Journal Article

Diagnostic methods and protocols for rapid determination of methicillin resistance in Staphylococcus aureus bloodstream infections: a comparative analysis

Boattini, Matteo, Guarrasi, Luisa, Ricciardelli, Guido, Comini, Sara, Casale, Roberto, Costa, Cristina, Cavallo, Rossana, Bianco, Gabriele

2025

Overview

Purpose To evaluate diagnostic performance of four diagnostic methods for rapid determination of methicillin resistance in S. aureus positive blood cultures (BCs). Methods Clinical and spiked BCs were subjected to the evaluation of the following methods and protocols: a. Eazyplex ® MRSA Plus loop‐mediated isothermal amplification (LAMP) assay directly from BC fluid; b. MALDI-TOF MS subtyping on BC pellet extracted with Rapid Sepsityper ® protocol and on 4-h short-term subculture; c. Clearview™ Culture Colony PBP2a SA immunochromatography assay on BC pellet and on 4-h short-term subculture; d. EUCAST RAST cefoxitin screen test performed directly from BC and including reading times at 4-h, 6-h and 16–20-h. Results Eazyplex ® MRSA plus exhibited the best performance, showing 100% sensitivity, specificity, positive predictive value, and negative predictive value, followed by PBP2a SA Culture Colony Clearview assay and EUCAST RAST cefoxitin screen. MALDI-TOF MS subtyping showed the lowest diagnostic accuracy (59.8 and 65.7% directly from BC and from 4-h subculture, respectively). In detail, sensitivity and specificity ranged from 24.3% to 20.4% and from 88.9% to 98.3% for protocols performed from BC pellet and 4-h subculture, respectively. Conclusions The Eazyplex ® MRSA Plus and the immunochromatographic Clearview™ PBP2a SA Culture Colony methods can provide reliable results within 1 h from the start of positive BC processing. MALDI TOF MS subtyping showed unacceptable specificity by performing analysis from BC pellets, while its sensitivity depends on the prevalence of PSM-positive MRSA strains. The EUCAST RAST, based on disc diffusion, showed excellent performance with a time-to-result of at least 4 h.

• Publisher: Springer Berlin Heidelberg,Springer Nature B.V
• Subject: Anti-Bacterial Agents - pharmacology / Antibiotic resistance / Assaying / Bacteremia - diagnosis / Bacteremia - microbiology / Biomedical and Life Sciences / Biomedicine / Blood Culture / Cefoxitin / Comparative analysis / Drug resistance / Humans / Internal Medicine / Medical Microbiology / Methicillin / Methicillin Resistance / Methicillin-Resistant Staphylococcus aureus - drug effects / Methicillin-Resistant Staphylococcus aureus - genetics / Methicillin-Resistant Staphylococcus aureus - isolation & purification / Microbial Sensitivity Tests / Molecular Diagnostic Techniques - methods / Nucleic Acid Amplification Techniques - methods / Pellets / Performance evaluation / Sensitivity and Specificity / Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Staphylococcal Infections - diagnosis / Staphylococcal Infections - microbiology / Staphylococcus infections / Subculture