2.16-2.17收获：VITEK MS 全自动快速微生物质谱检测系统（MALDI-TOF）

细菌耐药

近两日在UCLA学习中发现一种可用于临床细菌鉴定的快速检测技术，为VITEK® MS 全自动快速微生物质谱检测系统。该技术可在半小时内完成近百份标本的检测。该技术已在欧洲和日本用于临床细菌鉴定四年多，取得了很好的临床应用评价。美国由于FDA的管理比较严格，直到现在还没有用于临床。我所在学习的UCLA临床微生物学实验室恰好正在开展该技术方法的临床前验证，这边的director也很认可该技术。今天早上我特地一大早起来，看了一下他们使用该技术进行细菌鉴定的全过程，确实很方便。

我权衡了一下，该技术用于临床细菌鉴定可以在1天内拿到结果，优点是快速和芯片成本低（具体还不清楚，这边的director介绍时说每株细菌的成本为pennies），缺点是仪器设备贵，约25万美元左右。不过从长远来说，还是节约成本的，因为耗材很便宜。此外，该技术也可以直接对临床标本进行鉴定，以及药敏，不过还处于科研阶段，我已检索到相关的文献，很多。相信，该技术将会是临床微生物学领域内很有应用前景的一项技术。

细菌耐药

VITEK® MS 全自动快速微生物质谱检测系统利用创新的质谱技术MALDI-TOF（基质辅助激光解吸电离飞行时间）数分钟内提供鉴定结果。

通过VITEK® MS Prep Station软件平台，多位检验技师可以在集中或者分散的工作平台同步处理标本板，VITEK MS可以同机检测4个标本板，一个标本板含有48个孔位，故每次检测最大通量为192个样本。

含有条形码的标本板灵活易用，可直接扫描输入数据，实现检测过程中数据追踪。可直接在标本上处理样品。


样品处理只需两步：

1 - 直接将细菌涂布在标本板上，然后加上基质；

2 - 将标本板放入VITEK MS仪器，几分钟内显示鉴定结果。





安全性及溯源性
利用VITEK® MS准备工作站，每个样品的信息都能和标本板上的孔位以及VITEK 2卡片一一对应，并实现一站式鉴定及药敏检测。平板培养基、基质和标本板的条码唯一性保证了病人检测信息的可追溯性。


准确鉴定结果
数据库包含大量临床相关细菌菌种。运用专用计算方法增加细菌鉴定的准确率。

细菌鉴定的过程是先通过MALDI-TOF基质辅助激光解吸电离飞行时间获得图谱，然后与VITEK MS数据库中不同微生物家族的种/属特定图谱相比对，从而得出鉴定结果。

该系统还包含了宽泛灵敏的质量检测范围（大于1万道尔顿），拥有高分辨率的质谱信号，从而进一步提高检测能力。



VITEK MS 获得欧洲体外认可用于临床常规检测。(CE marked)

VITEK MS & VITEK 2 & Myla™ – 一站式鉴定及药敏解决方案
VITEK MS 连接市场上最可靠VITEK 2药敏系统，通过创新中端软件 (Myla™)，提供一站式鉴定及药敏解决方案。



灵活高效的Myla™：


协助完成鉴定、药敏检测  
提供远程访问和维护
确保一站式报告结果

您可以在户外或者出差时，通过智能电话等设备，在Myla平台上远程验证结果。所有结果把握在你的指尖中. (Click to enlarge)



源于微生物学家，服务于微生物学家
生物梅里埃是唯一一家能够提供微生物鉴定和药敏一站式解决方案的公司，且开发了全新中端软件 (Myla™) 。这一设计理念源于微生物学家，且忠实服务于微生物学家。作为微生物领域的引领者，我们深刻理解您的需求，我们将致力于提高您的实验室生产力和病人信息管理能力。

细菌耐药

在网上查了一下，国内已经有不少医院在招标采购该设备了。临床微生物领域即将迎来一场空前的技术革命，临床微生物学的快速诊断时代即将到来，让我们拭目以待吧！

细菌耐药

J Clin Microbiol. 2012 Feb 8. [Epub ahead of print]
Comparison of the Microflex LT and Vitek(R) MS systems for the routine identification of bacteria by Matrix-Assisted Laser Desorption-Ionization Time-Of-Flight Mass Spectrometry.
Martiny D, Busson L, Wybo I, Ait El Haj R, Dediste A, Vandenberg O.
SourceDepartment of Microbiology, Saint-Pierre University Hospital & Jules Bordet Institute, Brussels, Belgium.

