Evaluation of the EUCAST Rapid Antimicrobial Susceptibility Test for Enterobacterales-Containing Blood Cultures in China.

Bloodstream infection (BSI) is defined by the presence of microbes in the bloodstream and has high mortality. Early antimicrobial therapy is key to treating BSI patients. Because of potential antimicrobial resistance, rapid evaluation for the most suitable antimicrobial therapy is important for appropriate treatment. In China, the current workflow of microbiological diagnosis in BSI involves blood culture, species identification, and antimicrobial susceptibility testing, which takes around 3 days. However, this delay could lead to worse symptoms. To rapidly and accurately assess antimicrobial susceptibility, in this study, we applied EUCAST rapid antimicrobial susceptibility testing (RAST) to determine the antimicrobial susceptibilities of the most frequently detected Enterobacterales sampled in China, including Escherichia coli and Klebsiella pneumoniae. Based on EUCAST guidelines, we evaluated its efficiencies with six commercially available antimicrobials, including imipenem (10 µg), meropenem (10 µg), ciprofloxacin (5 µg), levofloxacin (5 µg), amikacin (30 µg), and trimethoprim-sulfamethoxazole (1.25/23.75 µg), with bacterium-spiked blood cultures. In addition, we developed potential breakpoints for a recently introduced antimicrobial, 30/20 µg ceftazidime-avibactam, which has high potential for treating multidrug-resistant Enterobacterales. Our results showed that EUCAST RAST is a reliable method for rapidly determining the antimicrobial susceptibilities of BSI-causing bacteria in China, with an overall categorical agreement rate at 8 h of ≥90%. The breakpoints developed in this study can categorize the isolates sampled in this study with an accuracy of 93%. Results from our experiments can be applied to clinically determine the microbial susceptibility of BSI-causing bacteria within 8 h and benefit clinical diagnostics for BSI patients.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for the identification of beta-hemolytic streptococci.

OBJECTIVE: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged as promising technology for species identification. The purpose of this investigation was to compare the performance of MS and the traditional method for identification of beta-hemolytic streptococci (BHS). METHODS: Clinical BHS isolates were identified by the BD Phoenix SMIC/ID Streptococcal panels, and two MALDI-TOF MS platforms: the VITEK MS and the Bruker MALDI Biotyper systems respectively. In case of discordant results, 16sRNA sequencing was performed to provide the reference ID. RESULTS: A total of 96 isolates of BHS were analyzed. Thirty-six isolates (20.8%) were re-tested by BD Phoenix for identification failure; and four isolates (4.2%) were rerun on the Bruker system for low identification score. No isolate need a second run for identification by Vitek MS system. Overall, BD Phoenix, BioTyper and Vitek MS automated system accurately identified 76 strains (79.2%), 91 (94.7%) strains and 92 (95.8%) strains, respectively. CONCLUSIONS: Our study suggests that MALDI-TOF MS is a superior method to conventional phenotypic methods for BHS identification.

The value of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in identifying clinically relevant bacteria: a comparison with automated microbiology system.

BACKGROUND: Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been developed as a new-type soft ionization mass spectrometry in the recent year. Increasing number of clinical microbiological laboratories consider it as an innovate approach for bacterial identification. METHODS: A total of 876 clinical strains, comprising 52 species in 27 genus, were obtained from Fudan University Affiliated Zhongshan Hospital. We compared the identification accuracy of the Vitek MS system (bioMerieux, Marcy l'Etoile) to other conventional methods for bacterial identification. 16S rRNA gene sequencing was performed as a reference identification method in cases of discrepant results. RESULTS: The Vitek MS system consistently produced accurate results within minutes of loading, while conventional methods required several hours to produce identification results. Among the 876 isolates, the overall performance of Vitek MS was significantly better than the conventional method both for correct species identification (830, 94.7% vs. 746, 85.2%, respectively, P=0.000). CONCLUSIONS: Compared to traditional identification methods, MALDI-TOF MS is a rapid, accurate and economical technique to enhance the clinical value of microorganism identification.