MALDI Biotyper CA (MBT-CA) System, MBT smart CA System

K163536 · Bruker Daltonik GmbH · PEX · Jul 26, 2017 · Microbiology

Device Facts

• Record ID: K163536
• Device Name: MALDI Biotyper CA (MBT-CA) System, MBT smart CA System
• Applicant: Bruker Daltonik GmbH
• Product Code: PEX · Microbiology
• Decision Date: Jul 26, 2017
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3361
• Device Class: Class 2

Intended Use

The MALDI Biotyper CA System is a mass spectrometer system using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) for the identification of microorganisms cultured from human specimens. The MALDI Biotyper CA System is a qualitative in vitro diagnostic device indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and yeast infections.

Device Story

The MBT-CA System uses MALDI-TOF mass spectrometry to identify microorganisms from cultured human specimens. Input: isolated microbial colonies from various agar media. Process: colonies are transferred to a target plate, overlaid with matrix (α-Cyano-4-hydroxycinnamic acid), and ionized via laser (nitrogen or smartbeam). The system measures the time-of-flight of ionized ribosomal proteins to generate a mass spectrum. Software compares this spectrum against a reference library using a biostatistical algorithm to produce a log(score) (0.00–3.00). Output: identification result with confidence level. Used in clinical laboratories by trained microbiologists. Results are interpreted alongside Gram stain, morphology, and clinical context to aid in diagnosing bacterial and yeast infections.

Clinical Evidence

Clinical method comparison study across four US sites and an in-house lab tested 2,091 fresh/stored isolates. Performance compared to reference sequencing (16S rRNA/ITS/protein gene). Overall high-confidence identification (log score ≥2.0) was 98.45% for high-resolution species. Reproducibility study (3 sites, 2 operators, 4 instruments) showed 99% correct identification. Challenge panel (46 organisms) confirmed workflow reliability. Additional analytical studies validated new media, target plates, and smartbeam laser technology, showing 0% false identification rates in validation cohorts.

Technological Characteristics

Mass spectrometer using MALDI-TOF principle. Materials: reusable steel targets (48-spot) or disposable targets (96-spot). Matrix: α-Cyano-4-hydroxycinnamic acid. Energy source: nitrogen laser or smartbeam laser. Connectivity: standalone system with integrated software. Software: biostatistical matching algorithm comparing spectra to reference library. Sterilization: N/A (laboratory instrument).

Indications for Use

Indicated for identification of cultured bacterial and yeast microorganisms from human specimens to aid in diagnosis of infections. For use by trained laboratory personnel.

Regulatory Classification

Identification

A mass spectrometer system for clinical use for the identification of microorganisms is a qualitative in vitro diagnostic device intended for the identification of microorganisms cultured from human specimens. The device is comprised of an ionization source, a mass analyzer, and a spectral database. The device is indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and fungal infections.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed documentation for device software, including, but not limited to, standalone software applications and hardware-based devices that incorporate software. (2) Premarket notification submissions must include database implementation methodology, construction parameters, and quality assurance protocols. (3) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling. (4) As part of the risk management activities performed under 21 CFR 820.10(c) design and development, you must document an appropriate end user device training program that will be offered as part of your efforts to mitigate the risk of failure to correctly operate the instrument. (5) Premarket notification submissions must include details on the appropriate end user device training program that will be offered while marketing the device.

Predicate Devices

• MBT-CA System (K142677)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo consists of a stylized eagle or bird symbol, composed of three overlapping profiles facing to the right. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the bird symbol. The logo is black and white.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

July 26, 2017

Bruker Daltonik GmbH Mr. David Cromwick Director of Quality 40 Manning Rd Billerica. MA 01821 US

Re: K163536

Trade/Device Name: MALDI Biotyper CA (MBT-CA) System, MBT smart CA System Regulation Number: 21 CFR 866.3361 Regulation Name: Mass spectrometer system for clinical use for the identification of microorganisms Regulatory Class: II Product Code: PEX Dated: December 16, 2016 Received: December 16, 2016

Dear Mr. Cromwick:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of

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medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809). please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours.

Kristian M. Roth-S

For: Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) K163536

Device Name MALDI Biotyper CA System

#### Indications for Use (Describe)

The MALDI Biotyper CA System is a mass spectrometer systems using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) for the identification of microorganisms cultured from human specimens.

The MALDI Biotyper CA System is a qualitative in vitro diagnostic device indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and yeast infections.

