Biotyper System Research Articles

Multicenter Study Demonstrates Standardization Requirements for Mold Identification by MALDI-TOF MS.

ObjectivesRapid and accurate mold identification is critical for guiding therapy for mold infections. MALDI-TOF MS has been widely adopted for bacterial and yeast identification; however, few clinical laboratories have applied this technology for routine mold identification due to limited database availability and lack of standardized processes. Here, we evaluated the versatility of the NIH Mold Database in a multicenter evaluation.MethodsThe NIH Mold Database was evaluated by eight US academic centers using a solid media extraction method and a challenge set of 80 clinical mold isolates. Multiple instrument parameters important for spectra optimization were evaluated, leading to the development of two specialized acquisition programs (NIH method and the Alternate-B method).ResultsA wide range in performance (33–77%) was initially observed across the eight centers when routine spectral acquisition parameters were applied. Use of the NIH or the Alternate-B specialized acquisition programs, which are different than those used routinely for bacterial and yeast spectral acquisition (MBT_AutoX), in combination with optimized instrument maintenance, improved performance, illustrating that acquisition parameters may be one of the key limiting variable in achieving successful performance.ConclusionSuccessful mold identification using the NIH Database for MALDI-TOF MS on Biotyper systems was demonstrated across multiple institutions for the first time following identification of critical program parameters combined with instrument optimization. This significantly advances our potential to implement MALDI-TOF MS for mold identification across many institutions. Because instrument variability is inevitable, development of an instrument performance standard specific for mold spectral acquisition is suggested to improve reproducibility across instruments.

Pseudomonas grimontii, causal agent of turnip bacterial rot disease in Japan

From April to May 2012, a rot disease on turnips grown in greenhouses in southern Hokkaido, Japan caused severe leaf blight, brown discoloration of crown surface, and browning and rot of the interior of main roots. From diseased tissues, bacteria that formed pale beige- to cream-colored colonies were isolated and demonstrated to be pathogenic on turnip after inoculation and reisolation of the isolates. They were gram-negative, aerobic rods with one to three polar flagella, and identified as Pseudomonas grimontii based on biochemical and physiological characterizations, multilocus sequence analysis using concatenated sequences of 16S rRNA, rpoD, gyrB, and rpoB genes, and MALDI-TOF MS analysis using the Bruker MALDI Biotyper system. To the best of our knowledge, P. grimontii has not been isolated previously in Japan, so this study might be the first report of the species in Japan. Although the significance of P. grimontii as a plant pathogenic bacterium has not yet been fully evaluated, we proved in this study that the bacterium is the causal agent of the disease and that its virulence is strong. These findings indicate that it is necessary to record P. grimontii as a novel pathogen posing a threat to plants and requiring vigilance.

MALDI-TOF bacterial subtyping to detect antibiotic resistance

The spread of bacterial resistance has been continuously increasing in the recent decade. Multi-drug resistant (MDR) bacteria now represent one of the most worrisome public health issues, as they seriously complicate the treatment of infections, often leaving few therapeutic options.Enterobacteria and Staphylococcus aureus are among the most common bacterial pathogens, while Bacteroides fragilis is the most frequent anaerobic pathogen. All of these species can cause severe and life-threatening infections, and represent the most frequent causes of antibiotic-resistant healthcare-associated infections worldwide, as they frequently exhibit resistance to various classes of antibiotics. Resistance to carbapenems, the last resort beta-lactam agent, is a particularly threatening problem. Achieved by different mechanisms, leads to total inefficacy of any beta-lactam agent.During the recent years, MALDI-TOF mass spectrometry has become established as the reference method for bacterial identification in routine practice. It has proven to be a reliable and robust method to detect specific peaks in bacterial mass spectra, corresponding to specific resistance markers, enabling the instant detection of resistant isolates in real time during the standard routine identification process. Here, we investigated the performance of the subtyping module of the MALDI Biotyper system (Bruker Daltonik, GmbH) for the instant identification of KPC-producing Klebsiella pneumoniae, methicillin-resistant Staphylococcus aureus, and carbapenemase-producing Bacteroides fragilis during the identification workflow. We evaluated accuracy and potential impact on turnaround time. Furthermore, we investigated the possibility to extend the subtyping for detection of the KPC-specific marker to bacterial species other than K. pneumoniae.

