile of nosocomial infections and their antibiotic resistance in Ahvaz, southwest Iran. METHODOLOGY: The gram-positive and gram-negative bacteria were identified on the basis of conventional culture and biochemical tests. The antibiotic resistance of the bacterial isolates against antibiotics was determined by the disk diffusion method. RESULTS: Among total 1156 collected positive samples, E. coli and coagulase-negative staphylococci (CoNS) were the most frequent pathogenic gram negative bacteria (GNB) and gram positive bacteria (GPB) respectively. Drug susceptibility testing revealed that among GNB, P. aeruginosa was 100% resistant to amikacin, cefepime, ciprofloxacin and tetracycline. In the case of E. coli, the resistance rate was (98%) for trimethoprim sulfamethoxazole and cefepime. For GPB, S. aureus showed the highest resistance rates to amikacin (100%) and clindamycin (100%). In addition, CoNS strains showed a high level of resistance to doxycycline (100%), erythromycin (100%) and cefoxitin (97%). In Bacteroeides fragilis isolates, the highest resistance rate belonged to clindamycin (72%), and Clostridium perfringens strains showed high level of resistance to penicillin (46%). CONCLUSION: The results highlighted that there are distinct factors leading to antimicrobial resistance in Ahvaz, southwest Iran. The primary contributors to the resistance development, include poor surveillance of drug-resistant infections, poor quality of available antibiotics, clinical misuse, and the ease of access to antibiotics. Moreover, similar factors such as self-medication and the lack of regulation on medication imports play a role in antibiotic resistance in the region.

Characterization of the most common embCAB gene mutations associated with ethambutol resistance in Mycobacterium tuberculosis isolates from Iran.

INTRODUCTION: Ethambutol (Emb) is one of the first-line drugs in the standard combination therapy for tuberculosis; however, due to the rapid increase in Emb resistance among clinical isolates of Mycobacterium tuberculosis (MTB), early detection of Emb resistance is desirable. As the embCAB operon is considered involved in resistance to Emb, this study aimed to analyze the most common mutations within the embCAB operon among MTB isolates from Iran to find any correlations of these mutations with Emb resistance. METHODS: A total of 307 clinical isolates of MTB were screened for Emb resistance by phenotypic drug-susceptibility testing. PCR amplification was performed on extracted DNA from all Emb-resistant and randomly selected Emb-susceptible isolates using sets of primers for various gene loci of embC, embA, and embB, followed by sequencing for the detection of most common alterations. RESULTS: In total, ten isolates showed resistance to Emb by phenotypic susceptibility testing (3.25%). The mutation rate in ten Emb-resistant MTB strains was 20% (n=2), comprising one mutation in embB (10%), at codon 306 Met-Val and one in embC (10%) at codon 270 Thr-Ile. A nonsynonymous mutation in the embA gene in one of the randomly selected Emb-susceptible isolates located in codon 330 Leu-Leu was also noticed. CONCLUSION: The majority of our Emb-resistant isolates (n=8, 80%) did not demonstrate the sequences investigated within the embCAB operon. As such, these mutations solely are insufficient for the development of complete resistance to Emb in MTB isolates. Additional mechanisms of resistance other than mutations in these sequences studied within the embCAB operon should also be considered.

Molecular detection of rifampin, isoniazid, and ofloxacin resistance in Iranian isolates of Mycobacterium tuberculosis by high-resolution melting analysis.

BACKGROUND: The emergence of drug resistance among Mycobacterium tuberculosis (MTB) strains is a serious health concern worldwide. The development of rapid molecular diagnostic methods in recent years has a significant impact on the early detection of resistance to major anti-TB drugs in MTB isolates, which helps in employing appropriate treatment regimen and prevents the spread of drug-resistant strains. This study was designed to evaluate the efficacy of real-time PCR and high-resolution melting (HRM) curve analysis for the determination of resistance to rifampin (RIF), isoniazid (INH), and ofloxacin (OFX) in MTB isolates and to investigate their resistance-related mutations. METHODS: HRM analysis was performed to screen 52 (32 drug-resistant and 20 fully susceptible) MTB clinical isolates for mutations in rpoB, katG, mab-inhA, and gyrA genes. The HRM results were then confirmed by DNA sequencing. RESULTS: In total, 32 phenotypically resistant isolates, comprising 18 RIF-, 16 INH-, and five OFX- resistant strains, were investigated. HRM analysis successfully identified 15 out of 18 mutations in rpoB, 14 out of 16 mutations in katG and mab-inhA, and four out of five mutations in gyrA conferring resistance to RIF, INH, and OFX, respectively. The obtained sensitivity and specificity, respectively, for HRM in comparison with phenotypic susceptibility testing were found to be 83.3% and 100% for RIF, 87.5% and 100% for INH, and 80% and 100% for OFX. In five resistant strains (12.8%), no mutation was detected by using HRM and DNA sequencing. CONCLUSION: HRM assay is a rapid, accurate, and cost-effective method possessing high sensitivity and specificity for the determination of antibiotic resistance among MTB clinical isolates and screening of their associated mutations. This method can generate results in a shorter period of time than taken by the phenotypic susceptibility testing and also allows for timely treatment and prevention of the emergence of possible MDR strains.