The following organisms are claimed:

Bacteria: Abiotrophia defectiva Achromobacter xylosoxidans Acinetobacter baumannii / nosocomialis group Acinetobacter calcoaceticus Acinetobacter haemolyticus Acinetobacter johnsonii Acinetobacter junii Acinetobacter lwoffii Acinetobacter pittii Acinetobacter radioresistens Acinetobacter ursingii Actinomyces europaeus Actinomyces funkei Actinomyces graevenitzii Actinomyces hyovaginalis Actinomyces meyeri Actinomyces neuii Actinomyces odontolyticus Actinomyces oris Actinomyces radingae Actinomyces turicensis Actinomyces urogenitalis Actinotignum schaalii group Aerococcus sanguinicola Aerococcus urinae Aerococcus viridans Aeromonas salmonicida Aeromonas sp[7] Aggregatibacter actinomycetemcomitans Aggregatibacter aphrophilus Aggregatibacter segnis Alcaligenes faecalis Alloiococcus otitis Alloscardovia omnicolens

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Anaerococcus murdochii Anaerococcus vaginalis Arthrobacter cumminsii Bacteroides caccae Bacteroides fragilis Bacteroides nordii Bacteroides ovatus group Bacteroides pyogenes Bacteroides salyersiae Bacteroides stercoris group Bacteroides thetaiotaomicron group Bacteroides uniformis Bacteroides vulgatus group Bifidobacterium breve Bordetella group[3] Bordetella hinzii Brevibacterium casei Brevundimonas diminuta group Burkholderia cepacia complex [13] Burkholderia gladioli Burkholderia multivorans Campylobacter coli Campylobacter jejuni Campylobacter ureolyticus Capnocytophaga ochracea Capnocytophaga sputigena Chryseobacterium gleum Chryseobacterium indologenes Citrobacter amalonaticus complex Citrobacter freundii complex Citrobacter koseri Clostridium beijerinckii Clostridium bifermentans Clostridium butyricum Clostridium clostridioforme group Clostridium difficile Clostridium innocuum Clostridium paraputrificum Clostridium perfringens Clostridium ramosum Clostridium septicum Clostridium sordellii Clostridium sporogenes / Clostridium botulinum (group I) Clostridium tertium Corynebacterium accolens Corynebacterium afermentans group Corynebacterium amycolatum Corynebacterium aurimucosum group Corynebacterium bovis Corynebacterium coyleae Corynebacterium diphtheriae Corynebacterium freneyi Corynebacterium glucuronolyticum Corynebacterium glutamicum

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Corynebacterium jeikeium Corynebacterium kroppenstedtii Corynebacterium macginleyi Corynebacterium minutissimum Corynebacterium mucifaciens / ureicelerivorans group Corynebacterium propinquum Corynebacterium pseudodiphtheriticum Corynebacterium pseudotuberculosis Corynebacterium resistens Corynebacterium riegelii Corynebacterium striatum group Corynebacterium tuberculostearicum Corynebacterium ulcerans Corynebacterium urealyticum Corynebacterium xerosis Cronobacter sakazakii group Cupriavidus pauculus group Delftia acidovorans group Dermabacter hominis Dermacoccus nishinomiyaensis Edwardsiella tarda Eikenella corrodens Elizabethkingia meningoseptica group Enterobacter aerogenes Enterobacter amnigenus Enterobacter cloacae complex Enterococcus avium Enterococcus casseliflavus Enterococcus durans Enterococcus faecalis Enterococcus faecium Enterococcus gallinarum Enterococcus hirae Enterococcus mundtii Enterococcus raffinosus Escherichia coli Escherichia hermannii Escherichia vulneris Ewingella americana Facklamia hominis Finegoldia magna Fluoribacter bozemanae Fusobacterium canifelinum Fusobacterium necrophorum Fusobacterium nucleatum Gardnerella vaginalis Gemella haemolysans Gemella morbillorum Gemella sanguinis Granulicatella adiacens Haemophilus haemolyticus Haemophilus influenzae Haemophilus parahaemolyticus group Haemophilus parainfluenzae

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Hafnia alvei Helcococcus kunzii Kingella denitrificans Kingella kingae Klebsiella oxytoca / Raoultella ornithinolytica Klebsiella pneumoniae Klebsiella variicola Kocuria kristinae Kytococcus sedentarius Lactobacillus gasseri Lactobacillus jensenii Lactobacillus rhamnosus Lactococcus garvieae Lactococcus lactis Leclercia adecarboxylata Legionella longbeachae Legionella pneumophila Leuconostoc citreum Leuconostoc mesenteroides Leuconostoc pseudomesenteroides Listeria monocytogenes Macrococcus caseolyticus Mannheimia haemolytica group Micrococcus luteus Micrococcus lylae Mobiluncus curtisii Moraxella sg Branhamella catarrhalis* Moraxella sg Moraxella nonliquefaciens* Moraxella sg Moraxella osloensis* Morganella morganii Myroides odoratimimus Myroides odoratus Neisseria bacilliformis Neisseria cinerea Neisseria elongata Neisseria flavescens / subflava group Neisseria gonorrhoeae Neisseria lactamica Neisseria meningitidis Neisseria sicca group Neisseria weaveri Nocardia brasiliensis Nocardia cyriacigeorgica Nocardia farcinica group Nocardia nova Nocardia otitidiscaviarum Ochrobactrum anthropi Oligella ureolytica Oligella urethralis Pantoea agglomerans Parabacteroides distasonis Parabacteroides goldsteinii Parabacteroides johnsonii / merdae group Parvimonas micra