Probiotic Potential and Safety Evaluation of Enterococcus faecalis OB14 and OB15, Isolated From Traditional Tunisian Testouri Cheese and Rigouta, Using Physiological and Genomic Analysis.

Lactic acid bacteria (LAB) strains OB14 and OB15 were isolated from traditional Tunisian fermented dairy products, Testouri cheese and Rigouta, respectively. They were identified as Enterococcus faecalis by the MALDI TOF-MS (matrix assisted laser desorption-ionization time of flight mass spectrometry) biotyper system and molecular assays (species-specific PCR). These new isolates were evaluated for probiotic properties, compared to E. faecalis Symbioflor 1 clone DSM 16431, as reference. The bacteria were found to be tolerant to the harsh conditions of the gastrointestinal tract (acidity and bile salt). They were low to moderate biofilm producers, can adhere to Caco-2/TC7 intestinal cells and strengthen the intestinal barrier through the increase of the transepithelial electrical resistance (TER). Susceptibility to ampicillin, vancomycin, gentamicin and erythromycin has been tested using the broth microdilutions method. The results demonstrated that E. faecalis OB14 and OB15 were sensitive to the clinically important ampicillin (MIC = 1 μg/mL) and vancomycin (MIC = 2 μg/mL) antibiotics. However, Whole Genome Sequencing (WGS) showed the presence of tetracycline resistance and cytolysin genes in E. faecalis OB14, and this led to high mortality of Galleria Mellonella larvae in the virulence test. Hierarchical cluster analysis by MALDI TOF-MS biotyper showed that E. faecalis OB15 was closely related to the E. faecalis Symbioflor 1 probiotic strain than to OB14, and this has been confirmed by WGS using the average nucleotide identity (ANI) and Genome-to-Genome Hybridization similarity methods. According to these results, E. faecalis OB15 seems to be reliable for future development as probiotic, in food or feed industry.

A study of bacterial distribution and drug resistance in skin and soft tissue infection

Objective: To investigate pathogenic bacteria and drug resistance in the patients with skin and soft tissue infection in order to provide the scientific evidences for clinical reasonable use of antibiotics. Methods: A retrospective analysis was performed on patients with skin and soft tissue infections in Department of Dermatology, Peking University Third Hospital from January 2012 to December 2017. Pus, secretions, skin lesions, urine, throat swabs, and alveolar lavage fluid were collected for bacterial culture, bacterial species were identified by VITEK2 Compact system and BD-Bruker MALDI Biotyper system. Drug resistance was detected by K-B agar diffusion method recommended by CLSI. Results: A total of 392 strains of bacteria were isolated from 327 patients distributed in 21 genus and 56 species, of which 225 were gram-positive cocci (57.40%), 114 were gram-negative rods (29.08%), 46 were gram-positive rods (11.73%), and 7 were gram-negative cocci (1.79%). The top 3 bacteria were Staphylococcus aureus 91(23.21%), Staphylococcus epidermidis 42 (10.71%), and Pseudomonas aeruginosa 24 (6.12%). Staphylococcus had a high rate of resistance to penicillin and erythromycin (>50%). Gram-negative rods were resistant to ampicillin (86.1%), and also had certain resistance to most second-generation and some third-generation cephalosporin (about 50%). There was no significant change in the drug resistance rate of MRSA compared to MSSA. Only the resistance rate to tetracycline was statistically different (P<0.05). Conclusion: The emergence of drug-resistant strains is an important factor leading to refractory infections. There are a wide range of pathogenic bacteria species among the skin and soft tissue infection patients, and antimicrobial drugs should be chosen wisely according to drug sensitivity.

