Distribution of serotypes and antibiotic resistance of invasive Pseudomonas aeruginosa in a multi-country collection

Background Pseudomonas aeruginosa is an opportunistic pathogen that causes a wide range of acute and chronic infections and is frequently associated with healthcare-associated infections. Because of its ability to rapidly acquire resistance to antibiotics, P. aeruginosa infections are difficult to treat. Alternative strategies, such as a vaccine, are needed to prevent infections. We collected a total of 413 P. aeruginosa isolates from the blood and cerebrospinal fluid of patients from 10 countries located on 4 continents during 2005–2017 and characterized these isolates to inform vaccine development efforts. We determined the diversity and distribution of O antigen and flagellin types and antibiotic susceptibility of the invasive P. aeruginosa. We used an antibody-based agglutination assay and PCR for O antigen typing and PCR for flagellin typing. We determined antibiotic susceptibility using the Kirby-Bauer disk diffusion method. Results Of the 413 isolates, 314 (95%) were typed by an antibody-based agglutination assay or PCR (n = 99). Among the 20 serotypes of P. aeruginosa, the most common serotypes were O1, O2, O3, O4, O5, O6, O8, O9, O10 and O11; a vaccine that targets these 10 serotypes would confer protection against more than 80% of invasive P. aeruginosa infections. The most common flagellin type among 386 isolates was FlaB (41%). Resistance to aztreonam (56%) was most common, followed by levofloxacin (42%). We also found that 22% of strains were non-susceptible to meropenem and piperacillin-tazobactam. Ninety-nine (27%) of our collected isolates were resistant to multiple antibiotics. Isolates with FlaA2 flagellin were more commonly multidrug resistant (p = 0.04). Conclusions Vaccines targeting common O antigens and two flagellin antigens, FlaB and FlaA2, would offer an excellent strategy to prevent P. aeruginosa invasive infections.

A New Colorimetric Method for Rapid Detection of Antibiotic Resistance in Escherichia coli Isolates

Background: The quick diagnosis and early initiation of antibiotic therapy in bacteria-induced infections is of paramount importance. Accordingly, the rapid identification of the causative agent, the short-term results of antibiotic sensitivity, the selection and use of right antibiotics for treatment further highlights the significance of this issue. Objectives: This study aimed to develop a new susceptibility testing method to provide rapid results in Escherichia coli clinical isolates and report the antibiotic susceptibility test results to clinicians in a short period. Methods: In the study, one hundred and ten E. coli clinical isolates were tested. In this regard, antibiotics recommended by the "Clinical and Laboratory Standards Institute (CLSI)" for testing the sensitivity of E. coli isolates, including amoxicillin-clavulanate, cefixime, ceftriaxone, ertapenem, ciprofloxacin, gentamicin, trimethoprim-sulfamethoxazole, and nitrofurantoin were tested. For quality control, E. coli ATCC25922, E. coli ATCC35218, Staphylococcus aureus ATCC29213, and E. coli 13846NTCC strains were used. The broth microdilution method recommended by CLSI was used as the reference method. Minimum inhibitory concentration values were determined, and antimicrobial susceptibilities were then determined according to the “European Committee on Antimicrobial Susceptibility Testing (EUCAST)” criteria. In the next phase, the results of the resazurin microplate method (RMM) were compared. Results: The comparison of the RMM developed in the present study with the reference method revealed that the calculated essential agreement ratios for eight antibiotics varied from 82.72 to 100%, and the categorical agreement values ranged from 95.45 to100%. Conclusions: According to the findings, the RMM results were highly in agreement with the results of the reference method. RMM allows the detection of antibiotic susceptibility quickly (e.g., within 5 hours) as such it is preferred, especially for laboratories with limited facilities. However, further multi-center studies are recommended to use this method in routine laboratories.