Abstract
This study compares the performance of three Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) systems: Microflex LT (Bruker Daltonics, Bremen, Germany), Vitek MS RUO (Axima Assurance - Saramis database; bioMérieux, Marcy l'Etoile, France), and Vitek MS IVD (bioMérieux).A total of 1,129 isolates including 1,003 routine isolates, 73 anaerobes, and 53 bacterial enteropathogens were tested on the Microflex LT and Axima Assurance devices. The spectra were analyzed using three databases: Biotyper (Bruker Daltonics), Saramis, and Vitek MS (bioMérieux).Among the routine isolates requiring identification to the species level (n=986), 92.7% and 93.2% were correctly identified by the Biotyper and Vitek MS databases, respectively. The Vitek MS database is more specific for the identification of S. viridans. For the anaerobes, the Biotyper database often only identified Fusobacterium isolates to the genus level, which is of low clinical significance, whereas 20% of the Bacteroides species were not identified or were misidentified by the Vitek MS database. For the enteropathogens, the poor discrimination between E. coli and Shigella explains the high proportion of unidentified organisms. In contrast to the Biotyper database, the Vitek MS database properly discriminated all of the S. typhi isolates (n=5). The performance of the Saramis database was globally poorer.In conclusion, for routine procedures, the Microflex LT and Vitek-MS systems are equally good choices in terms of their analytical efficiency. Other factors, including price, workflow and lab activity, will affect the choice of a system.

细菌耐药

J Med Microbiol. 2012 Mar;61(Pt 3):339-44. Epub 2012 Jan 23.
Direct identification of bacteria in urine samples by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and relevance of defensins as interfering factors.
Köhling HL, Bittner A, Müller KD, Buer J, Becker M, Rübben H, Rettenmeier AW, Mosel F.
Source1Institute of Hygiene and Occupational Medicine, University Hospital Essen, University of Duisburg-Essen (Germany), Virchowstraße 171, D-45147 Essen, Germany.

Abstract
Standard methods for the identification of uropathogens that are based on the determination of metabolic activity require cultivation on agar plates, which often takes more than 1 day. If microbial growth on agar plates is slow, or if metabolic activity is impaired by adverse interactions resulting from the patient's condition or from medical treatment, the application of standard methods may lead to delayed or erroneous identification of bacteria. In recent studies, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven to be able to rapidly identify bacteria obtained from cultures. We tested the applicability of this analytical technique for the rapid identification of bacteria collected directly from urine samples and compared the results with those of conventional identification methods, such as the Vitek system, the MicroScan WalkAway system and the API system, and in some cases with the gas chromatographic determination of the bacterial long-chain fatty acid pattern. We analysed a total of 107 urine samples with bacterial counts ranging from 10(2) to ≥10(5) c.f.u. ml(-1). Mass spectrometric identification of bacteria was accomplished for 62 of these samples. In the mass spectra obtained from 40 of the 45 urine samples for which no identification result was achieved, a triplet of very intense peaks corresponding to the human α-defensins 1, 2 and 3 occurred at m/z values of around 3440 Da. This signal suppressed the intensity of the bacterial protein peaks and thus impaired database matching. Our results show that MALDI-TOF MS allows the reliable direct identification of bacteria in urine samples at concentrations as low as 10(3) c.f.u. ml(-1). In a subset of samples, human defensins may occur and impair the mass spectrometric identification of bacteria.

细菌耐药

J Clin Microbiol. 2012 Feb 8. [Epub ahead of print]

Comparison of the Microflex LT and Vitek(R) MS systems for the routine identification of bacteria by Matrix-Assisted Laser Desorption-Ionization Time-Of-Flight Mass Spectrometry.

Martiny D, Busson L, Wybo I, Ait El Haj R, Dediste A, Vandenberg O.

Source

Department of Microbiology, Saint-Pierre University Hospital & Jules Bordet Institute, Brussels, Belgium.