Performance of matrix assisted laser desorption ionization time of flight biotyper system in clinical bacteria identification

Objective: To evaluate the performance of MALDI Biotyper system in identification of clinically isolated pathogens so as to provide a new rapid identification method. Methods: Total 21 270 pathogens strains, isolated from the First Affiliated Hospital of Fujian Medical Universityduring Nov. 2015 to Dec. 2016, were identified by VITEK-Ⅱ, API and MALDI Biotyper system, respectively.The isolated strains were confirmed by DNA sequencing. Results: The identification of common bacteria with MALDI Biotyper and phenotypic system is highly consistent (>95% and >90%). Among 43 strains of anaerobic bacteria, MALDI Biotyper could identify 90.7% bacteria to species level and 97.7% bacteria to genus level with the statistical significance(χ(2)=6.76, P<0.01), while phenotypic system only identified 65.1% bacteria to species and 69.8% bacteria to genus. Also, no statistical significance was shown for Trichosporon and Candida(P>0.05). MALDI Biotyper could identify 76% filamentous fungi and all of Actinomycetes, Nocardia, Mycobacterium and Legionella to genus level. Conclusions: MALDI Biotyper is an easy-performed, sensitive method for the identification of clinically isolated pathogens. Additionally, the pretreatment and reference database has the effect on identification.

A Full MALDI-Based Approach to Detect Plasmid-Encoded KPC-Producing Klebsiella pneumoniae.

KPC-producing Klebsiella pneumoniae represents a severe public health concern worldwide. The rapid detection of these isolates is of fundamental importance for the adoption of proper antibiotic treatment and infection control measures, and new applications of MALDI-TOF MS technology fit this purpose. In this study, we present a full MALDI-based approach to detect plasmid-encoded KPC-producing strains, accomplished by the automated detection of a KPC-specific peak (at 11,109 m/z) by a specific algorithm integrated into the MALDI Biotyper system (Bruker Daltonik), and the confirmation of carbapenemase activity by STAR-Carba imipenem hydrolysis assay. A total of 6209 K. pneumoniae isolates from Italy and Germany were investigated for the presence of the KPC-related peak, and a subset of them (n = 243) underwent confirmation of carbapenemase activity by STAR-Carba assay. The novel approach was further applied directly to positive blood culture bottles (n = 204), using the bacterial pellet obtained with Sepsityper kit (Bruker Daltonik). The novel approach enabled a reliable and very fast detection of KPC-producing K. pneumoniae strains, from colonies as well as directly from positive blood cultures. The automated peak detection enabled the instant detection of KPC-producing K. pneumoniae during the routine identification process, with excellent specificity (100%) and a good sensitivity (85.1%). The sensitivity is likely mainly related to the prevalence of the specific plasmid harboring clones among all the KPC-producing circulating strains. STAR-Carba carbapenemase confirmation showed 100% sensitivity and specificity, both from colonies and from positive blood cultures.

Improvement of a rapid direct blood culture microbial identification protocol using MALDI-TOF MS and performance comparison with SepsiTyper kit

Fast diagnosis of pathogens is critical to guarantee the most adequate therapy for infections; bacterial culture methods, which constitute the actual gold standard, are precise and sensitive but rather slow. Today, new methods have been made available to enable faster diagnosis, with the Matrix-Assisted Laser Desorption Ionization–Time Of Flight Mass Spectrometry (MALDI-TOF MS) technique being the most promising. Even if simpler and faster than traditional bacterial culture methods, analysis of positive blood cultures via MALDI-TOF MS requires a preliminary extraction process of samples. In this study, we compared two extraction protocols for bacterial identification directly from positive blood cultures using the Bruker MALDI Biotyper system (Bruker Daltonics, Billerica, MA). In particular, we evaluated the time employed and the overall performance for their accurate identification. In this work, the performances of a commercial extraction kit, named SepsiTyper™ Kit, and those of the protocol developed by Treibmann et al. were evaluated and proven to be similar. However, the SELTERS method represents the best compromise price/performance. Lastly, an in-house developed analysis protocol has been tested, and the introduced optimizations granted a performance level equal if not better than the SepsiTyper kit, a reduced processing time and reduced costs.

Analysis of adult damselfly fecal material aids in the estimation of antibiotic-resistant Enterobacterales contamination of the local environment.

