Safety and Bruker Emerging Technologies & MALDI-TOF. Tom Tague, Ph.D.
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1 Safety and Bruker Emerging Technologies & MALDI-TOF Tom Tague, Ph.D.
2 Bruker Today Global Corporation > $1.7 billion revenue p.a. (2017) Founded in 1960 in Karlsruhe (DE) by Prof. Günther Laukien, with Nuclear Magnetic Resonance products
3 Bruker Corporation Diagnostic, Life Science and Analytical Systems Bruker Corporation (NASDAQ: BRKR) is a global manufacturer of high-end Diagnostic, Life Science and Analytical instruments Bruker s product lines include: magnetic technologies, spectroscopes and spectrometers, atomic force microscopes, diagnostic products and X-ray technologies Bruker Daltonics is part of Bruker Corporation and specialized in innovative turnkey mass spectrometry systems for clinical, academic, pharmaceutical and food applications (> 10,000 Mass spectrometers installed worldwide) Bruker operates in the diagnostics & life science industry and has developed a strong brand for innovation, service and quality 3
4 Bruker Safety Program Voluntary participation Chair Tom Tague, Ph.D. Employees from various division/buildings Meet quarterly Conduct regular walk-throughs of Billerica facilities Address safety issues raised by employees Limited time commitment Do not have the resources to fulfill all safety requirements Can only make recommendations, no authority to mandate Leaves Bruker vulnerable for lack of regulatory compliance
5 Many Regulations Require Programs Programs Many require written programs similar to ISO program Hazard Assessments Training Annual (although some every 2-3 years) Retraining Whenever there are changes to tasks Diligent record keeping of training usually required Updated lists Equipment, chemicals, personnel, responsibilities Reports Accident reporting Corrective actions (like in ISO) Some may need to be submitted to agencies (EPA, OSHA)
6 OSHA Regulations Which Ones Might Apply to Bruker Safe work practices Chemical safety Cryogen safety Blood born pathogen etc. etc. etc.
7 OSHA Regs Requiring Programs Chemical Safety Emergency Action Plan ( ) Fire Prevention ( ) Storage of Flammable and Combustible Liquids ( ) Hazardous Waste Operations & Emergency Response ( ) Portable Fire Extinguishers ( ) Toxic and Hazardous Subtances ( ) Ionizing Radiation ( ) Hazard Communication ( ) Global Harmonization System labelling (new 2013)
8 OSHA Regs Requiring Programs at Bruker Medical Medical Services and First Aid ( ) Bloodborne Pathogens ( ) (Bruker BioSafety Labs are a great example of a compliance program) Injury & Illness Recordkeeping OSHA 300 and 301 logs
9 OSHA Regs Requiring Programs at Bruker Personal Protective Equipment Hearing ( ) Respiratory ( ) Eye and Face ( ) Head ( ) Foot ( ) Hand ( ) Ergonomics (Guidelines only) but enforced through General Duty clause Determined by Hazard Assessments
10 OSHA Regs Requiring Programs at Bruker Electrical / Mechanical Operations of Powered Platform ( ) Control of Hazardous Energy ( ) Powered Industrial Trucks ( ) Machine Guarding ( ) Electrical Safety-Related Work ( ) Electrical Protective Equipment ( ) NFPA 72 Arc-flash Protection
11 OSHA Regs Requiring Programs at Bruker Others Accident Prevention Signs and Tags ( ) Permit-Required Confined Space ( ) Welding ( ) Accident Investigation (General Duty Clause invoked) Preventing violence in the Workplace (???) Recommended Pre-inspection plan (just in case OSHA shows up)
12 OSHA Responsibilities at Bruker? Management Comply with Regulations and provide a Safe Workplace Create and communicate Policies and Procedures Regular updates and retraining required Reporting certain incidents promptly to OSHA Record and post OSHA-mandated info Form 300 and 301, citations, etc Investigate accidents & correct procedures and practices Enforce policies using employee disciplinary action Employees Following Policies and Procedures (i.e. SOPs) Audits employee compliance (performance review)
13 Example: A Program for Hazard Communication Written & updated regularly with every workplace change Jobs, tasks, equipment, responsibilities Training programs General - every year Task specific with each job description All new employees All visitors or contractors Record-keeping Requirements Chemicals Inventory SDSs (universally accessible to employees, visitors, & inspectors) Labels