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Pasteurella multocida Pediococcus acidilactici Pediococcus pentosaceus Peptoniphilus harei group Peptostreptococcus anaerobius Plesiomonas shigelloides Pluralibacter gergoviae Porphyromonas gingivalis Porphyromonas somerae Prevotella bivia Prevotella buccae Prevotella denticola Prevotella intermedia Prevotella melaninogenica Propionibacterium acnes Proteus mirabilis Proteus vulgaris group Providencia rettgeri Providencia stuartii Pseudomonas aeruginosa Pseudomonas fluorescens group Pseudomonas oryzihabitans Pseudomonas putida group Pseudomonas stutzeri Ralstonia pickettii Rhizobium radiobacter Rothia aeria Rothia dentocariosa Rothia mucilaginosa Salmonella sp** Serratia fonticola Serratia liquefaciens Serratia marcescens Serratia odorifera Serratia plymuthica Serratia rubidaea Sphingobacterium multivorum Sphingobacterium spiritivorum Sphingomonas paucimobilis group Staphylococcus aureus Staphylococcus auricularis Staphylococcus capitis Staphylococcus caprae Staphylococcus carnosus Staphylococcus cohnii Staphylococcus delphini Staphylococcus epidermidis Staphylococcus equorum Staphylococcus felis Staphylococcus haemolyticus Staphylococcus hominis Staphylococcus intermedius Staphylococcus lentus Staphylococcus lugdunensis

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Staphylococcus pasteuri Staphylococcus pettenkoferi Staphylococcus pseudintermedius Staphylococcus saccharolyticus Staphylococcus saprophyticus Staphylococcus schleiferi Staphylococcus sciuri Staphylococcus simulans Staphylococcus vitulinus Staphylococcus warneri Staphylococcus xylosus Stenotrophomonas maltophilia Streptococcus agalactiae Streptococcus anginosus Streptococcus canis Streptococcus constellatus Streptococcus dysgalactiae Streptococcus equi Streptococcus gallolyticus Streptococcus gordonii Streptococcus intermedius Streptococcus lutetiensis Streptococcus mitis / oralis group Streptococcus mutans Streptococcus parasanguinis Streptococcus pneumoniae Streptococcus pyogenes Streptococcus salivarius / vestibularis group Streptococcus sanguinis Streptococcus sobrinus Streptococcus thermophilus Sutterella wadsworthensis Trueperella bernardiae Turicella otitidis Vagococcus fluvialis Veillonella parvula group Vibrio parahaemolyticus Vibrio vulnificus Weeksella virosa Yersinia enterocolitica Yersinia frederiksenii Yersinia intermedia Yersinia kristensenii Yersinia pseudotuberculosis

*sg = subgenus **sp = species

Yeasts: Candida albicans Candida boidinii Candida dubliniensis Candida duobushaemulonii

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Type of Use (Select one or both, as applicable)

> Prescription Use (Part 21 CFR 801 Subpart D)

_ | Over-The-Counter Use (21 CFR 801 Subpart C)

#### CONTINUE ON A SEPARATE PAGE IF NEEDED.

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#### 10. 510(k) SUMMARY

| Date of Summary: | June 21, 2017 |
|----------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Product Name | MBT-CA System |
| Sponsor | Bruker Daltonik GmbH<br>Fahrenheitstrasse 4<br>28359 Bremen, Germany |
| Correspondent | Bruker Daltonik GmbH<br>Markus Kostrzewa, Vice President Clinical Mass Spectrometry R&D<br>Fahrenheitstrasse 4<br>28359 Bremen, Germany<br>Phone: [phone]<br>Fax: [phone]<br>Email: [e-mail] |
| Device Identification | |
| Trade or Proprietary Name: | MALDI Biotyper CA System |
| Common or Usual Name: | System, mass spectrometry, maldi tof, microorganism identification,<br>cultured isolates |
| Product Code: | PEX |
| Regulation Section: | 21 CFR 866.3361 Instrumentation for clinical multiplex test systems |
| Device Class: | Class II |
| Panel: | Microbiology |

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#### Substantial Equivalency

The MBT-CA System (K163536) is substantially equivalent to the MBT-CA System (K142677). Table 1 compares the characteristics of the MBT-CA System (New Device) and the MBT-CA System (Predicate Device).

