A rapid diagnostic workflow for cefotaxime-resistant Escherichia coli and Klebsiella pneumoniae detection from blood cultures by MALDI-TOF mass spectrometry.

BACKGROUND: Nowadays, the global spread of resistance to oxyimino-cephalosporins in Enterobacteriaceae implies the need for novel diagnostics that can rapidly target resistant organisms from these bacterial species. METHODS: In this study, we developed and evaluated a Direct Mass Spectrometry assay for Beta-Lactamase (D-MSBL) that allows direct identification of (oxyimino)cephalosporin-resistant Escherichia coli or Klebsiella pneumoniae from positive blood cultures (BCs), by using the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) technology. RESULTS: The D-MSBL assay was performed on 93 E. coli or K. pneumoniae growing BC samples that were shortly co-incubated with cefotaxime (CTX) as the indicator cephalosporin. Susceptibility and resistance defining peaks from the samples' mass spectra were analyzed by a novel algorithm for bacterial organism classification. The D-MSBL assay allowed discrimination between E. coli and K. pneumoniae that were resistant or susceptible to CTX with a sensitivity of 86.8% and a specificity of 98.2%. CONCLUSION: The proposed algorithm-based D-MSBL assay, if integrated in the routine laboratory diagnostic workflow, may be useful to enhance the establishment of appropriate antibiotic therapy and to control the threat of oxyimino-cephalosporin resistance in hospital.

Comparison of Mycoplasma IES, Mycofast Revolution and Mycoplasma IST2 to detect genital mycoplasmas in clinical samples.

INTRODUCTION: Culture is regarded as the gold standard for the detection of genital mycoplasma in clinical samples. Commercially available diagnostic kits, based on liquid broth cultures, provide interesting alternatives to conventional culture. We assessed the laboratory performances of Mycoplasma IES (IES), the Mycofast Revolution (REV) and Mycoplasma IST 2 (IST2) compared to A7 agar plates for the detection of Ureaplasma urealyticum and Mycoplasma hominis in clinical samples. METHODOLOGY: From April to July 2013, endocervical or vaginal samples were collected from sexually active women with abnormal vaginal discharge. Each specimen was tested in parallel using the three commercial kits and the A7 agar plates. RESULTS: A total of 303 samples were included in this study, 35.6% (108/303) of which were positive on A7 plates. Sensitivities for the detection of U. urealyticum of IES, REV and IST2 were 100%, 96.2% and 95.3%, respectively while those for M. hominis were of 92.8%, 92.8% and 85.7%, respectively. Specificity was 100% for the 3 methods. Concerning antimicrobial susceptibility testing, full agreement between IES and REV was documented. CONCLUSIONS: The Mycoplasma IES kit was found to be equivalent or superior compared to other commercial culture-based assays for a rapid and accurate identification of U. urealyticum and M. hominis and detection of resistance. It might be considered a cost-effective tool for detection of these organisms, particularly attractive in developing countries.