scholarly journals Evaluation of Rapid Sepsityper® protocol and specific MBT-Sepsityper module (Bruker Daltonics) for the rapid diagnosis of bacteremia and fungemia by MALDI-TOF-MS

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Léa Ponderand ◽  
Patricia Pavese ◽  
Danièle Maubon ◽  
Emmanuelle Giraudon ◽  
Thomas Girard ◽  
...  
Keyword(s):  
Formic Acid ◽  
Bloodstream Infections ◽  
Rapid Identification ◽  
Rapid Diagnosis ◽  
Blood Cultures ◽  
Acid Extraction ◽  
Gram Negative Bacteria ◽  
Reliable Identification ◽  
Extraction Step ◽  
Bruker Daltonics

AbstractDuring bloodstream infections, rapid adaptation of empirical treatment according to the microorganism identified is essential to decrease mortality. The aim of the present study was to assess the microbiological performances of a new rapid version of the Sepsityper® kit (Bruker Daltonics) allowing identification of bacteria and yeast by MALDI-TOF mass spectrometry directly from positive blood cultures in 10 min and of the specific MBT-Sepsityper module for spectra analysis, designed to increase identification performance. Identification rates were determined prospectively on 350 bacterial and 29 fungal positive blood cultures, and compared to conventional diagnostic method. Our rapid diagnosis strategy (Rapid Sepsityper® protocol: one spot with and one without formic acid extraction step) combined to MBT-Sepsityper module provided 65.4%, 78.9% and 62% reliable identification to the species level of monomicrobial positive blood cultures growing respectively Gram-positive, Gram-negative bacteria or yeast. Importantly, identification rates of Gram-positive bacteria were higher in anaerobic than in aerobic bottles (77.8% vs 22.2%; p = 0.004), if formic acid extraction step was performed (60.8% vs 39.2%; p = 1.8e−6) and if specific MBT-Sepsityper module was used (76.2% vs 61.9%, p = 0.041) while no significant differences were observed for Gram-negative bacteria. For yeasts identification, formic acid extraction step improved rapid identification rate by 37.9% while the specific MBT-Sepsityper module increased overall performances by 38%, providing up to 89.7% reliable identification if associated with the standard Sepsityper® protocol. These performances, associated with a reduce turnaround time, may help to implement a rapid identification strategy of bloodstream infections in the routine workflow of microbiology laboratories.

scholarly journals Evaluation of Rapid Immunochromatographic Tests for the Direct Detection of Extended Spectrum Beta-Lactamases and Carbapenemases in Enterobacterales Isolated from Positive Blood Cultures

2021 ◽  
Author(s):  
Ahmed S. Keshta ◽  
Nazik Elamin ◽  
Mohammad Rubayet Hasan ◽  
Andrés Pérez-López ◽  
Diane Roscoe ◽  
...  
Keyword(s):  
Direct Detection ◽  
Bloodstream Infections ◽  
Resistance Mechanisms ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Gram Negative Bacteria ◽  
Beta Lactamases ◽  
Extended Spectrum Beta Lactamase ◽  
Extended Spectrum ◽  
Extended Spectrum Beta Lactamases

The incidence of bloodstream infections (BSI) with extended spectrum beta-lactamase (ESBL) producing and carbapenemase producing Enterobacterales (CPE) is increasing at an alarming rate, for which only limited therapeutic options remain available. Rapid identification of these bacteria along with their antibiotic resistance mechanisms in positive blood cultures with Gram-negative bacteria will allow for early initiation of effective therapy and limit the overuse of broad-spectrum antibiotics in BSI (1).

scholarly journals Trends of Bloodstream Infections in a University Greek Hospital during a Three-Year Period: Incidence of Multidrug-Resistant Bacteria and Seasonality in Gram-negative Predominance