| Similarities | | |
|------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | NEW DEVICE | PRIMARY PREDICATE DEVICE |
| Characteristic | MBT-CA System (K163536) | MBT-CA System (K142677) |
| Product Codes | PEX | PEX |
| Intended use | The MALDI Biotyper CA System is a mass<br>spectrometer systems using matrix-assisted<br>laser desorption/ionization - time of flight<br>(MALDI-TOF) for the identification of<br>microorganisms cultured from human<br>specimens.<br><br>The MALDI Biotyper CA System is a<br>qualitative in vitro diagnostic devices<br>indicated for use in conjunction with other<br>clinical and laboratory findings to aid in the<br>diagnosis of bacterial and yeast infections. | The MALDI Biotyper CA System is a mass<br>spectrometer system using matrix-assisted<br>laser desorption/ionization - time of flight<br>(MALDI-TOF) for the identification of<br>microorganisms cultured from human<br>specimens.<br><br>The MALDI Biotyper CA System is a<br>qualitative in vitro diagnostic device<br>indicated for use in conjunction with other<br>clinical and laboratory findings to aid in the<br>diagnosis of bacterial and yeast infections. |
| Sample type | Isolated colony from any patient sample<br>source.<br>Acceptable media:<br>• Columbia blood agar with 5% sheep<br>blood (Gram-negative bacteria)<br>• Trypticase soy agar with 5% sheep blood<br>(Gram-negative bacteria, Gram-positive<br>bacteria, yeasts)<br>• Chocolate agar (Gram-negative bacteria,<br>Gram-positive bacteria)<br>• MacConkey Agar (Gram-negative<br>bacteria)<br>• Columbia CNA agar with 5% sheep blood<br>(Gram-positive bacteria)<br>• Brucella Agar with 5% horse blood (Gram-<br>negative anaerobic bacteria, Gram-<br>positive anaerobic bacteria)<br>• CDC anaerobe Agar with 5% sheep blood<br>(Gram-negative anaerobic bacteria,<br>Gram-positive anaerobic bacteria)<br>• CDC anaerobe 5% sheep blood Agar with<br>phenylethyl alcohol (Gram-negative<br>anaerobic bacteria, Gram-positive<br>anaerobic bacteria)<br>• CDC anaerobe laked sheep blood Agar<br>with kanamycin and vancomycin (Gram-<br>negative anaerobic bacteria)<br>• Bacteroides bile esculin Agar with<br>amikacin ( <i>Bacteroides</i> species) | Isolated colony from any patient sample<br>source.<br>Acceptable media:<br>• Columbia blood agar with 5% sheep<br>blood (Gram-negative bacteria)<br>• Trypticase soy agar with 5% sheep blood<br>(Gram-negative bacteria, Gram-positive<br>bacteria, yeasts)<br>• Chocolate agar (Gram-negative bacteria,<br>Gram-positive bacteria)<br>• MacConkey Agar (Gram-negative<br>bacteria)<br>• Columbia CNA agar with 5% sheep blood<br>(Gram-positive bacteria)<br>• Brucella Agar with 5% horse blood (Gram-<br>negative anaerobic bacteria, Gram-<br>positive anaerobic bacteria)<br>• CDC anaerobe Agar with 5% sheep blood<br>(Gram-negative anaerobic bacteria,<br>Gram-positive anaerobic bacteria)<br>• CDC anaerobe 5% sheep blood Agar with<br>phenylethyl alcohol (Gram-negative<br>anaerobic bacteria, Gram-positive<br>anaerobic bacteria)<br>• CDC anaerobe laked sheep blood Agar<br>with kanamycin and vancomycin (Gram-<br>negative anaerobic bacteria)<br>• Bacteroides bile esculin Agar with<br>amikacin ( <i>Bacteroides</i> species) |
| Similarities | | |
| | NEW DEVICE<br>MBT-CA System (K163536) | PRIMARY PREDICATE DEVICE<br>MBT-CA System (K142677) |
| Characteristic | blood ( <i>Clostridium difficile</i> )<br>• Sabouraud-Dextrose Agar (Yeasts)<br>• Brain Heart Infusion Agar (Yeasts)<br>• <i>Campylobacter</i> Agar with 5 Antimicrobics<br>and 10% Sheep Blood ( <i>Campylobacter</i><br>species)<br>• Bordet Gengou Agar with 15% sheep<br>blood ( <i>Bordetella</i> species)<br>• plus three (3) additionally validated<br>media (see differences). | blood ( <i>Clostridium difficile</i> )<br>• Sabouraud-Dextrose Agar (Yeasts)<br>• Brain Heart Infusion Agar (Yeasts)<br>• <i>Campylobacter</i> Agar with 5 Antimicrobics<br>and 10% Sheep Blood ( <i>Campylobacter</i><br>species)<br>• Bordet Gengou Agar with 15% sheep<br>blood ( <i>Bordetella</i> species) |
| Type of Test | Automated Mass Spectrometry System | Automated Mass Spectrometry System |
| Matrix | α-Cyano-4-hydroxycinnamic acid | α-Cyano-4-hydroxycinnamic acid |
| Method of<br>Testing | Bacteria & Yeast: Direct testing<br><br>If after initial analysis the log(score) is<br>reported at <2.00, organisms may be<br>processed using the Extraction (Ext)<br>procedure or extended Direct Transfer (eDT,<br>70% aqueous formic acid) procedure. If eDT<br>procedure still yields log(score) <2.00,<br>organisms may be processed via Ext<br>procedure. | Bacteria & Yeast: Direct testing<br><br>If after initial analysis the log(score) is<br>reported at <2.00, organisms may be<br>processed using the Extraction (Ext)<br>procedure or extended Direct Transfer (eDT,<br>70% aqueous formic acid) procedure. If eDT<br>method still yields log(score) <2.00,<br>organisms may be processed via Ext<br>procedure. |
| Result Reporting | Organism identification is reported with<br>high confidence if the log(score) is ≥2.00.<br><br>An organism identification is reported with<br>low confidence if the log(score) is between<br>1.70 and <2.00. | Organism identification is reported with<br>high confidence if the log(score) is ≥2.00.<br><br>An organism identification is reported with<br>low confidence if the log(score) is between<br>1.70 and <2.00. |
| Matching<br>Algorithm | Calculates matches by comparing a new<br>spectrum against each single reference<br>entry of a reference database. | Calculates matches by comparing a new<br>spectrum against each single reference<br>entry of a reference database. |
| Recorded mass<br>range | 2,000 - 20,000 m/z | 2,000 - 20,000 m/z |
| Calibration | Bruker US IVD Bacterial Test Standard (BTS) | Bruker US IVD Bacterial Test Standard (BTS) |
| Database | MALDI Biotyper Reference Library for<br>Clinical Applications (MBT-CA) - update | MALDI Biotyper Reference Library for<br>Clinical Applications (MBT-CA) |