Because damselflies are ubiquitously but focally present in natural environments and play a critical role as predators of other insect species, the fecal matter of damselflies may be useful for investigating antibiotic-resistant bacterial populations, including human pathogens, in local environments. We therefore examined the prevalence of antibiotic-resistant bacteria, including Enterobacterales, in fecal material from 383 damselflies (adults and larvae) collected from seven locations around Sapporo City, Japan, in 2016 and 2017. Fecal samples were plated on soybean casein digest (SCD) agar plates with and without antibiotics (SCD-A and SCD-w/o, respectively) to identify environmental bacteria and gut bacteria, respectively, and on MacConkey agar plates with antibiotics (MacConkey-A) to select for Gram-negative bacteria, including human pathogenic Enterobacterales species. The prevalence of colonies on each of the plates was compared, and representative colonies on MacConkey-A plates were identified to the species level using an API 20E kit and the MALDI Biotyper system. Overall, SCD-w/o plates showed a gut bacterial load of approximately 108 colony-forming units per adult damselfly or larva. There was a significant difference between the prevalence of colonies on the SCD-A and MacConkey-A plates, and a significantly increased prevalence of antibiotic-resistant bacteria on MacConkey-A plates was observed in samples collected from Shinoroshinkawa. Cluster analysis based on minimum inhibitory concentration values of 59 representative isolates from MacConkey-A agar plates revealed that samples from Shinoroshinkawa contained a higher prevalence of Enterobacterales than those from other sampling locations. Thus, fecal materials discharged by adult damselflies could be used in future studies as a simple tool for estimating antibiotic-resistant bacteria, including Enterobacterales species, in the local environment.

Identification of the Slovak traditional cheese “Parenica” microflora

Numerous studies have demonstrated the higher accuracy, faster time-to-results and lower costs provided by MALDI Biotyper systems compared to classical methods. In this study, the culturable population of total count of bacteria, enterococci, coliforms bacteria, lactic acid bacteria (LAB) and microscopic fungi and yeasts from cow’s dairy products was identified using the MALDI-TOF MS Biotyper. Altogether, 50 samples of the Slovak cheese “Parenica” were examined. Total numbers of bacteria were cultured on Plate count agar at 37 °C for 24–48 h, aerobically; enterococci were cultured on Enterococcus selective agar at 37 °C for 24–48 h, aerobically; coliforms bacteria were cultured on Violet Red Bile lactose agar at 37 °C for 24–48 h, aerobically. The LAB were cultured on MRS (Main Rogosa agar), MSE and APT agar at 30 °C in microaerophilic conditions. The microscopic fungi and yeasts were cultured on Malt extract agar at 25 °C for 5 days, aerobically. Isolated strains (total 669) were subjected to identification by the MALDI-TOF MS. Among total count the identified bacteria mostly were Acinetobacter baumannii, Bacillus cereus, Micrococcus luteus and Staphylococcus warneri. Escherichia coli and Enterobacter cloacae were the most abundant coliform bacteria representatives identified. Coliform bacteria included Citrobacter, Hafnia and Klebsiella. Altogether three genera belonged to the LAB – Lactobacillus, Lactococcus and Leuconostoc were identified with Lactococcus lactis, Lactobacillus plantarum, Lactobacillus coryniformis, L. fructivorans and Leuconostoc mesenteroides were considered as the dominated LAB species in dairy products. Among yeasts, Kluyveromyces lactis, Candida zeylanoides and Yarrowia lipolytica were among the most isolated.

Application of the biocopolymer preparation system, rapid BACpro® II kit, for mass-spectrometry-based bacterial identification from positive blood culture bottles by the MALDI Biotyper system