2017 ◽  
Vol 66 (2) ◽  
pp. 171-180 ◽  
Author(s):  
Fevronia Kolonitsiou ◽  
Matthaios Papadimitriou-Olivgeris ◽  
Anastasia Spiliopoulou ◽  
Vasiliki Stamouli ◽  
Vasileios Papakostas ◽  
...  
Keyword(s):  
Bloodstream Infections ◽  
Multidrug Resistant ◽  
Blood Cultures ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Gram Negative Bacteria ◽  
Multidrug Resistant Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Carbapenem Resistant

The aim of the study was to assess the epidemiology, the incidence of multidrug-resistant bacteria and bloodstream infections’ (BSIs) seasonality in a university hospital. This retrospective study was carried out in the University General Hospital of Patras, Greece, during 2011–13 y. Blood cultures from patients with clinical presentation suggestive of bloodstream infection were performed by the BacT/ALERT System. Isolates were identified by Vitek 2 Advanced Expert System. Antibiotic susceptibility testing was performed by the disk diffusion method and E-test. Resistance genes (mecA in staphylococci; vanA/vanB/vanC in enterococci; blaKPC/blaVIM/blaNDM in Klebsiella spp.) were detected by PCR. In total, 4607 (9.7%) blood cultures were positive from 47451 sets sent to Department of Microbiology, representing 1732 BSIs. Gram-negative bacteria (52.3%) were the most commonly isolated, followed by Gram-positive (39.5%), fungi (6.6%) and anaerobes bacteria (1.8%). The highest contamination rate was observed among Gram-positive bacteria (42.3%). Among 330 CNS and 150 Staphylococcus aureus, 281 (85.2%) and 60 (40.0%) were mecA-positive, respectively. From 113 enterococci, eight were vanA, two vanB and two vanC-positives. Of the total 207 carbapenem-resistant Klebsiella pneumoniae (73.4%), 202 carried blaKPC, four blaKPC and blaVIM and one blaVIM. A significant increase in monthly BSIs’ incidence was shown (R2: 0.449), which may be attributed to a rise of Gram-positive BSIs (R2: 0.337). Gram-positive BSIs were less frequent in spring (P < 0.001), summer (P < 0.001), and autumn (P < 0.001), as compared to winter months, while Gram-negative bacteria (P < 0.001) and fungi (P < 0.001) were more frequent in summer months. BSIs due to methicillin resistant S. aureus and carbapenem-resistant Gram-negative bacteria increased during the study period. The increasing incidence of BSIs can be attributed to an increase of Gram-positive BSI incidence, even though Gram-negative bacteria remained the predominant ones. Seasonality may play a role in the predominance of Gram-negative’s BSI.

scholarly journals Microbiological characteristics of bloodstream infections in a reference hospital in northeastern Brazil

2024 ◽  
Vol 84 ◽  
Author(s):  
M. C. Melo ◽  
A. P. M. Carvalho Neto ◽  
T. L. G. Q. Maranhão ◽  
E. S. Costa ◽  
C. M. A. Nascimento ◽  
...  
Keyword(s):  
Public Hospital ◽  
Anaerobic Bacteria ◽  
Bloodstream Infections ◽  
Blood Cultures ◽  
Northeastern Brazil ◽  
Gram Negative Bacteria ◽  
Gram Positive Bacteria ◽  
Microbiological Characteristics ◽  
Reference Hospital ◽  
Aerobic And Anaerobic

Abstract Routine blood culture is used for the detection of bloodstream infections by aerobic and anaerobic bacteria and by common pathogenic yeasts. A retrospective study was conducted in a public hospital in Maceió-AL, by collecting data of all medical records with positive blood cultures. Out of the 2,107 blood cultures performed, 17% were positive with Staphylococcus coagulase negative (51.14%), followed by Staphylococcus aureus (11.21%) and Klebsiella pneumoniae (6.32%). Gram-positive bacteria predominated among positive blood cultures, highlighting the group of Staphylococcus coagulase-negative. While Gram-negative bacteria had a higher number of species among positive blood cultures.