## Table 1: Substantial Equivalency Table

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| Differences | | |
|--------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------|
| Characteristic | NEW DEVICE<br>MBT-CA System (K163536) | PRIMARY PREDICATE DEVICE<br>MBT-CA System (K142677) |
| System Update | System claims additional organisms but no additional changes. | N/A |
| Media | Additional Media validated:<br>• Buffered Charcoal Yeast Extract Agar ( <i>Legionella</i> species)<br>• Buffered Charcoal Yeast Extract Selective Agar with polymyxin, anisomycin and vancomycin ( <i>Nocardia</i> species)<br>• Modified Thayer-Martin Agar ( <i>Neisseria</i> species) | See Similarities |
| MALDI-TOF MS instruments | Bruker microflex LT/SH (benchtop)<br>Bruker microflex LT/SH smart (benchtop) | Bruker microflex LT/SH (benchtop) |
| MALDI Target Plates | US IVD 48 Spot Target (48 positions reusable steel targets)<br>MBT Biotarget 96 US IVD (96 positions disposable targets) | US IVD 48 Spot Target (48 positions reusable steel targets) |

The differences do not affect substantial equivalence of the MBT-CA System and MBT-CA System (K142677). Both systems are mass spectrometers using matrix-assisted laser desorption/ionization-time to flight (MALDI-TOF) for the identification of microorganisms cultured from human specimens. The differences noted above do not impact the intended use and do not raise questions as to the safety and effectiveness of the test (new) device.

#### Intended Use

The MALDI Biotyper CA System is a mass spectrometer system using matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) for the identification of microorganisms cultured from human specimens.

The MALDI Biotyper CA System is a qualitative in vitro diagnostic device indicated for use in conjunction with other clinical and laboratory findings to aid in the diagnosis of bacterial and veast infections.

#### Methodology

Biochemical methods are currently the most commonly used methods for the identification of microorganisms. Organisms are tested against a range of reagents and organism identification is based on a microorganism's reaction to these reagents.

The MBT-CA System uses a different methodology for organism identification based on unique protein patterns of the microorganisms obtained from mass spectrometry. The test organism's spectrum (a pattern of mass peaks) is compared with a reference spectra library (database). Using biostatistical analysis, a probability ranking of the organism identification is generated. The probability ranking is represented as a log(score) between 0.00 and 3.00. Organism identification is reported with high confidence if the log(score) is ≥2.00. An organism identification is reported with low confidence if the log(score) is between 1.70 and <2.00.

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Organisms to be identified with the MBT-CA System should be isolated for purity on appropriate isolation media.

Direct Transfer procedure (DT): An individual colony from a subculture plate is transferred to a selected position on a US IVD 48 Spot Target plate or a MBT Biotarget 96 US IVD Target plate (targets) and overlaid with US IVD HCCA portioned (matrix). The standard solvent (50% acetonitrile / 47.5% H₂O / 2.5% trifluoroacetic acid) in the matrix solution extracts proteins (mainly ribosomal proteins, which are present in high concentration) from the microorganisms. When dried matrix crystallizes, the inoculated target is ready to be analyzed on the MBT-CA System. If after initial analysis the log(score) is reported as <2.00, organisms can be processed using the extended Direct Transfer (eDT) procedure or the Extraction procedure and analysis repeated. If eDT is employed and log(score) is reported as <2.00, reanalysis via the Extraction procedure may be used.

## extended Direct Transfer procedure (eDT):

If DT analysis yields a [log(score) <2.00] result, an individual colony from a subculture plate may be transferred to a selected position on a target and overlaid with 70% aqueous formic acid solution. The target is air-dried and then matrix is overlaid. When dried matrix crystallizes, the inoculated target is ready to be analyzed on the MBT-CA System. If a high confidence result is not achieved [log(score) is reported at <2.00], organisms can be processed using the Extraction procedure and analysis repeated.

Extraction procedure (Ext): If after initial analysis and eDT procedure the log(score) is reported at <2.00, organisms are processed using the Extraction procedure and analysis repeated. For this purpose, isolated colonies from the subculture plate are extracted in accordance with instructions of the user manual (Ext sample preparation procedure). An aliquot of extracted material is transferred to a selected position on a target, air-dried and then overlaid with matrix material. When dried matrix crystallizes, the inoculated target is ready to be analyzed on the MBT-CA System.