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used for identification of microorganisms from positive blood cultures. Pretreatments to effectively remove non-bacterial components and selectively collect microorganisms are a prerequisite for successful identification, and a variety of home-brew and commercial protocols have been reported. Although commercially available kits, mainly the Sepsityper Kit, are increasingly used, the identification rates reported often are not satisfactory, particularly for Gram-positive isolates. We recently developed a method to collect bacteria from positive blood culture bottles using a polyallylamine-polystyrene copolymer that has been used in wastewater processing. This pretreatment protocol is now commercially available as the rapid BACpro® II kit (Nittobo Medical Co., Tokyo, Japan). The operation time required for processing using this novel kit is approximately 10 min, and the entire procedure can be completed within a biosafety cabinet.Since the performance of the rapid BACpro® II kit has not been tested using the MALDI Biotyper system, we prospectively evaluated the performance of the rapid BACpro® II kit as compared with the Sepsityper® kit.Performance of the rapid BACpro® II kit was evaluated using a total of 193 monomicrobial cases of positive blood culture. Medium from blood culture bottles was pretreated by the rapid BACpro® II kit or the Sepsityper® Kit, and isolated cells were subjected to direct identification by MS fingerprinting in parallel with conventional subculturing for reference identification. The overall MALDI-TOF MS-based identification rates with >1.7 score and >2.0 score obtained using the rapid BACpro® II kit were 99.5% and 80.8%, respectively, whereas those obtained using the Sepsityper® Kit were 89.1% and 68.4%, respectively (P < 0.05 for >1.7 and P < 0.05 for >2.0 by Pearsons's chi-square). In Gram-positive cases, the rapid BACpro® II kit gave identification rate of 100% with >1.7 score and 69.4% with >2.0 score, whereas there were 84.7% and 56.8%, respectively by the Sepsityper® Kit (P < 0.05 for >1.7).These results are preliminary, but considering that this new kit is easy to perform and the identification rates are promising, the rapid BACpro® II kit deserves assessment in a larger-scale study with a variety of platforms for MS-based bacterial identification.

Routine use of MALDI-TOF MS for anaerobic bacterial identification in clinical microbiology

In 2013, we adopted MALDI-TOF MS using the Bruker Biotyper system for identification of anaerobic bacteria into our routine clinical practice. Here, we describe our experience with the use of MALDI-TOF MS for anaerobic bacterial identification, highlighting its value in replacing the more costly and time-consuming 16S ribosomal RNA gene PCR plus sequencing-based approach as the primary method of anaerobic bacterial identification. We also describe our more recent experience with the use of early/rapid MALDI-TOF MS for identification of anaerobic bacteria performed on short incubation (4–6 h) plated aerobic media from anaerobic blood culture bottles positive for Gram-negative bacilli.

Evaluation of the Vitek MS and the MALDI Biotyper systems for the identification of less commonly isolated but clinically relevant anaerobes and facultative anaerobes

The Vitek MS and MALDI Biotyper systems were evaluated for the identification of 221 strains belonging to less commonly isolated anaerobes and facultative anaerobes. Identifications were performed using direct deposit (DD), on-target extraction (FA) and full extraction (EXT). After DD, 29.9% and 34.3% of Gram-positive isolates (n = 137) as well as 36.9% and 58.3% of Gram-negative isolates (n = 84) were identified at the species-level with the Vitek-MS and the Biotyper system, respectively. The rates of correct species identification with the Biotyper system were significantly increased after extraction for Gram-positive isolates (FA: 75.2%, EXT: 78.1%) and Gram-negative isolates (FA: 72.6%, EXT: 78.6%). Extraction permitted to achieve higher correct species identification rates only for Gram-positive isolates (FA: 35.8%, EXT: 36.5%) with the Vitek MS. Thus, the accuracy of both systems needs to be further increased by expanding the databases. In the meantime, we recommend using a preliminary extraction step to obtain reliable results at least for the identification of Gram positive anaerobic bacteria with both systems.

Performance of mass spectrometric identification of clinical Prevotella species using the VITEK MS system: A prospective multi-center study