14 Recommendations Clerical Voluminous record-keeping requirements Much is similar to record-keeping found in HR Compliance Management Oversees all compliance programs Best if trained in Occupational Safety or related experience Reports directly to upper management Has authority to require changes Responsible for program development & implementation Works with Line Managers and the Safety Committee Compliance audits Recommends changes
15 Benefits of Creating a Compliance Program Comply with Federal Regulations Avoid fines and liability of corporate management Improved ability to compete Government contracts often require compliance documentation Bruker often struggles to comply More customer contracts require safety program audits Other agencies (FDA, EPA, DEP) may require documentation of compliance to OSHA Reduce workplace injuries and illnesses i.e. a safer workplace Reduce workers compensation costs Reduce insurance costs Budget compliance activities
16 A Proactive Approach to Safety Passive past approach to Safety Obstacles to a proactive approach Change of culture! ES&H documentation likely lacking or outdated PPE compliance non-existent Completely unsafe cluttered and unsafe No Safety Officer Limited budget for training and PPE
17 A Proactive Approach to Safety Current proactive approach to Safety Training conducted across OSHA required items External auditor used to assess current state of workplace safety Training records tracked carefully Rewrite of ES&H manual (to be updated annually)
18 Tools Internet OSHA.gov DOT.gov EPA.gov MSHA.gov Safety Software
19 How Can Bruker Comply With the Regs? Safety Committee Volunteers with primary responsibilities elsewhere Management Not previously involved in the day-to-day compliance effort Managers Primary responsibility for procedures and task-specific training Informing managers of their responsibilities in a safe workplace Record-keeping This is the most time-consuming responsibility Training Equipment Inventories
20 How Does Bruker Comply With OSHA Regs? Resources - THE issue Safety Committee Can provide oversight or audit capabilities Tools software packages, manuals, regulations External Risk Management partner can be an inexpensive way to get on the right path Managers Specific Programs Electrical (facilities & service) Machine Guarding (machine shop) Record-keeping Who will be responsible for keeping records, accessible for inspection? Preparing reports/summaries with customers requiring our compliance
21 How Can Bruker Comply With the Regs? Management structure that includes regulatory compliance Support of HR is key Occupational Health and Safety or Compliance professional Clerical Staff or online partner like JJKeller to assist in compliance record-keeping
22 BSL rated lab and BBP safety How do you identify unknown potential biological hazards? Can be small quantities Can be almost any type
23 Bruker Corporation Bruker Microbiology & Diagnostics Product overview MALDI Biotyper For microbial identification and more Easy workflow Fast Time-To-Result Specific Robust Cost-effective Consumables and accessories for optimized microbial identification workflows Disposable MBT Biotargets 96 MBT Pilot workstation for guided sample transfer MBT Galaxy for automated application of matrix and formic acid Fast and direct identification of microorganisms from positive blood cultures with the MBT Sepsityper IVD Assay MBT STAR-BL IVD Assays for resistance detection via MALDI-TOF mass spectrometry Confident Mycobacteria identification
24 MALDI Biotyper Timeline 1960 Foundation of the Bruker company 1988 Establishment of the mass spectrometry business in Bremen, Germany 1991 Introduction of the first MALDI-TOF mass spectrometer 1998 Microbiological R&D group starts operation 2007 Establishment of ISO in addition to existing ISO First MALDI Biotyper systems in routine laboratories 2009 Launch of the MALDI Biotyper IVD system 2013 FDA Clearance of the MALDI Biotyper CA system 12/2017 AOAC-OMA approvals, OMA# & OMA# /2018 ISO part 6 validation by MicroVal 04/2018 Over 2,900 MALDI Biotyper systems sold
25 MALDI Biotyper Identify Gram+, Gram- bacteria, yeasts and fungi by their unique proteomic fingerprint Automated ID, Manual ID, Gram staining, Oxidase, Coagulase, Catalase, Strep grouping, Tributrine, Urea, Masterlex, Agglutinations, Specialised media, Morphology 1 min Bacillus subtilis Escherichia coli Candida albicans Aspergillus fumigatus