## MALDI-TOF Analysis:

Samples are analyzed using MALDI (matrix-assisted laser desorption/ionization) TOF (timeof-flight) mass spectrometry. The matrix transfers protons onto the extracted proteins and absorbs UV light. A laser in the MALDI- TOF mass spectrometer irradiates the matrix sample composite, causing evaporation and release of positively charged intact proteins and peptides ("soft" ionization technique). These ions are electrostatically accelerated over a short distance and arrive in the flight tube at a mass-dependent speed. As different proteins/peptides have different masses, ions arrive at the detector at different times (time of flight). The system measures the time (in the nanosecond range) between pulsed acceleration and the corresponding detector signal, the speed is converted into an exact molecular mass. The mass-to-charge ratio of an ion is proportional to the square of its drift time.

Highly abundant microbial proteins (mainly ribosomal proteins) result in a mass spectrum with characteristic mass and intensity distribution. It is species-specific for many bacteria and is interpreted as a molecular fingerprint to identify the test organism.

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Data acquisition is controlled with MBT-CA System software. The spectrum of the unknown organism is first transformed into a peak list. Using a biostatistical algorithm, this peak list is compared to the reference peak lists of organisms in the reference library (database) and a log(score) is generated. A higher log(score) indicates a higher degree of similarity to the organism in the reference library. Organism identification is reported with high confidence if the log(score) is ≥2.00. An organism identification is reported with low confidence if the log(score) is between 1.70 and <2.00.

The log(score) ranges, defined in the MBT-CA System, are indicative of the probability of organism identification. Results should be reviewed by a trained microbiologist and final organism identification should be based on all relevant information available. This information includes but is not limited to: Gram staining, colony morphology, growth characteristics, sample matrix, etc.

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#### Performance Data

#### Media and Colony Stability

With the inclusion of further Gram-negative and Gram-positive microaerophilic, additional Gram-negative and Gram-positive anaerobic and further Gram-negative and Gram-positive aerobic bacteria and yeasts, a study on the following media was conducted to confirm acceptability of the recommended agar and the stability of the colony for up to 12 hours storage at room temperature prior to analysis:

- Trypticase Soy Agar with 5% sheep blood (TSA)
- Buffered Charcoal Yeast Extract Agar (BCYE)
- . Buffered Charcoal Yeast Extract Selective Agar with polymyxin, anisomycin and vancomycin (BCYE/PAV)
- Modified Thayer-Martin Agar (MTM)

In the previousl submission (K142677) TSA medium was only validated for Gram-positive /negative aerobic bacteria and yeast. Goal for TSA medium in this submission was to show the suitability of this medium for Gram-positive /-negative microaerophilic and anaerobic bacteria.

Testing was conducted using one (1) Gram-negative bacterium, one (1) microaerophilic Gram-negative bacterium, one (1) Gram-positive bacterium, one (1) microaerophilic Grampositive bacterium, two (2) Legionella species, three (3) Nocardia species and three (3) Neisseria species, at varying incubation time points in replicates of eight (8). Additionally, thirteen (13) clinical isolates derived from Neisseria species were tested in duplicate at varying incubation time points.

Conclusion Media and Colony Stability: Three (3) additional media could be validated successfully. TSA medium could be validated for all kinds of organisms. Cultivation of Neisseria on Modified Thayer-Martin Agar (MTM) should not be longer than 24 hours plus 12 hours storage at room temperature.

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#### Resolution of the Acinetobacter baumannii complex

This study served to evaluate an improved species resolution power of the MBT-CA System of the Acinetobacter baumannii complex (members: A. baumannii, A. calcoaceticus, A. nosocomialis, A. pittii) down to the species level after final extraction procedure. Sixty-six (66) clinical isolates of Acinetobacter baumannii complex collected during initial Clinical Method Comparison protocol (see K130831) and identified by 16S rRNA and/or protein gene sequencing were re-tested using Direct Transfer, extended Direct Transfer and Extraction procedure in parallel.

The study results showed that the Acinetobacter baumannii complex cannot be completely but partially resolved applying MBT-CA System workflow. The reference library/software will be updated to complex Acinetobacter baumannii and Acinetobacter nosocomialis under the Acinetobater baumannii/nosocomialis group. After formation of an Acinetobacter baumannii/nosocomialis group unambiguous MBT-CA identification with high confidence [log(score) ≥2.00] of Acinetobacter baumannii/nosocomialis group, Acinetobacter calcoaceticus and Acinetobacter pittii is possible if final Extraction procedure has been applied. A matching hint will be included in the system package insert which contains the following alert: The displayed species should be considered a member of the Acinetobacter baumannii complex. For organisms identified by the MBT-CA System as Acinetobacter calcoaceticus, Acinetobacter pittii or Acinetobacter baumannii/nosocomialis group the full Extraction procedure (Ext) is mandatory for secure species differentiation.

#### Cultivation of yeast organisms at 37 °C

This study served to show the general applicability of the MBT-CA System workflow for identification of yeasts cultivated at 37 (± 2)°C. Testing was conducted using three (3) yeast species at varying incubation time points in replicates of eight (8). After initial testing, isolates were further tested at room temperature after twelve (12) hours post-incubation.

The study results confirmed the acceptability of 37 (± 2)°C cultivation of yeasts and sample colony stability of up to 12 hours.