Abstract

This study compares the performance of three Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) systems: Microflex LT (Bruker Daltonics, Bremen, Germany), Vitek MS RUO (Axima Assurance - Saramis database; bioMérieux, Marcy l'Etoile, France), and Vitek MS IVD (bioMérieux).A total of 1,129 isolates including 1,003 routine isolates, 73 anaerobes, and 53 bacterial enteropathogens were tested on the Microflex LT and Axima Assurance devices. The spectra were analyzed using three databases: Biotyper (Bruker Daltonics), Saramis, and Vitek MS (bioMérieux).Among the routine isolates requiring identification to the species level (n=986), 92.7% and 93.2% were correctly identified by the Biotyper and Vitek MS databases, respectively. The Vitek MS database is more specific for the identification of S. viridans. For the anaerobes, the Biotyper database often only identified Fusobacterium isolates to the genus level, which is of low clinical significance, whereas 20% of the Bacteroides species were not identified or were misidentified by the Vitek MS database. For the enteropathogens, the poor discrimination between E. coli and Shigella explains the high proportion of unidentified organisms. In contrast to the Biotyper database, the Vitek MS database properly discriminated all of the S. typhi isolates (n=5). The performance of the Saramis database was globally poorer.In conclusion, for routine procedures, the Microflex LT and Vitek-MS systems are equally good choices in terms of their analytical efficiency. Other factors, including price, workflow and lab activity, will affect the choice of a system.

细菌耐药

Jpn J Infect Dis. 2011;64(4):327-9.
Rapid identification of Cardiobacterium hominis by MALDI-TOF mass spectrometry during infective endocarditis.
Wallet F, Loïez C, Decoene C, Courcol R.
SourceUniversity of Lille Nord de France, Lille, France. wallet@chru-lille.fr

Abstract
We report a new case of Cardiobacterium hominis endocarditis identified during an acute coronary syndrome. The positivity of the blood cultures was confirmed rapidly (50 h) as a result of improvements to the automated detection system, whereby it is no longer necessary to incubate the vials for long periods of time when Aggregatibacter-Cardiobacterium-Eikenella-Kingella infections is suspected. The phenotype-based VITEK 2 NH identification system is not able to distinguish between the two species of Cardiobacterium, as it does not contain C. valvarum in its library. The method for 16S rRNA gene sequence analysis is able to separate the two species but is not available in all laboratories. We used MALDI-TOF mass spectrometry, as an alternative, to rapidly distinguish between C. hominis and C. valvarum, because both species are contained in the system library.

附上全文：

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细菌耐药

Microb Drug Resist. 2011 Sep;17(3):433-42. Epub 2011 May 13.
Molecular and epidemiological analysis of nosocomial carbapenem-resistant Klebsiella spp. using repetitive extragenic palindromic-polymerase chain reaction and matrix-assisted laser desorption/ionization-time of flight.
Treviño M, Navarro D, Barbeito G, García-Riestra C, Crespo C, Regueiro BJ.
SourceServicio de Microbiología, Complejo Hospitalario Universitario de Santiago de Compostela, Santiago de Compostela, Spain. maria.mercedes.trevino.castellano@sergas.es

Abstract
INTRODUCTION: Infections with carbapenem-resistant enterobacteria are an emerging threat. This study reports the microbiologic, clinical, and epidemiologic features and the therapeutic outcomes of the infections caused by carbapenem- and pandrug-resistant Klebsiella emerged in our hospital. Fingerprinting analyses by automated repetitive extragenic palindromic-polymerase chain reaction (rep-PCR) and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry are also compared.

MATERIALS AND METHODS: Carbapenem-resistant Klebsiella spp. affecting 13 patients were investigated using automated rep-PCR (DiversiLab System) and MALDI-TOF. Species identification was performed by Vitek 2 System and MALDI-TOF. Antimicrobial susceptibility testing was made using Vitek 2 System and Etest. Screening for extended spectrum beta-lactamase (ESBL) and carbapenemase production was made by double disk synergy and Hodge tests, respectively. Synergy studies were performed using Etest. DNA array was used for detection of KPC and ESBLs. bla(VIM-1) gene was amplified by PCR and sequencing. Use of carbapenems in the hospital was studied.

RESULTS: A total of 13 patients were found to be colonized/infected with carbapenem-resistant Klebsiella. All patients were previously submitted to surgery and/or presented with severe underlying disease. After carbapenem-resistant Klebsiella isolation, the majority of the patients were treated with amikacin plus carbapenem, tigecycline, or fosfomycin. All Klebsiella isolates (n = 14), except two, had the bla(VIM-1) gene and all Klebsiella pneumoniae also had bla(SHV) gene associated with ESBL production. DiversiLab system showed higher discriminatory power than MALDI-TOF for strain typing.

CONCLUSIONS: The risk of a rapid dissemination and the persistence of these multidrug-resistant strains through the time determine the need to implement routine procedures for metallo-beta-lactamase detection and measures for prevention of the spread of these microorganisms. The combined use of MALDI-TOF for species identification and DiversiLab System for clonal strain typing may be a useful tool for fast and accurate management of nosocomial outbreaks. The potential clinical utility of fosfomycin in this matter should be considered in future studies.

