Microorganisms direct identification from blood culture by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) allows a fast and reliable bacterial identification from culture plates. Direct analysis of clinical samples may increase its usefulness in samples in which a fast identification of microorganisms can guide empirical treatment, such as blood cultures (BC). Three hundred and thirty BC, reported as positive by the automated BC incubation device, were processed by conventional methods for BC processing, and by a fast method based on direct MALDI-TOF MS. Three hundred and eighteen of them yield growth on culture plates, and 12 were false positive. The MALDI-TOF MS-based method reported that no peaks were found, or the absence of a reliable identification profile, in all these false positive BC. No mixed cultures were found. Among these 318 BC, we isolated 61 Gram-negatives (GN), 239 Gram-positives (GP) and 18 fungi. Microorganism identifications in GN were coincident with conventional identification, at the species level, in 83.3% of BC and, at the genus level, in 96.6%. In GP, identifications were coincident with conventional identification in 31.8% of BC at the species level, and in 64.8% at the genus level. Fungaemia was not reliably detected by MALDI-TOF. In 18 BC positive for Candida species (eight C. albicans, nine C. parapsilosis and one C. tropicalis), no microorganisms were identified at the species level, and only one (5.6%) was detected at the genus level. The results of the present study show that this fast, MALDI-TOF MS-based method allows bacterial identification directly from presumptively positive BC in a short time (<30 min), with a high accuracy, especially when GN bacteria are involved.

Ferreira L ，Sánchez-Juanes F ，Porras-Guerra I ，García-García MI ，García-Sánchez JE ，González-Buitrago JM ，Muñoz-Bellido JL ... -  《-》

Identification of pathogenic microorganisms directly from positive blood vials by matrix-assisted laser desorption/ionization time of flight mass spectrometry.

Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is a promising and fast method for identifying fungi and bacteria directly from positive blood cultures. Various pre-treatment methods for MALDI-TOF MS identification have been reported for this purpose. In-house results for identification of bacterial colonies by MALDI-TOF MS using a cut-off score of 1.5 did not reduce the diagnostic accuracy compared with the recommended cut-off score of 1.8. A 3-month consecutive study of positive blood cultures was carried out in our laboratory to evaluate whether the Sepsityper™ Kit (Bruker Daltonics) with Biotyper 2.0 software could be used as a fast diagnostic tool for bacteria and fungi and whether a 1.5 cut-off score could improve species identification compared with the recommended score of 1.8. Two hundred and fifty-six positive blood vials from 210 patients and 19 blood vials spiked with fungi were examined. Using the cut-off score of 1.8, 81% Gram-negative bacteria were identified to the species level compared to 84% using a cut-off score of 1.5. For Gram-positive bacteria 44% were identified to the species level with a cut-off of 1.8 compared to 55% with the value of 1.5. The overall identification rate was 63% (cut-off 1.5) and 54% (cut-off 1.8). Seventy-seven per cent of fungal species were identified with both log scores. MALDI-TOF MS was in this study found to be a powerful tool in fast diagnosis of Gram-negative bacteria and fungi and to a lesser degree of Gram positives. Using 1.5 as cut-off score increased the diagnosis for both Gram-positives and -negatives bacteria.

Nonnemann B ，Tvede M ，Bjarnsholt T 《-》

Real-time identification of bacteria and Candida species in positive blood culture broths by matrix-assisted laser desorption ionization-time of flight mass spectrometry.

Ferroni A ，Suarez S ，Beretti JL ，Dauphin B ，Bille E ，Meyer J ，Bougnoux ME ，Alanio A ，Berche P ，Nassif X ... -  《-》

Detection of microorganisms in blood specimens using matrix-assisted laser desorption ionization time-of-flight mass spectrometry: a review.

Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) initiated a revolution in the identification of organisms grown on solid medium, including bacteria and fungi. Rapid identification of organisms responsible for septicaemia, which are typically grown in broth, is now expanding the field of application. Despite the fact that there are fewer than ten reports in the literature, published data indicate that MALDI-TOF MS yields accurate identification of blood-borne organisms in ≥80% of cases for inocula of >10⁷ organisms/mL. A major current limitation is failure to accurately identify Streptococcus pneumoniae among viridans steptococci. Identification is achieved in <2 h, sharply reducing the turn-around time for communication of identification to the clinician. Further progress in handling protocols and automation, and extraction of antibiotic resistance data from the MALDI-TOF MS spectra, will further push this emerging approach as the standard one in the laboratory diagnosis of septicaemia, paving the way to application in further clinical situations and clinical specimens.

Drancourt M 《-》

The sensitivity of direct identification from positive BacT/ALERT™ (bioMérieux) blood culture bottles by matrix-assisted laser desorption ionization time-of-flight mass spectrometry is low.

Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been presented as a novel method for the direct identification of bacteria from positive blood culture bottles. The rate of the MALDI TOF MS-based identification in the present study from positive BacT/ALERT (bioMérieux, Marcy l'Etoile, France) blood culture bottles was 30%, which is far below the previously reported sensitivities using the BACTEC (Becton Dickinson, Franklin Lakes, NJ, USA) system. We also found evidence that the Biotyper algorithm did not identify a second pathogen in cases of positive BacT/ALERT blood culture bottles containing two different species.

Szabados F ，Michels M ，Kaase M ，Gatermann S ... -  《-》