scholarly journals Rapid direct detection of pathogens for diagnosis of joint infections by MALDI-TOF MS after liquid enrichment in the BacT/Alert blood culture system

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243790
Author(s):  
Christine Noll ◽  
Azadda Nasruddin-Yekta ◽  
Pia Sternisek ◽  
Michael Weig ◽  
Uwe Groß ◽  
...  

Pathogen identification is a critical step during diagnosis of infectious diseases. Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF-MS) has become the gold standard for identification of microorganisms cultured on solid media in microbiology laboratories. Direct identification of microbes from liquid specimen, circumventing the need for the additional overnight cultivation step, has been successfully established for blood culture, urine and liquor. Here, we evaluate the ability of MALDI-TOF MS for direct identification of pathogens in synovial fluid after liquid enrichment in BacT/Alert blood culture bottles. Influence of synovial specimen quality on direct species identification with the MALDI BioTyper/Sepsityper was tested with samples inoculated from pretested native synovia with concomitant inoculation of blood or pus, or highly viscous fluid. Here, we achieved >90% concordance with culture on solid medium, and only mixed-species samples posed significant problems. Performance in routine diagnostics was tested prospectively on bottles inoculated by treating physicians on ward. There, we achieved >70% concordance with culture on solid media. The major contributors to test failure were the absence of a measurable mass signal and mixed-specimen samples. The Sepsityper workflow worked well on samples derived from BacT/Alert blood culture bottles inoculated with synovial fluid, giving concordant results to identification from solid media. Host remnant material in the inoculum, such as blood or pus, had no detrimental effect on identification score values of the BioTyper system after processing with the Sepsityper workflow, and neither had the initial viscosity of the synovial sample.

2013 ◽  
Vol 24 (4) ◽  
pp. 191-194 ◽  
Author(s):  
Manal Tadros ◽  
Astrid Petrich

Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) can be used to identify bacteria directly from positive blood and sterile fluid cultures. The authors evaluated a commercially available kit – the Sepsityper Kit (Bruker Daltonik, Germany) – and MALDI-TOF MS for the rapid identification of organisms from 80 flagged positive blood culture broths, of which 73 (91.2%) were blood culture specimens and seven (8.7%) were cerebrospinal fluid specimens, in comparison with conventional identification methods. Correct identification to the genus and species levels was obtained in 75 of 80 (93.8%) and 39 of 50 (78%) blood culture broths, respectively. Applying the blood culture analysis module, a newly developed software tool, improved the species identification of Gram-negative organisms from 94.7% to 100% and of Gram-positive organisms from 66.7% to 70%.MALDI-TOF MS is a promising tool for the direct identification of organisms cultured from sterile sites.


2021 ◽  
Author(s):  
kwenrich not provided

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can accurately identify bloodstream pathogens directly from positive blood culture bottles without the need to wait for agar plate growth. In this study, 2% sodium dodecyl sulfate (SDS) detergent was assessed to determine its benefit in the removal of interfering cellular components for testing on the Bruker Microflex LT MALDI-TOF MS instrument with the Biotyper® CA system. Additionally, the use of a heat-drying step was evaluated for performance improvement over conventional air-drying of samples on the MALDI steel target plate. The modified method with 2% SDS outperformed the in-house protocol in overall success with percentage scores of 91% and 55% ( respectively). The data results support the potential of applying a simple lysing step to an existing in-house extraction method and the use of modified drying methods. The modified techniques evaluated in this study proved beneficial for identifying most blood culture pathogens encountered in the clinical laboratory, and they can allow for reduced turnaround times and more appropriate antibiotic treatments.


2021 ◽  
Vol 70 (2) ◽  
pp. 207-213
Author(s):  
DERU LEI ◽  
PEIYING CHEN ◽  
XUETING CHEN ◽  
YUJIE ZONG ◽  
XIANGYANG LI

Rapid identification of microorganisms in urine is essential for patients with urinary tract infections (UTIs). Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been proposed as a method for the direct identification of urinary pathogens. Our purpose was to compare centrifugation-based MALDI-TOF MS and short-term culture combined with MALDI-TOF MS for the direct identification of pathogens in urine specimens. We collected 965 urine specimens from patients with suspected UTIs, 211/965 isolates were identified as positive by conventional urine culture. Compared with the conventional method, the results of centrifugation-based MALDI-TOF MS were consistent in 159/211 cases (75.4%), of which 135/159 (84.9%) had scores ≥ 2.00; 182/211 cases (86.3%) were detected using short-term culture combined with MALDI-TOF MS, of which 153/182 (84.1%) had scores ≥ 2.00. There were no apparent differences among the three methods (p = 0.135). MALDI-TOF MS appears to accelerate the microbial identification speed in urine and saves at least 24 to 48 hours compared with the routine urine culture. Centrifugation-based MALDI-TOF MS is characterized by faster identification speed; however, it is substantially affected by the number of bacterial colonies. In contrast, short-term culture combined with MALDI-TOF MS has a higher detection rate but a relatively slow identification speed. Combining these characteristics, the two methods may be effective and reliable alternatives to traditional urine culture.