Optimization of MALDI-ToF mass spectrometry for yeast identification: a multicenter study

2019 ◽  
Vol 58 (5) ◽  
pp. 639-649 ◽  
Author(s):  
Anne-Cécile Normand ◽  
Frédéric Gabriel ◽  
Arnaud Riat ◽  
Carole Cassagne ◽  
Nathalie Bourgeois ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Formic Acid ◽  
Extraction Method ◽  
Acid Extraction ◽  
Identification Performance ◽  
Yeast Identification ◽  
Maldi Tof ◽  
Extraction Step ◽  
The Impact ◽  
Target Database

Abstract Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) is routinely used in mycology laboratories to rapidly identify pathogenic yeasts. Various methods have been proposed to perform routine MS-based identification of clinically relevant species. In this study, we focused on Bruker technology and assessed the identification performance of three protocols: two pretreatment methods (rapid formic acid extraction directly performed on targets and full extraction using formic acid/acetonitrile in tubes) and a direct deposit protocol that omits the extraction step. We also examined identification performance using three target types (ground-steel, polished-steel, and biotargets) and two databases (Bruker and online MSI [biological-mass-spectrometry-identification application]) in a multicenter manner. Ten European centers participated in the study, in which a total of 1511 yeast isolates were analyzed. The 10 centers prospectively performed the three protocols on approximately 150 yeast isolates each, and the corresponding spectra were then assessed against two reference spectra databases (MSI and Bruker), with appropriate thresholds. Three centers evaluated the impact of the targets. Scores were compared between the various combinations, and identification accuracy was assessed. The protocol omitting the extraction step was inappropriate for yeast identification, while the full extraction method yielded far better results. Rapid formic acid extraction yielded variable results depending on the target, database and threshold. Selecting the optimal extraction method in combination with the appropriate target, database and threshold may enable simple and accurate identification of clinically relevant yeast samples. Concerning the widely used polished-steel targets, the full extraction method still ensured better scores and better identification rates.

scholarly journals Evaluation of Verigene Blood Culture Test Systems for Rapid Identification of Positive Blood Cultures

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Jae-Seok Kim ◽  
Go-Eun Kang ◽  
Han-Sung Kim ◽  
Hyun Soo Kim ◽  
Wonkeun Song ◽  
...  
Keyword(s):  
Blood Culture ◽  
Resistance Genes ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Single Strain ◽  
Gram Negative ◽  
Gram Positive Bacteria ◽  
Susceptibility Tests

The performance of molecular tests using the Verigene Gram-Positive and Gram-Negative Blood Culture nucleic acid tests (BC-GP and BC-GN, resp.; Naosphere, Northbrook, IL, USA) was evaluated for the identification of microorganisms detected from blood cultures. Ninety-nine blood cultures containing Gram-positive bacteria and 150 containing Gram-negative bacteria were analyzed using the BC-GP and BC-GN assays, respectively. Blood cultures were performed using the Bactec blood culture system (BD Diagnostic Systems, Franklin Lakes, NJ, USA) and conventional identification and antibiotic-susceptibility tests were performed using a MicroScan system (Siemens, West Sacramento, CA, USA). When a single strain of bacteria was isolated from the blood culture, Verigene assays correctly identified 97.9% (94/96) of Gram-positive bacteria and 93.8% (137/146) of Gram-negative bacteria. Resistance genesmecAandvanAwere correctly detected by the BC-GP assay, while the extended-spectrumβ-lactamase CTX-M and the carbapenemase OXA resistance gene were detected from 30 cases cultures by the BC-GN assay. The BC-GP and BC-GN assays showed high agreement with conventional identification and susceptibility tests. These tests are useful for rapid identification of microorganisms and the detection of clinically important resistance genes from positive Bactec blood cultures.