Prevotella species, members of the human microbiota, can cause opportunistic infections. Rapid and accurate identification of Prevotella isolates plays a critical role in successful treatment, especially since the antibiotic susceptibility profile differs between species. Studies, mostly carried out using the Matrix Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) Biotyper system, showed that MALDI-TOF MS is an accurate, rapid and satisfactory method for the identification of clinically important anaerobes. In this multi-center study, we assessed the performance of the MALDI-TOF MS VITEK MS system for the identification of clinical Prevotella isolates. A total of 508 Prevotella isolates, representing 19 different species, collected from 11 European countries, Kuwait and Turkey between January 2014 and April 2016, were identified using VITEK MS (v3.0). The reliability of the identification was assessed by 16S rRNA gene sequencing. Using VITEK MS, 422 (83.1%) of the 508 isolates were identified on the species level, 459 (90.4%) on the genus level. A total of 49 (9.6%) isolates were not identified correctly. 16S rRNA gene sequencing results showed that this was partly due to the fact that several species were not represented in the database. However, some species that were represented in the database were also not identified. Five Prevotella strains were misidentified at the genus level, 2 of these strains belonged to a species not represented in the database. In general, the VITEK MS offers a reliable and rapid identification of Prevotella species, however the databases needs to be expanded.

ネコ尿検体に対してMALDI Bio Typer Systemを応用した直接菌種同定の検討

ネコ尿検体に対してMALDI Bio Typer Systemを応用した直接菌種同定の検討

Application of MALDI-TOF MS Systems in the Rapid Identification of Campylobacter spp. of Public Health Importance

Campylobacteriosis is an infectious gastrointestinal disease caused by Campylobacter spp. In most cases, it is either underdiagnosed or underreported due to poor diagnostics and limited databases. Several DNA-based molecular diagnostic techniques, including 16S ribosomal RNA (rRNA) sequence typing, have been widely used in the species identification of Campylobacter. Nevertheless, these assays are time-consuming and require a high quality of bacterial DNA. Matrix-assisted laser desorption ionization (MALDI) time-of-flight (TOF) MS is an emerging diagnostic technology that can provide the rapid identification of microorganisms by using their intact cells without extraction or purification. In this study, we analyzed 24 American Type Culture Collection reference isolates of 16 Campylobacter spp. and five unknown clinical bacterial isolates for rapid identification utilizing two commercially available MADI-TOF MS platforms, namely the bioMérieux VITEK® MS and Bruker Biotyper systems. In addition, 16S rRNA sequencing was performed to confirm the species-level identification of the unknown clinical isolates. Both MALDI-TOF MS systems identified the isolates of C. jejuni, C. coli, C. lari, and C. fetus. The results of this study suggest that the MALDI-TOF MS technique can be used in the identification of Campylobacter spp. of public health importance.

Comprehensive Evaluation of the MBT STAR-BL Module for Simultaneous Bacterial Identification and β-Lactamase-Mediated Resistance Detection in Gram-Negative Rods from Cultured Isolates and Positive Blood Cultures.

Objective: This study evaluated the capability of a MALDI Biotyper system equipped with the newly introduced MBT STAR-BL module to simultaneously perform species identification and β-lactamase-mediated resistance detection in bacteremia -causing bacteria isolated from cultured isolates and patient-derived blood cultures (BCs).Methods: Two hundred retrospective cultured isolates and 153 prospective BCs containing Gram-negative rods (GNR) were collected and subjected to direct bacterial identification, followed by the measurement of β-lactamase activities against ampicillin, piperacillin, cefotaxime, ceftazidime, and meropenem using the MBT STAR-BL module. The results and turnaround times were compared with those of routine microbiological processing. All strains were also characterized by beta-lactamase PCR and sequencing.Results: Using the saponin-based extraction method, MALDI-TOF MS correctly identified bacteria in 116/134 (86.6%) monomicrobial BCs. The detection sensitivities for β-lactamase activities against ampicillin, piperacillin, third-generation cephalosporin and meropenem were 91.3, 100, 97.9, and 100% for cultured isolates, and 80.4, 100, 68.8, and 40% for monomicrobial BCs (n = 134) respectively. The overall specificities ranged from 91.5 to 100%. Furthermore, the MBT STAR-BL and conventional drug susceptibility test results were concordant in 14/19 (73.7%) polymicrobial cultures. Reducing the logRQ cut-off value from 0.4 to 0.2 increased the direct detection sensitivities for β-lactamase activities against ampicillin, cefotaxime and meropenem in BCs to 85.7, 87.5, and 100% respectively. The MBT STAR-BL test enabled the reporting of β-lactamase-producing GNR at 14.16 and 47.64 h before the interim and final reports of routine BCs processing, respectively, were available.Conclusion: The MALDI Biotyper system equipped with the MBT STAR-BL module enables the simultaneous rapid identification of bacterial species and β-lactamase-mediated resistance from BCs and cultured isolates. Adjustment of the logRQ cut-off value to 0.2 significantly increased the detection sensitivities for clinically important drug-resistant pathogens.

