Applicability of an in-House Saponin-Based Extraction Method in Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Bacterial and Fungal Species in Positively Flagged Blood Cultures

Chien, Jung-Yien; Lee, Tai-Fen; Du, Shin-Hei; Teng, Shih-Hua; Liao, Chun-Hsing; Sheng, Wang-Hui; Teng, Lee-Jene; Hsueh, Po-Ren

doi:10.3389/fmicb.2016.01432

Cited by 19 publications

(9 citation statements)

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“…In addition, increased error rates have been documented for polymicrobial in comparison to monomicrobial BCs in the context of direct MALDI-TOF MS [24]. Only a few studies reported that it may be possible to identify multiple microbial species in polymicrobial BCs by MALDI-TOF MS [16, 19, 25, 26]; however, the correct identification of the two different microorganisms was generally limited to a relatively low percentage of polymicrobial cultures [16, 19, 26]. In a recent study, the performance of the Bruker MBT Sepsityper IVD module was evaluated for the direct identification of microorganisms in polymicrobial BCs by MALDI-TOF MS [26].…”

Section: Discussionmentioning

confidence: 99%

A new culture-based method for rapid identification of microorganisms in polymicrobial blood cultures by MALDI-TOF MS

et al. 2019

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BackgroundThe application of matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) to microbial identification has allowed the development of rapid methods for identification of microorganisms directly in positive, blood cultures (BCs). These methods can yield accurate results for monomicrobial BCs, but often fail to identify multiple microorganisms in polymicrobial BCs. The present study was aimed at establishing a rapid and simple method for identification of bacteria and yeast in polymicrobial BCs from patients with bloodstream infection.ResultsThe rapid method herein proposed is based on short-term culture in liquid media allowing selective growth of microorganisms recovered from polymicrobial BCs, followed by rapid identification by MALDI-TOF MS. To evaluate the accuracy of this method, 56 polymicrobial BCs were comparatively analyzed with the rapid and routine methods. The results showed concordant identification for both microbial species in 43/50 (86%) BCs containing two different microorganisms, and for two microbial species in six BCs containing more than two different species. Overall, 102/119 (85.7%) microorganisms were concordantly identified by the rapid and routine methods using a cut-off value of 1.700 for valid identification. The mean time to identification after BC positivity was about 4.2 h for streptococci/enterococci, 8.7 h for staphylococci, 11.1 h for Gram-negative bacteria, and 14.4 h for yeast, allowing a significant time saving compared to the routine method.ConclusionsThe proposed method allowed rapid and reliable microbial identification in polymicrobial BCs, and could provide clinicians with timely, useful information to streamline empirical antimicrobial therapy in critically ill patients.

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Section: Discussionmentioning

confidence: 99%

A new culture-based method for rapid identification of microorganisms in polymicrobial blood cultures by MALDI-TOF MS

et al. 2019

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“…365 monomicrobial growth were correctly identified to the species (72.1%) or genus (89.6%) level. The remaining 40 polymicrobial growth showed 82.5% (33/40) of the isolates were correctly identified to the species level and 92.5% (37/40) to the genus level [ 36 ]. Pretreatments to remove non-bacterial proteins are a prerequisite for the successful application of MALDI-TOF MS in patient-derived BCs.…”