26 MALDI Biotyper Benefits Speed => Very fast identification of causative microorganisms Highly accurate identification results => Reliable Ease of use =>Low training efforts needed for new staff Lower cost per sample than conventional methods Regular library updates => Continuous expansion of identification performance Possibility to create own reference libraries => Flexible
27 MALDI Biotyper Complete solution for fast and easy microorganism identification MALDI-TOF System Sample preparation protocols Consumables Acquisition and data processing software Reference library Statistical tools Application Support Service
28 MALDI Biotyper Components How does it work
29 MALDI Biotyper System Core Components Dedicated MBT Software for Automated data acquisition Automated data processing Automated pattern matching Report creation MBT Reference Library 2,748 species entries Continuously maintained & updated MALDI Biotyper Spectrometer Benchtop instrument Unattended Operation Consumables Bruker HCCA Matrix Bruker Bacterial Test Standard MBT Biotarget 96
30 MALDI Biotyper Microorganism Identification by MALDI-TOF MS: Workflow 96 samples identified in ~60 minutes Identification Selection of colony of unknownresults microorganism comparable to sequencing Preparation onto MALDI target plate Faster than conventional techniques Cost-effective Identification by pattern matching Acquisition of MALDI-TOF profile spectrum
31 MALDI Biotyper - Basics Matrix Assisted Laser Desorption / Ionization Time Of Flight Mass Spectrometry
32 MALDI Biotyper - Basics Matrix Assisted Laser Desorption / Ionization Time Of Flight Mass Spectrometry
33 MALDI Biotyper - Basics Matrix Assisted Laser Desorption / Ionization Time Of Flight Mass Spectrometry Time-of-Flight E kin = mv 2 /2 m/z values Intensity m/z
34 MALDI Biotyper - Basics Robust identification method, as it relies on highly abundant proteins Intens. [a.u.] ribosomal Protein m/z RL ,33 RS ,82 RL ,39 RL33meth. 6255,39 RL ,19 RL ,60 RL ,74 RL ,45 RL ,06 RS , Mass range calibrated: m/z E. coli 2,000 20,000 Da
35 MALDI Biotyper Sample preparation procedures EASY and FAST Direct transfer (for 90-95% of microorganisms) - Direct transfer of biological material on MALDI Target - After all samples are processed, add MALDI Matrix - Usually perfect for gram-negative and most gram-positive microorganisms Extended direct transfer method (e.g. for yeast) - Direct transfer of biological material on MALDI Target - Add drop of formic acid ( on target extraction ) - Dry at room temperature - Add matrix Formic acid extraction (rare) - A series of steps are performed in an Eppendorf tube to extract proteins from isolated colonies Direct transfer using a tooth pick
36 MALDI Biotyper Sample preparation procedures - example 1.5 Candida krusei DT Candida krusei edt Candida krusei Ex m/z
37 MALDI Biotyper Run samples from your media of choice Psdm. oleovorans B396_Medium 360 Psdm. oleovorans B396_Medium 464 Psdm. oleovorans B396_Medium 53 Psdm. oleovorans B396_Medium 65 Psdm. oleovorans B396_Medium 98 Psdm. oleovorans B396_MRS10 Species ID does not depend on growth conditions, or medium selection, as illustrated by (e.g.): Psdm. oleovorans B396_YPD Veloo et al. Clinical Microbiology and Infection 2011 Lartigue et al. JCM 2009 Pseudomonas oleovorans Mellmann grown on et different al. JCM media m/z
38 MALDI Biotyper - Bioinformatics Unsupervised Main Spectra (MSP) concept Score base pattern matching Unknown microorganism is matched against each Main Spectrum in the library Calculation of a matching score based on: Matches MSP to unknown % matches of the reference spectrum (e.g. 6/10) Matches unknown to MSP % matches of the unknown spectrum (e.g. 6/20 = 3/10) Correlation of intensities value of intensity correlation
39 MALDI Biotyper Reference library Bruker MBT Compass Library (version April 2018) 2,748 different species MSPs have been created from industrial, veterinary and clinical isolates provided by collaborating partners, round robin strains and strains from isolates from accredited ISO 9001:2008 certified strain collections Every Microorganism could be of Relevance
40 Summary Safety regulations continue to become more important to understand and comply with Proactive approach to safety is a key to success in minimizing risk MALDI is an important tool for identifying biological risk
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