细菌耐药

Eur J Clin Microbiol Infect Dis. 2011 Dec;30(12):1579-86. Epub 2011 Apr 21.
Revisited distribution of nonfermenting Gram-negative bacilli clinical isolates.
Jacquier H, Carbonnelle E, Corvec S, Illiaquer M, Le Monnier A, Bille E, Zahar JR, Beretti JL, Jauréguy F, Fihman V, Tankovic J, Cattoir V.
SourceService de Bactériologie-Virologie, Groupe Hospitalier Lariboisière-Fernand Widal, Assistance Publique-Hôpitaux de Paris, 2 rue Ambroise Paré, 75475 Paris Cedex 10, France. herve.jacquier@lrb.aphp.fr

Abstract
Nonfermenting Gram-negative bacilli (NF-GNB) are ubiquitous environmental opportunistic bacteria frequently misidentified by conventional phenotypic methods. The aim of this study was to determine the distribution of NF-GNB species by 16 S rRNA gene sequencing (used as reference method) and to compare performances of biochemical tests and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). From nine French hospitals, 188 NF-GNB isolates (except P. aeruginosa and A. baumannii) were prospectively collected from 187 clinical samples between December 2008 and May 2009. By using the genotypic approach, 173 (92%) and 188 (100%) isolates were identified to the species and genus level, respectively. They covered 35 species and 20 genera, with a predominance of Stenotrophomonas maltophilia, Achromobacter xylosoxidans, and Pseudomonas putida group bacteria. Of the 173 species-level identified strains, concordant identification to the species-level was obtained for 75.1%, 83% and 88.9% of isolates with API 20 NE strip, the VITEK-2 (ID-GN card) system and MALDI-TOF-MS, respectively. By excluding S. maltophilia isolates accurately identified by the three methods, genus-level identification was much higher for MALDI-TOF-MS (92.9%), compared with API 20 NE and VITEK-2 (76.2% and 80.8%, respectively). In conclusion, MALDI-TOF-MS represents a rapid, inexpensive, and accurate tool for routine identification of NF-GNB in human clinical samples.

细菌耐药

众所周知，真菌的鉴定在临床上比较困难，Vitek对真菌鉴定也没有什么好办法。下面这篇文章谈到了将该技术用于真菌的检测，效果不错。

Mol Biosyst. 2011 Mar;7(3):620-9. Epub 2010 Oct 21.
MALDI-TOF mass spectrometry proteomic phenotyping of clinically relevant fungi.
Putignani L, Del Chierico F, Onori M, Mancinelli L, Argentieri M, Bernaschi P, Coltella L, Lucignano B, Pansani L, Ranno S, Russo C, Urbani A, Federici G, Menichella D.
SourceMicrobiology Unit, Children's Hospital and Research Institute Bambino Gesù, Piazza Sant'Onofrio 4, 00165, Rome, Italy. lorenza.putignani@opbg.net

Abstract
Proteomics is particularly suitable for characterising human pathogens with high life cycle complexity, such as fungi. Protein content and expression levels may be affected by growth states and life cycle morphs and correlate to species and strain variation. Identification and typing of fungi by conventional methods are often difficult, time-consuming and frequently, for unusual species, inconclusive. Proteomic phenotypes from MALDI-TOF MS were employed as analytical and typing expression profiling of yeast, yeast-like species and strain variants in order to achieve a microbial proteomics population study. Spectra from 303 clinical isolates were generated and processed by standard pattern matching with a MALDI-TOF Biotyper (MT). Identifications (IDs) were compared to a reference biochemical-based system (Vitek-2) and, when discordant, MT IDs were verified with genotyping IDs, obtained by sequencing the 25-28S rRNA hypervariable D2 region. Spectra were converted into virtual gel-like formats, and hierarchical clustering analysis was performed for 274 Candida profiles to investigate species and strain typing correlation. MT provided 257/303 IDs consistent with Vitek-2 ones. However, amongst 26/303 discordant MT IDs, only 5 appeared "true". No MT identification was achieved for 20/303 isolates for incompleteness of database species variants. Candida spectra clustering agreed with identified species and topology of Candida albicans and Candida parapsilosis specific dendrograms. MT IDs show a high analytical performance and profiling heterogeneity which seems to complement or even outclass existing typing tools. This variability reflects the high biological complexity of yeasts and may be properly exploited to provide epidemiological tracing and infection dispersion patterns.