Early Quantitative Determination of Serum Procalcitonin Can Help Distinguishing Strains of Different Bloodstream Infections in Patients with Hematologic Diseases

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 3703-3703
Author(s):  
Xiaofeng Luo ◽  
Jinhua Ren ◽  
Zhizhe Chen ◽  
Ting Yang ◽  
Jianda Hu
Keyword(s):  
Quantitative Determination ◽  
Bloodstream Infection ◽  
Large Population ◽  
Bloodstream Infections ◽  
Blood Cultures ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Hematologic Diseases ◽  
Fungi Infection

Abstract High procalcitonin (PCT) levels are strongly associated with systemic bacterial infections. PCT is produced in response to bacterial endotoxin and inflammatory cytokines. Few studies are available in the literature on PCT ability to distinguish different strains of bloodstream infections in patients with hematologic diseases. The aim of the present study was to explore the value of determining serum PCT values early, i.e., as soon as blood cultures are positive, in a large population of patients with hematologic diseases. Patients with hematologic diseases admitted to the hematology department of our hospitalfrom January 2013 to March 2016 who had bloodstream infections were retrospectively analyzed. Patients whose blood samples were collected for simultaneous blood culture and PCT test were enrolled in the study, and they were divided into agranulocytosis and non-agranulocytosis groups. Automatic microbial analyzer was used to identify all strains, and PCT levels were analyzed with an automatic electrochemiluminescence system. The relationship between PCT levels and the strains in bloodstream infections was analyzed and compared, and the diagnostic efficacy of PCT was evaluated using the receiver operating characteristic (ROC) curve. A total of 494 bloodstream infection cases that fulfilled the inclusion criteria were included in the study, involving 312 cases of bloodstream infection with single Gram-negative, 146 cases with single Gram-positive, 12 cases with single fungi, 19 cases with polymicrobes, and 5 cases identified as contaminated specimens. Unpaired t-test was used for data analysis. PCT levels for single Gram-negative infection (15.17±2.11 ng/ml) were significantly higher than those for Gram-positive infection (3.30 ± 0.93 ng/ml) (P<0.0001), or those for single fungi infection (0.22 ± 0.04 ng/ml) (P<0.0001). PCT levels for single Gram-positive infection were also significantly higher than those in single fungi infection (P<0.01). In the agranulocytosis group, which included 403 cases, the PCT levels in the single Gram-negative infection (14.14 ± 2.13 ng/ml) were significantly higher than those in single Gram-positive (2.49 ± 0.73 ng/ml) (P<0.0001), or in single fungi infection (0.24 ± 0.04 ng/ml) (P<0.0001). The PCT levels in the single Gram-positive bacterial infection were also significantly higher than those in single fungi infection (P<0.01). In the single Gram-negative bacteria bloodstream infection, we further found that the PCT levels in Enterobacteriaceae infection (17.00 ± 3.04 ng/ml) were significantly higher than those in nonfermentative Gram-negatives infection (6.49 ± 1.50 ng/ml) (P<0.01). ROC analysis was performed on monomicrobial blood cultures. ROC of single Gram-negative and Gram-positive infections revealed that the area under the curve (AUC) was 0.687, the best cut-off value was 0.58 ng/ml, the sensitivity was 60.81% and specificity was 71%. ROC of single Gram-negative and fungi infections revealed that the AUC was 0.795, the best cut-off value was 0.42 ng/ml, the sensitivity was 67% and specificity was 100%. ROC of single Gram-positive and fungi infections revealed that the AUC was 0.6, the best cut-off value was 0.44 ng/ml, the sensitivity was 37% and specificity was 100%. In the non-agranulocytosis group, we only found that the PCT levels in the single Gram-negative infection were significantly higher than those in single Gram-positive infection (P<0.05). In summary, early serum PCT quantitative determination can be used as a routine test to help to distinguish Gram-negative bacteria, Gram-positive bacteria, or fungi bloodstream infections in patients with hematologic diseases. These findings will be of great clinical value to select appropriate antibiotics for patients with hematologic diseases and bloodstream infections. Figure Figure. Disclosures No relevant conflicts of interest to declare.

scholarly journals 300. Pediatric Center Evaluation of the BioFire® Blood Culture Identification 2 Panel Versus the Original BioFire®FilmArray® Blood Culture Identification Panel for the Detection of Microorganisms and Resistance Markers in Positive Blood Cultures