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#### Reproducibility:

The reproducibility study for Gram-negative and Gram-positive aerobic bacteria, Gramnegative and Gram-positive microaerophilic bacteria, Gram-negative and Gram-positive anaerobic bacteria and yeasts was carried out to confirm day-to-day reproducibility and precision of the MALDI Biotyper CA System at different clinical study sites. The study was conducted for five (5) days with two (2) runs [two (2) operators] each day per clinical site. The sources of variability tested were:

- Two (2) operators/each clinical study site
- Three (3) clinical study sites
- At least two (2) target plates/each clinical study site
- Four (4) microflex LT/SH instruments

Ten (10) well-characterized organisms were chosen for this study and tested in duplicate via Direct Transfer procedure in accordance with product instructions. When the DT log(score) was <2.00, per product instructions, the test organism was tested following the extended Direct Transfer and Extraction procedure.

Note: As REPRO-2 was identified as Listeria monocytogenes after applying DT procedure test sites should have proceeded with testing of eDT and Ext procedure in parallel in accordance with the study protocol. Test sites did not conduct additional testing, therefore REPRO-02 was excluded from data analysis.

| Blinded Test Organism | Reproducibility<br>Panel | Site A:<br>MBT-CA ID<br>(DT+eDT+Ext) | Site B:<br>MBT-CA ID<br>(DT+eDT+Ext) | Site C:<br>MBT-CA ID<br>(DT+eDT+Ext) |
|--------------------------|--------------------------|--------------------------------------|--------------------------------------|--------------------------------------|
| Dermabacter hominis | REPRO-01 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Listeria monocytogenes | REPRO-02* | N/A | N/A | N/A |
| Nocardia farcinica group | REPRO-03 | 19/20 (95%) | 19/20 (95%) | 19/20 (95%) |
| Legionella pneumophila | REPRO-04 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Clostridium tertium | REPRO-05 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Facklamia hominis | REPRO-06 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Bacteroides caccae | REPRO-07 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Trueperella bernardiae | REPRO-08 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Neisseria meningitidis | REPRO-09 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| Rhodotorula mucilaginosa | REPRO-10 | 20/20 (100%) | 20/20 (100%) | 20/20 (100%) |
| TOTAL | | 179/180 (99%) | 179/180 (99%) | 179/180 (99%) |

Table 2: Reproducibility Study Summary

*REPRO-02: Organism skipped as MBT-CA ID applying DT procedure was not confirmed via eDT and Ext procedure.

Conclusion Reproducibility: 99% of test organisms were correctly identified with a log(score) ≥2.00 at each clinical test site. In addition, no isolates were falsely identified. Thus, data confirm reproducibility and precision of the whole MALDI Biotyper CA System independent from:

- Clinical Site
- System operators ●
- microflex LT/SH instruments
- Target plate

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#### Challenge Panel:

A panel of 46 organisms (12 Gram-positive aerobic bacteria, 8 Gram-negative aerobic bacteria, 4 Gram-positive microaerophilic bacteria, 5 Gram-negative microaerophilic bacteria, 10 Gram-positive anaerobic bacteria, 4 Gram-negative anaerobic bacteria, 3 yeasts) was tested at three (3) study sites. All of the forty-six (46) organisms included in the panel were selected from stored organisms tested during the clinical study. The study interim reference laboratory prepared the panel. Organism identifications were blinded to test sites. Each site tested the challenge panel member via Direct Transfer, extended Direct Transfer and Extraction procedure in parallel.

| Test procedure | Site A* | Site B** | Site C*** |
|-----------------|--------------|-------------|-------------|
| DT method | 43/45 (96%) | 38/44 (86%) | 36/41 (88%) |
| eDT method | 44/45 (98%) | 36/44 (82%) | 38/41 (93%) |
| Ext method | 43/45 (96%) | 36/44 (82%) | 39/41 (95%) |
| MBT-CA workflow | 45/45 (100%) | 40/44 (91%) | 40/41 (98%) |

#### Table 3: Challenge Panel Study Summary

* One (1) sample was not identified due to isolate failure to grow.

** Two (2) samples were not identified due to isolate failure to grow.

*** Five (5) samples were not identified due to isolate failure to grow.

#### Conclusion Challenge Panel:

Testing of the challenge panel confirms reliability of the MBT-CA System workflow independently from

- Clinical Site
- Test-operator ●
- microflex LT/SH mass spectrometer system
- US IVD 48 Spot Target plate

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Biological / Technical Equivalency Study - MBT-CA smart System

Objective of this validation was to demonstrate equivalence of the MBT-CA output when using MALDI-TOF mass spectrometers equipped with smartbeam laser technology and nitrogen laser technology.

A panel of thirty four (34) species which are part of the MBT-CA library were measured. The 34 species represent Gram negative, Gram positive, and yeast. All three sample preparation techniques (DT, eDT, Ext) were used in parallel (eDT and Ext were always additionally performed independent from the DT result). Each sample preparation technique (DT, eDT, Ext) was spotted 8 times onto the MALDI target. The measurement was performed on two nitrogen laser instruments and 3 smart laser instruments.

Overall 4080 spectra were represented in this study. The performance of all spectra for each single species, each single sample prep (DT, eDT and Ext), each MBT-CA result (no ID "red", low confidence ID "yellow", high confidence ID "green") for each instrument type is summarised in Table 4.

