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.

Device Description

The MBT-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 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.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study details for the MALDI Biotyper CA (MBT-CA) System, based on the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The provided document does not explicitly state pre-defined acceptance criteria in terms of numerical thresholds for sensitivity, specificity, or overall accuracy. Instead, it presents performance results from various studies (reproducibility, challenge panel, method comparison) and concludes that the device performs acceptably.

However, based on the reported performance in the Method Comparison study, common metrics for identification systems would be:

Performance Metric (Interpreted)	Acceptance Criteria (Implied / Expected)	Reported Device Performance (Overall Isolate Performance from Table 6)
High Confidence ID Rate (≥ 2.0 log(score))	High, ideally >95% for species identification	1904 / 1930 = 98.65% (for high resolution species) (1904 + 130) / (1930 + 136) = 98.42% (for high & low resolution species/genus)
Low Confidence ID Rate (≥ 1.7 to < 2.0 log(score))	Low, indicating need for further action	23 / 1930 = 1.19% (for high resolution species) 5 / 136 = 3.68% (for low resolution species/genus)
Incorrect MBT-CA ID (≥ 1.7 log(score)) or No ID (< 1.7 log(score))	Very Low, ideally <1%	3 / 1930 = 0.16% (Incorrect for high resolution species) 1 / 136 = 0.74% (Incorrect for low resolution species/genus) Equivalent to 4 / 2066 = 0.19% (Total incorrect/no ID for positive cases)
Overall Identification Rate (High or Low Confidence)	High, ideally >95%	(1904+23) / 1930 = 99.84% (for high resolution species) (1904+130+23+5) / (1930+136) = 99.81% (for high & low resolution species/genus)
False Identification Rate	0% (critical for diagnostic accuracy)	0% reported across several validation studies (Repeatability/Precision, LOD, Sample Stability, Validation of 50 Representative Claimed Species, Nocardia Study). For the overall isolate performance, the "Incorrect MBT-CA ID" for positive cases (3+1=4) indicates a very low rate of incorrect IDs, which are distinct from "negative" cases. The document states "no isolates were falsely identified" in the reproducibility study and similar conclusions in other studies. For the method comparison, it is reported as 0% for negative cases and very low for positive cases.

Note: The "acceptance criteria" presented above are inferred from the strong performance and conclusions drawn in the document, rather than explicitly stated numerical targets prior to testing.

2. Sample Size Used for the Test Set and Data Provenance

Method Comparison Study (Overall Isolate Performance):
- Sample Size: 2091 fresh and stored organisms.
- Data Provenance: Organisms were tested at four (4) US clinical test sites and an in-house laboratory. Isolates were sub-cultured and sent to an interim reference laboratory and then to a sequencing reference laboratory for ground truth determination. This indicates prospective and retrospective data collection with a US origin.
Reproducibility Study:
- Sample Size: 9 unique organisms (REPRO-02 excluded). Each organism tested in duplicate, 5 days, 2 runs/day, 3 sites (9 organisms x 2 replicates x 5 days x 2 runs x 3 sites = 540 measurements). Total MBT-CA IDs for summary = 179/180 per site.
- Data Provenance: Conducted at three (3) clinical study sites (US, likely, given the FDA submission context). The organisms were "well-characterized," suggesting they might be reference strains or previously identified clinical isolates.
Challenge Panel Study:
- Sample Size: 46 organisms.
- Data Provenance: Selected from stored organisms from the clinical study, prepared by the interim reference laboratory. Tested at three (3) study sites (US, likely).
Biological/Technical Equivalency Studies:
- Sample Size: 34 species for laser equivalency (4080 spectra). Multiple species for target equivalency (e.g., 1000 measurements for repeatability/precision, 1500 for LOD, 2500 for sample stability prior to matrix, 3000 for post-matrix stability, 50 FDA cleared organisms, 1500 for mass accuracy/edge effects). Nocardia Study: 30 strains covering 6 species, resulting in ~15,000 measurements.
- Data Provenance: Not explicitly stated for specific origin, but these are technical validation studies performed by the manufacturer, likely controlled lab settings.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications

Method Comparison Study:
- The ground truth was established by sequencing (16S rRNA or ITS sequencing and protein gene sequencing). This relies on established molecular biology techniques, not human expert interpretation. While experts run and interpret these sequences, the core ground truth is the genetic information itself. The document does not specify a number or qualification of "experts" in the sense of clinical reviewers for ground truth determination but implies reliance on the robust and objective results of gene sequencing performed by a sequencing reference laboratory.
Reproducibility Study:
- Organisms were "well-characterized." The ground truth was presumably established by prior definitive identification methods, likely including gene sequencing or reputable reference lab methods. No mention of independent experts for this study's ground truth.
Challenge Panel Study:
- Organism identifications were "blinded to test sites," and the panel was prepared by the study interim reference laboratory. The ground truth was established by the reference lab, again likely through gold standard methods like sequencing.