Application of the MALDI Biotyper to clinical microbiology: progress and potential

ABSTRACTIntroduction: The introduction of the MALDI Biotyper in laboratories substantially changed microbiology practice, this has been called a revolution. The system accelerated diagnostic while costs were reduced and accuracy was increased. In just a few years MALDI-TOF MS became the first-line identification tool for microorganisms. Ten years after its introduction, more than 2000 MALDI Biotyper systems are installed in laboratories which are performing routine diagnostic, and the number is still increasing.Areas covered: This article summarises changes in clinical microbiology introduced by the MALDI Biotyper and its effects, as it has been published in peer reviewed articles found in PubMed. Further, the potential of novel developments to increase the value of the system is described.Expert commentary: The MALDI Biotyper has significantly improved clinical microbiology in the area of microorganism identification. Now new developments and applications, e.g. for typing and resistance testing, might further increase its value in clinical microbiology. The systems might get the central diagnostic analyser which is getting integrated into the widely automated microbiology laboratories of the future.

Antimicrobial Resistant Enterococcus faecium, Enterococcus faecalis, and Other Enterococcus Species Isolated From Foal Feces in Japan

Antimicrobial resistant (AMR) Enterococci are an emerging problem for human and animal health. Few studies have focused on the detection of AMR in foals; therefore, we conducted this study to investigate the occurrence of AMR Enterococci in foal feces in Japan. A total of 72 fecal samples from healthy foals were collected from seven horse stables, in Hokkaido Prefecture, from June 2015 to January 2016. Enterococci were isolated and identified using EF Agar Base selective media, and the species confirmed using MALDI Biotyper systems. The minimum inhibitory concentration of nine antimicrobial agents for the isolates was determined based on the Clinical Laboratory Standard Institute guidelines. A total of 183 isolates of Enterococci were identified as Enterococcus faecium 54.1% (99/183), Enterococcus faecalis 16.4% (30/183), and other species 29.5% (54/183). The highest occurrence of E. faecium antibiotic resistant was to erythromycin at 55.6% (55/99), followed by enrofloxacin, kanamycin, and oxytetracycline, at 30.3%, 7.1%, and 4.0%, respectively. In contrast, E. faecalis isolates showed higher resistant to oxytetracycline 76.7%, kanamycin 46.7%, gentamycin 30.0%, chloramphenicol 16.7%, lincomycin 30.0%, and tylosin 30.0%. E. faecium highly resistant to erythromycin and enrofloxacin but lowly resistant to gentamycin and tylosin, and E. faecalis was highly resistant to kanamycin, oxytetracycline, and lincomycin. Antimicrobial resistant E. faecalis (30.0%) and E. faecium (4.0%) isolates show multidrug resistant to at least three different classes of antibiotics. Our study indicated that foal feces may act as a source of AMR genes for Enterococci that may be transmitted to other animals, humans, and the environment.

Use of the MALDI BioTyper system and rapid BACpro with MALDI-TOF MS for rapid identification of microorganisms causing bacterial urinary tract infection in feline urine samples.

With the increasing number of cats kept as pets, opportunities to treat cats with lower urinary tract disease (LUTD) have recently increased in the clinical veterinary field. Urine samples collected from 50 cats with bacterial cystitis brought to Maeda Veterinary Hospital between August 10, 2015 and March 31, 2017 were used in the study. Sample preparation of the urine was performed using a MALDI Sepsityper kit and rapid BACpro. To identify the isolates, MALDI-TOF MS was performed on an AutoFlex TOF/TOF mass spectrometer. MALDI-TOF MS using rapid BACpro for pretreatment was found to be a quick and reliable method for identification of bacteria from infected urine, with a shortened analysis time enabling earlier and more accurate selection of antibiotics for treatment of feline LUTD.