Section: Classification and Identification Of Microorganismsmentioning

confidence: 99%

Current status of MALDI-TOF mass spectrometry in clinical microbiology

Hou

Chiang-Ni

Teng³

2019

Journal of Food and Drug Analysis

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215

137

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Mass spectrometry (MS) is a type of analysis used to determine what molecules make up a sample, based on the mass spectrum that are created by the ions. Mass spectrometers are able to perform traditional target analyte identification and quantitation; however, they may also be used within a clinical setting for the rapid identification of bacteria. The causative agent in sepsis is changed over time, and clinical decisions affecting the management of infections are often based on the outcomes of bacterial identification. Therefore, it is essential that such identifications are performed quickly and interpreted correctly. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometer is one of the most popular MS instruments used in biology, due to its rapid and precise identification of genus and species of an extensive range of Gram-negative and-positive bacteria. Microorganism identification by Mass spectrometry is based on identifying a characteristic spectrum of each species and then matched with a large database within the instrument. The present review gives a contemporary perspective on the challenges and opportunities for bacterial identification as well as a written report of how technological innovation has advanced MS. Future clinical applications will also be addressed, particularly the use of MALDI-TOF MS in the field of microbiology for the identification and the analysis of antibiotic resistance.

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“…Дальнейшие исследования, направленные на сокра щение времени идентификации грибов, стали возмож ными после разработки методов по предварительной обработке положительной гемокультуры с помощью MALDITOF массспектрометрии, минуя этап культивиро вания и ПЦР. Для реализации такого подхода к иден тификации доступны как коммерческие тестсистемы, так и внутренние (in house) алгоритмы, основанные либо на дифференциальном центрифугировании, либо на ис пользовании реагентов для лизиса клеточной стенки грибов (Таблица 4) [16,[19][20][21][22][23].…”

Section: Discussionunclassified

In house method for rapid identification of fungi from fungus-positive bottles by MALDI-TOF mass spectrometry in patients with bloodstream infection

Мальчикова

Klyasova

2022

CMAC

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Objective. To present the results of using in-house method for rapid identification of fungi from funguspositive bottles with routine conventional culture-based identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) in patients with bloodstream infection. Materials and Methods. Prospective study was performed from 2016 to 2019 at the National Research Center for Hematology, Moscow. During the study period, all blood cultures (BC) bottles obtained from hematological patients were incubated in the BACTEC FX system (Becton Dickinson, USA). Positive BC bottles were examined by Gram stain. In house method was used after Gram stain was positive for yeast cells or hyphae. For that, BC media was transfer from fungus-positive bottles into tube. In house method included series section steps consisted from centrifugation and extraction of fungal proteins by adding of sodium dodecyl sulfate. Routine conventional culture-based identification on Sabouraud with chloramphenicol agar (bioMerieux, France) for yeasts and on Sabouraud dextrose agar (Oxoid, UK) for molds was used simultaneously with the in house method. Results. During the study period, 16 fungus-positive bottles were obtained from which were isolated in monoculture 14 (87.5%) Candida spp.: C. parapsilosis (n = 5), C. tropicalis (n = 4), C. albicans (n = 3), C. krusei (n = 1), C. guilliermondii (n = 1), one (6.3%) Rhodotorula mucilaginosa and one (6.3%) Fusarium dimerum. The in house method resulted in 75% (12⁄16) and 68.8% (11⁄16) identification rate at the genus and species level of fungi, respectively. The identification of fungi to species level was confirmed by conventional culture-based method in all cases. The median time from the start of vial incubation in BACTEC FX system to identification of fungi by in house method was less than conventional culture-based identification: 36 hrs 20 min vs 55 hrs 31 min (p = 0.028). Conclusions. A high rate of correct direct species identification and significant reduction in time to verification of fungi from fungus-positive bottles by in house method were obtained. The proposed in house method should be recommended for use in real microbiology practice to reduce the time for submitting results of identification to clinical units.

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Applicability of an in-House Saponin-Based Extraction Method in Bruker Biotyper Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry System for Identification of Bacterial and Fungal Species in Positively Flagged Blood Cultures

Cited by 19 publications

References 23 publications

A new culture-based method for rapid identification of microorganisms in polymicrobial blood cultures by MALDI-TOF MS

A new culture-based method for rapid identification of microorganisms in polymicrobial blood cultures by MALDI-TOF MS

Current status of MALDI-TOF mass spectrometry in clinical microbiology

In house method for rapid identification of fungi from fungus-positive bottles by MALDI-TOF mass spectrometry in patients with bloodstream infection

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