Table 4: Overall performance of 4080 measured samples. "High confidence ID" (green), "low confidence ID" (yellow) and "no ID" (ref) are shown. The table (left) shows the total numbers and table (right) the percentage.

| | Nitrogen (544) | | | Smart (816) | | | | Nitrogen (544) | | | Smart (816) | | |
|-----|----------------|-----|------|-------------|-----|------|-----|----------------|-------|-------|-------------|-------|-------|
| DT | 187 | 72 | 285 | 300 | 117 | 399 | DT | 34.4% | 13.2% | 52.4% | 36.8% | 14.3% | 48.9% |
| eDT | 37 | 61 | 446 | 63 | 120 | 633 | eDT | 6.8% | 11.2% | 82.0% | 7.7% | 14.7% | 77.6% |
| Ext | 2 | 3 | 539 | 0 | 13 | 803 | Ext | 0.4% | 0.6% | 99.1% | 0.0% | 1.6% | 98.4% |
| Σ | 226 | 136 | 1270 | 363 | 250 | 1835 | Σ | 13.8% | 8.3% | 77.8% | 14.8% | 10.2% | 75.0% |

As an example, the row "Ext" shows a similar percentage of "high confidence results" between both systems ( 99.1% for Nitrogen Laser versus 98.4% for Smartbeam Laser).

Conclusion: Based on these results the equivalence between the Nitrogen Laser System and the Smartbeam Laser System could be shown.

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Equivalence Study MBT Biotarget 96 US IVD (Nitrogen Laser)

This study was designed to verify and validate the use of the MBT Biotarget 96 US IVD in conjunction with the MBT-CA System (Nitrogen Laser). Study goal was to show the equivalency between the already cleared US IVD MSP 48 Target Polished Steel and the new MBT Biotarget 96 US IVD. The functionality and performance of the MBT-CA System using the MBT Biotarget 96 US IVD under varying conditions was shown. Test runs were performed always in parallel using US IVD MSP 48 Target Polished Steel plates.

The following six studies were performed:

- Repeatability / Precision
- Tolerance range (dynamic range) - limit of detection (LOD)
- Sample stability prior to matrix application
- Sample stability post matrix application
- Mass Accuracy / Target Edge Effects
- Identification of 50 FDA cleared organisms according to MBT-CA System workflow.

#### Repeatability and Precision

In this study, approximately one thousand (1000) measurements were carried out. In total, seven hundred and twenty (720) MBT-CA System identifications (10 species, triplicates, two target plates, 1 operator per day, 2 runs per day, 6 days) were performed. 100% of test organisms were correctly MBT-CA identified at high confidence level [log(score) ≥2.0] applying the MBT-CA System workflow (combination of DT, eDT and Ext procedure). The rate of MBT-CA System false identification was 0% for all test samples after final sample preparation procedure.

Both target types performed similarly.

## Tolerance Range (Dynamic Range) - Limit of Detection (LOD)

In this study, approximately one thousand five hundred (1500) measurements were performed. 10 different species were analysed representing Gram positive, Gram negative bacteria and yeast. All sample preparation techniques (DT, eDT and Ext) were conducted. Up to 5 dilutions steps were performed for each scenario. The rate of MBT-CA System false identifications at least at low confidence level [log(score) ≥1.70] was 0%. The estimated dynamic range (limit of detection) of bacteria and yeast onto the MBT Biotarget 96 US IVD / US IVD MSP 48 Target Polished Steel required for MBT-CA System identification did not show significant differences.

As a common limit of detection the amount of about 5*10 cells (or cell equivalents using extraction) which must be placed on a target spot could be defined. Since a sharp boundary is hard to define, some exceptions occurred during this study. But these exceptions occurred for different species, different sample preparation techniques and different target types. Therefore there is no general trend for a better limit of detection for one kind of target.

Therefore the equivalence between the MBT Biotarget 96 US IVD / US IVD MSP 48 Target Polished Steel could be shown.

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#### Sample Stability prior to Matrix application

In this study, approximately two thousand five hundred (2500) measurements were carried out. Five species were analyzed on two instruments. Measurement was applied directly after matrix application and after 15, 30, 60 and 120 minutes. All sample preparation techniques (DT, eDT and Ext) were conducted. Each sample was spotted and measured 8 times. The rate of MBT-CA System false identifications was 0% for all tests.

The performance was similar for both target types.

#### Sample Stability post Matrix application

In this study, approximately three thousand (3000) measurements were performed. Two temperature conditions were analysed in this study (21℃ and 25℃). All measurements were conducted using 5 different species on two MALDI instruments. All sample preparation techniques (DT, eDT and Ext) were performed and each sample was spotted 8 times on both target types. The ready prepared sample stability was analyzed immediately after matrix application (standard condition) and after 4h, 8h and 24h of aging.

No MBT-CA false identifications occurred. The performance of the MBT Biotarget 96 US IVD as compared to that of the US IVD MSP 48 Target Polished Steel could be shown.

#### Validation Study - Validation of 50 Representative Claimed Species

Approximately three hundred (300) measurements were carried out. In this study 50 claimed species representing Gram nega…