4. Adjudication Method for the Test Set

The document does not describe an explicit adjudication method involving multiple human readers or a specific consensus process for discrepancies in the generated log(scores) or identifications against a human-read ground truth. Instead:

The ground truth for the organism identity itself (reference algorithm) was established by molecular sequencing.
The device's log(score) provides a quantitative measure of confidence. If the log(score) is too low (<1.70), the result is "no ID." If it's between 1.70 and <2.00, it's "low confidence."
For discrepancies in the device's workflow (e.g., if Direct Transfer yields a low score), the protocol specifies progressive steps (extended Direct Transfer, then Extraction procedure) until a high confidence log(score) is achieved or the organism remains unidentified. This is an internal algorithmic "adjudication" within the device's workflow.
The final interpretation is stated to be by a "trained microbiologist," who should consider "all relevant information available" (Gram staining, colony morphology, growth characteristics, sample matrix, etc.) in conjunction with the device's results. This implies human oversight and potential adjudication of the device's output in a clinical setting, but not as part of the ground truth establishment for the studies described.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No MRMC comparative effectiveness study is described where human readers' performance with and without AI (MALDI-TOF) assistance is measured to determine an effect size. The studies focus on the standalone performance of the MALDI Biotyper CA System against a molecularly derived ground truth. The device is intended to aid in diagnosis, implying it would be used by humans, but the studies do not quantify the improvement in human performance with the device.

6. Standalone Performance Study (Algorithm Only)

Yes, extensive standalone performance studies were done. The "Method Comparison" study, "Reproducibility" study, and "Challenge Panel" study, as well as the various "Equivalency Studies," all evaluate the MALDI Biotyper CA System's performance (algorithm only, with human operation but without human interpretation as the ground truth) against a molecularly derived ground truth or against itself under different conditions. The tables showing "Overall Isolate Performance" (Table 6, 7, 8) are prime examples of standalone performance evaluation.

7. Type of Ground Truth Used

Method Comparison and Challenge Panel Studies: The primary ground truth for organism identification was established by molecular sequencing, specifically 16S rRNA or ITS sequencing and protein gene sequencing.
Reproducibility Study: Organisms were "well-characterized," implying a gold standard identification method, most likely molecular sequencing.
Equivalency Studies: These compared the performance of different components of the MBT-CA system (e.g., laser types, target plates) against each other or against established performance metrics, rather than identifying unknown organisms from scratch using an external ground truth. The underlying ground truth for the organisms used in these studies would still be molecular identification.

8. Sample Size for the Training Set

The document does not explicitly state the sample size used for training the MBT-CA System's reference database. It mentions the "MALDI Biotyper Reference Library for Clinical Applications (MBT-CA)" and states it's an "update" (K163536 vs. K142677). This reference library is what the device compares new spectra against. The training set would be the collection of reference spectra used to build this database. The provided text refers to new organisms added to the library but does not give a total size or the methodology used to build the original or updated library from a training perspective.

9. How the Ground Truth for the Training Set Was Established

The document implies that the reference library is built from well-characterized and known organisms. For the specific organisms added to the claimed list (Table 5), they represent further development/validation. The initial development of the database entries would have likely involved:

Definitive microbiological identification methods: Classic biochemical tests, microscopy, and, critically, molecular sequencing (e.g., 16S rRNA sequencing), which is mentioned as the ground truth method elsewhere (e.g., Acinetobacter baumannii complex resolution, Method Comparison).
The "Matching Algorithm" calculates matches by comparing a new spectrum against each single reference entry of a reference database. This suggests that each entry in the database itself originates from a definitively identified organism whose mass spectrum was collected and included.

Summary

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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 (800) 638 2041 or (301) 796-7100 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 (800) 638-2041 or (301) 796-7100 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 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 GmbHFahrenheitstrasse 428359 Bremen, Germany
Correspondent	Bruker Daltonik GmbHMarkus Kostrzewa, Vice President Clinical Mass Spectrometry R&DFahrenheitstrasse 428359 Bremen, GermanyPhone: +49 (0)421-2205 1258Fax: +49 (0)421-2205 1400Email: Markus.Kostrzewa@bruker.com
Device Identification
Trade or Proprietary Name:	MALDI Biotyper CA System
Common or Usual Name:	System, mass spectrometry, maldi tof, microorganism identification,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).