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S148-S149
Author(s):  
Kristina B Pierce ◽  
Rebecca Barr ◽  
Aubrie Hopper ◽  
Charlotte Bowerbank ◽  
Anne Shaw ◽  
...  
Keyword(s):  
Blood Culture ◽  
False Negative ◽  
Bloodstream Infections ◽  
Turnaround Time ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Species Level ◽  
Genus Level ◽  
Antimicrobial Resistance Genes ◽  
Culture Identification

Abstract Background Studies show a rising annual incidence of severe sepsis, with bloodstream infections continuing to impact children. Rapid identification of causative agents and timely administration of targeted therapy can positively impact patient outcomes and improve antibiotic stewardship. The BioFire® Blood Culture Identification 2 (BCID2) Panel (BioFire Diagnostics, LLC), an updated version of the FDA-cleared BioFire® FilmArray® Blood Culture Identification (BCID) Panel, designed for use on positive blood cultures (PBCs), assesses 43 analytes, including 17 novel analytes (8 bacterial, 2 fungal, and 7 antimicrobial resistance genes), with a similar turnaround time. Methods De-identified residual PBCs for which clinician-ordered testing per standard of care (SoC) had been performed were enrolled and tested with an Investigation-Use-Only version of the BCID2 Panel. Only one positive bottle per patient was enrolled. Results of BCID2 and BCID were compared. Results 116 PBCs (48 aerobic and 68 anaerobic) were evaluated using the BioFire BCID2 Panel and results were compared to the BioFire BCID Panel. Of the 116 cases, 103 were positive on both the BioFire BCID2 Panel and the BioFire BCID Panel. Ten cases were negative on both tests. While the two panels showed 97% agreement, three cases were discrepant. Using culture (SoC) as the tiebreaker, two cases were false positive and one case was false negative on the BioFire BCID Panel. In all three cases, results from culture and the BioFire BCID2 Panel were in agreement. As expected, no organisms were detected on the BioFire BCID2 Panel in PBCs from 10% (12/116) of PBC bottles where culture identified only organisms that are not part of the panel menu. With the BioFire BCID2 Panel’s expanded platform, two cases identified as Enterobacteriaceae on the BioFire BCID Panel were identified to the genus level on the BioFire BCID2 Panel; 31 cases detected to the genus level on the BioFire BCID Panel were identified to the species level on the BioFire BCID2 Panel. Conclusion Overall, the BioFire BCID2 Panel performed well against the BioFire BCID Panel for identification of bloodstream pathogens and provided additional discrimination of some pathogens to the genus or species level. Data presented are from assays that have not been cleared or approved for diagnostic use. Disclosures All Authors: No reported disclosures

scholarly journals Introduction of Gram negative microrganisms rapid identification and direct susceptibility testing to reduce turnaround time in blood cultures

2017 ◽  
Vol 32 (4) ◽  
Author(s):  
Maria Vittoria Mauro ◽  
Saveria Dodaro ◽  
Daniela Perugini ◽  
Francesca Greco ◽  
Cristina Giraldi
Keyword(s):  
Antimicrobial Therapy ◽  
Susceptibility Testing ◽  
Infectious Agent ◽  
Direct Methods ◽  
Bloodstream Infections ◽  
Turnaround Time ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Successful Outcome ◽  
Microbial Identification

Background. Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. Beginning antimicrobial therapy early is vital for the treatment of bloodstream infections and sepsis. Reducing the time for microbial identification and antimicrobial susceptibility testing could decrease the average time needed for an appropriate antimicrobial therapy which leads to a decrease in mortality, a shortened hospital stay, and lower hospitalization costs.Methods. The direct identification and the antibiotic susceptibility testing evaluated with disk diffusion directly from blood cultures provided excellent results in Gram-negative.Results and Conclusions. Our study reveals the importance of direct methods that significantly reduce the turn around time and therefore has an impact on the successful outcome of patients suffering from bloodstream infections.

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