Author(s):  
Hazan Zengin Canalp ◽  
Banu Bayraktar

Using MALDI-TOF MS directly from blood culture bottles reduces the time required for pathogen identification, and the turnaround times for final identification have been compared with overnight incubation from solid media in previous studies. However, identification from a short incubation of agar plates has been increasingly accepted and successfully implemented in routine laboratories, but there is no data comparing direct MALDI-TOF MS with the short-term, incubated agar plates.


2015 ◽  
Vol 64 (11) ◽  
pp. 1346-1352 ◽  
Author(s):  
Osman Altun ◽  
Silvia Botero-Kleiven ◽  
Sarah Carlsson ◽  
Måns Ullberg ◽  
Volkan Özenci

Author(s):  
Yufeng Dai ◽  
Xinyi Xu ◽  
Xue Yan ◽  
Daming Li ◽  
Wei Cao ◽  
...  

Early and rapid identification of microorganisms is critical for reducing the mortality rate caused by bloodstream infections (BSIs). The accuracy and feasibility of directly identifying pathogens in positive blood cultures by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been intensely confirmed. In this study, we combined density centrifugation and extra chemical lysis-extraction to develop an optimized method in the blood culture process, which significantly improved the effectiveness of direct identification by MALDI-TOF MS. The accuracy was evaluated by 2,032 positive blood culture samples (115 species of microorganism). The overall MALDI-TOF MS based identification rate with scores ≥ 1.700 was 87.60%. 94.06% of gram-negative bacteria were identified consistently to the genus level, followed by anaerobes (93.33%), gram-positive bacteria (84.46%), and fungi (60.87%). This protocol could obtain results within 10–20 min at a cost of less than $0.1 per sample, which saved up to 24 h in identifying 87.60% of the microorganism from positive blood cultures. This rapid and simplified protocol facilitates the direct identification of microorganism in positive blood cultures, and exhibits the advantages of cost-effective, time-saving, and easy-to-use. It could provide the causative organism of the patient to clinicians in time for targeted treatment and reduce mortality.


Pathology ◽  
2018 ◽  
Vol 50 (6) ◽  
pp. 676-679 ◽  
Author(s):  
Christopher J. McIver ◽  
Noel Er ◽  
Chinmoy Mukerjee ◽  
Sophie Tokis ◽  
Peter Taylor

2020 ◽  
Vol 154 (Supplement_1) ◽  
pp. S126-S126
Author(s):  
H E Berg ◽  
S Shannon ◽  
A N Schuetz

Abstract Introduction/Objective Matrix-assisted laser desorption ionization/ time-of-flight mass spectrometry (MALDI-TOF MS) direct from positive blood culture bottles has facilitated drastic drops in turn-around times for microorganism identification but has been poorly studied for anaerobes. We investigated the ability of MALDI-TOF to provide early anaerobe identification at 4 hours and 18-24 hours of growth on agar from anaerobic blood culture bottles. Additionally, we reviewed medical records of such patients to ascertain impact of early identification on antimicrobial treatment. Methods Over 9 months, we ran MALDI-TOF on early growth from blood cultures positive for growth in BACTEC™ Lytic/ 10 Anaerobic/F bottles. Broth from each bottle was subbed to sheep blood agar (4 hours, 5% CO2) and 2 CDC (BD-BBL™) anaerobic blood agar plates (examined 18-24 hours and 48 hours). Bruker Biotyper® RUO v7854 and Mayo Clinic Custom MALDI-TOF MS libraries were used for identification. Results 144/184 (78%) bottles resulted in growth of aerobic bacteria. Of the remaining bottles with growth of anaerobes, 38 were assessed by early MALDI-TOF. Early MALDI-TOF at 4 hours identified 3 Clostridium perfringens (8%) and an additional 26/38 (68%) isolates at 18-24 hours (both Gram-positive and –negative). Routine 48 hour identification was required for 9 (24%) isolates. In 7 cases, early MALDI-TOF resulted in a change to more appropriate antimicrobial therapy, most often for Bacteroides. Conclusion 29/38 (76%) of anaerobes from blood culture bottles were identified by early MALDI-TOF and reported to the clinician at least 24 hours before routine review of anaerobic sub plates for growth. All C. perfringens and Bacteroides were identified by 4 and 24 hours, respectively. Although early MALDI-TOF resulted in antimicrobial therapy adjustments in a minority of cases, it may allow for more targeted and earlier antimicrobial therapy. Early MALDI-TOF from anaerobic blood culture bottles should be considered for improved patient care and antimicrobial stewardship.


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