Table 1: Substantial Equivalency Table

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

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	ReproducibilityPanel	Site A:MBT-CA ID(DT+eDT+Ext)	Site B:MBT-CA ID(DT+eDT+Ext)	Site C:MBT-CA ID(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 negative aerobic, Gram positive aerobic / anaerobic bacteria and yeast were analysed. All sample preparation techniques (DT, eDT and Ext) were conducted. The rate of MBT-CA false identifications was 0% for this test. The overall identification rate of test samples at high confidence level (log(score) ≥2.0) was 100%.

General performance of the MBT Biotarget 96 US IVD and the US IVD MSP 48 Target Polished Steel was comparable.

Technical Study - Mass Accuracy / Target Edge Effects

In this study, approximately one thousand five hundred (1500) measurements were carried out. This study was designed for validation of the MBT Biotarget 96 US IVD in terms of "mass accuracy / target flatness" using BTS, MBT-CA System software and reference library and at least two (2) microflex LT/SH mass spectrometer systems. Validation was performed on five (5) MBT Biotarget 96 US IVD and five (5) US IVD MSP 48 Target Polished Steel. Targets were entirely prepared with dissolved BTS, dried and then overlaid with HCCA matrix.

Two parameters were evaluated during this study: Target Flatness (the mass accuracy was used as an indicator for target flatness) and edge effects (comparison of mass accuracy from edge positions vs inner target positions). For evaluation, one prominent reference protein (e.g. 6255.4 m/z) within the mass range of the BTS was analyzed concerning mass reproducibility. Additionally the log(scores) of each BTS spectrum were calculated as an indicator of identification performance.

Generally it can be stated that the technical performance between the MBT Biotarget 96 US IVD compared to the US IVD MSP 48 Target Polished Steel was comparable.

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Equivalence Study MBT Biotarget 96 US IVD (Smartbeam Laser)

The study design of this study was identical to the previous chapter [Equivalence Study MBT Biotarget 96 US IVD (Nitrogen Laser)]. In contrast to the 10,000 measurements in the previous chapter using two nitrogen laser systems, in this study an additional 5000 measurements were performed on one MBT smart CA System (Smartbeam Laser) to further prove the equivalence of both target types.

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
. ldentification of 50 FDA cleared organisms according to MBT-CA System workflow.

Generally it can be stated that the technical performance and equivalence between the MBT Biotarget 96 US IVD compared to the US IVD MSP 48 Target Polished Steel could be shown again in this study.

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Nocardia Study

This study was designed to verify and validate the measurement and identification of different strains and different species of the genus Nocardia by using the MBT-CA System as well as the MBT smart CA System in parallel. In addition the study was performed using the US IVD MSP 48 Target Polished Steel and the MBT Biotarget 96 US IVD in parallel. The results of this study can be evaluated as analytical study for identification performance of the MBT-CA System for the genus Nocardia and additionally as demonstration of equivalence between the "MBT-CA and MBT smart CA Systems" as well as equivalence of the "MBT Biotarget 96 US IVD and US IVD MSP 48 Target Polished Steel".

Analytical Studies:

Measurement of 30 strains covering 6 Nocardia species.
DT, eDT and Ext was used for sample preparation. ●
Each sample preparation technique was spotted 8 times on the targets. ●
Each spot was measured twice.
. MALDI targets "US IVD MSP 48 Target Polished Steel" and "MBT Biotarget 96 US IVD" were used.
Measurement on MBT smart CA System and MBT-CA System.
. Nocardia were measured after 24h, 48h and after 5 days of cultivation.

Altogether about 15,000 measurements were performed (30 strains * 3 sample preparations * 8 target spots * 2 measurements per spot * 2 target types * 2 instrument types * three cultivation times ). The identification performance of Nocardia using the standard identification MBT-CA workflow was excellent. The general log(score) distribution was similar and the the equivalence for all examined components (MBT-CA System, MBT smart CA System, MBT Biotarget 96 US IVD, and US IVD MSP 48 Target Polished Steel) was shown successfully.

Image /page/24/Figure/13 description: The image shows a graph titled "OVERALL PERFORMANCE 48h" with log(score) on the y-axis ranging from 0.00 to 3.00. The x-axis is divided into sections labeled "Nitrogen", "Smart", "Nitrogen", and "Smart", each associated with either "Biotarget" or "Steel Target". Within each section, there are data points for "DT", "eDT", and "Ext", with the data points appearing as red clusters.