细菌耐药

J Clin Microbiol. 2010 Mar;48(3):900-7. Epub 2010 Jan 6.
High-throughput identification of bacteria and yeast by matrix-assisted laser desorption ionization-time of flight mass spectrometry in conventional medical microbiology laboratories.
van Veen SQ, Claas EC, Kuijper EJ.
SourceDepartment of Medical Microbiology, Leiden University Medical Center, Leiden, Netherlands. s.q.van_veen@lumc.nl

Abstract
Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is suitable for high-throughput and rapid diagnostics at low costs and can be considered an alternative for conventional biochemical and molecular identification systems in a conventional microbiological laboratory. First, we evaluated MALDI-TOF MS using 327 clinical isolates previously cultured from patient materials and identified by conventional techniques (Vitek-II, API, and biochemical tests). Discrepancies were analyzed by molecular analysis of the 16S genes. Of 327 isolates, 95.1% were identified correctly to genus level, and 85.6% were identified to species level by MALDI-TOF MS. Second, we performed a prospective validation study, including 980 clinical isolates of bacteria and yeasts. Overall performance of MALDI-TOF MS was significantly better than conventional biochemical systems for correct species identification (92.2% and 83.1%, respectively) and produced fewer incorrect genus identifications (0.1% and 1.6%, respectively). Correct species identification by MALDI-TOF MS was observed in 97.7% of Enterobacteriaceae, 92% of nonfermentative Gram-negative bacteria, 94.3% of staphylococci, 84.8% of streptococci, 84% of a miscellaneous group (mainly Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella [HACEK]), and 85.2% of yeasts. MALDI-TOF MS had significantly better performance than conventional methods for species identification of staphylococci and genus identification of bacteria belonging to HACEK group. Misidentifications by MALDI-TOF MS were clearly associated with an absence of sufficient spectra from suitable reference strains in the MALDI-TOF MS database. We conclude that MALDI-TOF MS can be implemented easily for routine identification of bacteria (except for pneumococci and viridans streptococci) and yeasts in a medical microbiological laboratory.

附上文献的全文，供大家下载学习与讨论。

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二进制

资讯已阅，谢了！好仪器价码不菲，基层医院可望而不可及，唯羡慕份儿。

丁爱莲

东西确实好，好像在国内已有推出，能推广开吗？国内好像蛮重视药敏，对这个有需求吗？

桂花香

昨天我们讨论2012年抗菌药物专项整治方案的时候，领导还问微生物的组长，如果能够快速给临床提供微生物检验报告？能否在48小时内提供？组长说不行，如果有这样的仪器，就能实现快速检测的目的，真正为临床提供及时的服务。
只是不知道医院领导是否能够下决心购买？

veryoldman

跳跃式发展总是畸形的！设备的确不错，但细菌鉴定药敏之前的步骤如果得不到大的改观的话，招标买此类仪器就是浪费！

细菌耐药

细菌耐药 发表于 2012-2-18 07:08

                               
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J Clin Microbiol. 2010 Mar;48(3):900-7. Epub 2010 Jan 6.
High-throughput identification of bacteria ...

非常好的资料，谢谢分享

鬼才

细菌耐药 发表于 2012-4-4 11:15

                               
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非常好的资料，谢谢分享

现在有许多先进的技术在基层医院根本用不上，这也是我在工作中的困惑。从上月我返回岗位以来，一直在为如何把自己的科室带动起来，走向感控前沿而苦恼着，并非领导不支持，而是我们这里的确缺少人才、设备和资金。

ryanma

应该说，细菌的临床检测分为培养、鉴定和药敏三个阶段或时间段。这样的仪器设备的确是方法上的革命，可以减低细菌鉴定的时间，但对培养和药敏并未产生影响。估计只对部分目前难鉴定细菌有效。就鉴定一步来说，比现在的自动鉴定方法如化学法等能减低多少呢，对于临床出报告效率能产生多大的影响呢？尚难说。

婉若秋水

还是没有弄明白缩短工作时限的原理？在给其他人员讲解这个仪器时遭到质疑。
能否给一个通俗的解释？

细菌耐药

婉若秋水 发表于 2012-12-22 21:45

                               
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还是没有弄明白缩短工作时限的原理？在给其他人员讲解这个仪器时遭到质疑。
能否给一个通俗的解释 ...

简单地说，是根据细菌蛋白谱的差异来鉴定不同的细菌。