As an example all 5760 log(scores) after 48h cultivation are shown in the following diagram:

Figure 1: 5760 spectra derived log(scores) after 48h cultivation overlaid. Each "log(score) lane" represents 480 single log(scores). The "focal points" are very similar.

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Conclusion:

Studies found that the MBT smart System and the MBT System, as well as the MBT Biotarget 96 US IVD and US IVD MSP 48 Target Polished Steel had comparable performance.

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Method Comparison:

To demonstrate performance of the MALDI Biotyper CA (MBT-CA) System, a method comparison study was performed at four (4) US clinical test sites and in-house laboratory. 2,091 fresh and stored organisms were tested on the MALDI Biotyper CA System in accordance with manufacturer's instructions for use. All organisms included in the study were sub-cultured for purity. Testing on the MBT-CA System was done from a freshly isolated colony.

All organisms included in the study and tested by US study sites were also sub-cultured onto an agar slant or appropriate media for isolation and shipped to the study interim reference laboratory. The interim reference laboratory stored all organisms included in the study and when requested, sent organisms to the sequencing reference laboratory for 16S rRNA or ITS sequencing and protein gene sequencing when requested. In-house laboratory and some testing sites processed their own isolates to the sequencing reference laboratory.

The following Gram-negative, Gram-positive and yeast species are additionally included in the reference library (please refer to K130831 and K142677 for previously claimed organisms).

Organisms
Abiotrophia defectiva
Acinetobacter_baumannii_nosocomialis group
Acinetobacter calcoaceticus
Acinetobacter pittii
Actinomyces europaeus
Actinomyces funkei
Actinomyces graevenitzii
Actinomyces hyovaginalis
Actinomyces radingae
Actinomyces turicensis
Actinomyces urogenitalis
Actinotignum_schaalii group
Aerococcus sanguinicola
Aggregatibacter actinomycetemcomitans
Aggregatibacter aphrophilus
Aggregatibacter segnis
Alloiococcus otitis
Alloscardovia omnicolens
Anaerococcus murdochii
Arthrobacter cumminsii
Bacteroides caccae
Bacteroides nordii
Bacteroides pyogenes
Bacteroides salversiae
Organisms
Bacteroides stercoris group
Bifidobacterium breve
Candida intermedia
Candida zeylanoides
Clostridium beijerinckii
Clostridium bifermentans
Clostridium butyricum
Clostridium_clostridioforme group
Clostridium innocuum
Clostridium paraputrificum
Clostridium ramosum
Clostridium septicum
Clostridium sordellii
Clostridium sporogenes/ Clostridium botulinum (group I)
Clostridium tertium
Corynebacterium accolens
Corynebacterium_afermentans group
Corynebacterium coyleae
Corynebacterium freneyi
Corynebacterium glutamicum
Corynebacterium_mucifaciens_ureicelerivorans group
Corynebacterium pseudotuberculosis
Corynebacterium resistens
Cyberlindnera jadinii
Dermabacter hominis
Enterococcus durans
Enterococcus mundtii
Enterococcus raffinosus
Escherichia hermannii
Escherichia vulneris
Ewingella americana
Facklamia hominis
Fluoribacter bozemanae
Gemella morbillorum
Helcococcus kunzii
Kingella denitrificans
Klebsiella variicola
Lactobacillus gasseri
Lactobacillus jensenii
Lactobacillus rhamnosus
Leclercia adecarboxylata
Legionella longbeachae
Legionella pneumophila
Organisms
Leuconostoc pseudomesenteroides
Listeria monocytogenes
Malassezia furfur
Malassezia pachydermatis
Mannheimia_haemolytica group
Micrococcus lylae
Mobiluncus curtisii
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
Parabacteroides goldsteinii
Parabacteroides_johnsonii_merdae group
Parvimonas micra
Pediococcus acidilactici
Pluralibacter gergoviae
Porphyromonas somerae
Ralstonia pickettii
Rhodotorula mucilaginosa
Serratia fonticola
Serratia odorifera
Sphingobacterium multivorum
Sphingobacterium spiritivorum
Sphingomonas paucimobilis
Staphylococcus delphini
Staphylococcus intermedius
Staphylococcus lentus
Staphylococcus sciuri
Staphylococcus xylosus
Streptococcus canis
Streptococcus equi
Streptococcus parasanguinis

Table 5: Claimed Organisms

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Table 6 to Table 8 below show the overall isolate performance.

Table 6: Overall Isolate Performance

Overall Performance - CLAIM 3
MBT-CA RESULT	REFERENCE ALGORITHM
	high resolutionspecies	low resolutionspecies / genus	Negative	Total
Organism ID ≥ 2.0(High Confidence)	1904	130	21	2055
Organism ID (≥1.7; <2.0)(Low Confidence)	23	5	4	32
- INCORRECT MBT-CA ID (≥1.7)- NO ID (<1.7)	3	1	n/a	4
Total	1930	136	25	2091

Positive	Negative
highresolution	high & lowresolution
98.45%	99.81%	n/a

Table 7: Overall Bacteria Performance

Performance BACTERIA
MBT-CA RESULT	REFERENCE ALGORITHM
	high resolutionspecies	low resolutionspecies / genus	Negative	Total
Organism ID ≥ 2.0(High Confidence)	1827	130	21	1978
Organism ID (≥1.7; <2.0)(Low Confidence)	21	5	4	30
- INCORRECT MBT-CA ID (≥1.7)- NO ID (<1.7)	2	1	n/a	3
Total	1850	136	25	2011

Positive		Negative
highresolution	high & lowresolution	n/a
98.54%	99.85%	n/a

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Page 22 of 22 June 21, 2017

Table 8: Overall Yeast Performance

Performance YEAST
MBT-CA RESULT	REFERENCE ALGORITHM
	high resolutionspecies	low resolutionspecies / genus	Negative	Total
Organism ID ≥ 2.0(High Confidence)	77	0	0	77
Organism ID (≥1.7; <2.0)(Low Confidence)	2	0	0	2
- INCORRECT MBT-CA ID (≥1.7)- NO ID (<1.7)	1	0	n/a	1
Total	80	0	0	80

Positive	Negative
highresolution	high & lowresolution
96.25%	98.75%	n/a

Statement of Safety and Efficacy

The data presented clearly demonstrate the safety and efficacy of the MBT-CA System as compared to the reference algorithm, when the instructions for use are followed.

Regulation Number and Section

§ 866.3361 Mass spectrometer system for clinical use for the identification of microorganisms.

(a)
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.(b)
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 as part of your 21 CFR 820.30 design controls, 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.

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 massspectrometer systems using matrix-assistedlaser desorption/ionization - time of flight(MALDI-TOF) for the identification ofmicroorganisms cultured from humanspecimens.The MALDI Biotyper CA System is aqualitative in vitro diagnostic devicesindicated for use in conjunction with otherclinical and laboratory findings to aid in thediagnosis of bacterial and yeast infections.	The MALDI Biotyper CA System is a massspectrometer system using matrix-assistedlaser desorption/ionization - time of flight(MALDI-TOF) for the identification ofmicroorganisms cultured from humanspecimens.The MALDI Biotyper CA System is aqualitative in vitro diagnostic deviceindicated for use in conjunction with otherclinical and laboratory findings to aid in thediagnosis of bacterial and yeast infections.
Sample type	Isolated colony from any patient samplesource.Acceptable media:• Columbia blood agar with 5% sheepblood (Gram-negative bacteria)• Trypticase soy agar with 5% sheep blood(Gram-negative bacteria, Gram-positivebacteria, yeasts)• Chocolate agar (Gram-negative bacteria,Gram-positive bacteria)• MacConkey Agar (Gram-negativebacteria)• Columbia CNA agar with 5% sheep blood(Gram-positive bacteria)• Brucella Agar with 5% horse blood (Gram-negative anaerobic bacteria, Gram-positive anaerobic bacteria)• CDC anaerobe Agar with 5% sheep blood(Gram-negative anaerobic bacteria,Gram-positive anaerobic bacteria)• CDC anaerobe 5% sheep blood Agar withphenylethyl alcohol (Gram-negativeanaerobic bacteria, Gram-positiveanaerobic bacteria)• CDC anaerobe laked sheep blood Agarwith kanamycin and vancomycin (Gram-negative anaerobic bacteria)• Bacteroides bile esculin Agar withamikacin ( Bacteroides species)	Isolated colony from any patient samplesource.Acceptable media:• Columbia blood agar with 5% sheepblood (Gram-negative bacteria)• Trypticase soy agar with 5% sheep blood(Gram-negative bacteria, Gram-positivebacteria, yeasts)• Chocolate agar (Gram-negative bacteria,Gram-positive bacteria)• MacConkey Agar (Gram-negativebacteria)• Columbia CNA agar with 5% sheep blood(Gram-positive bacteria)• Brucella Agar with 5% horse blood (Gram-negative anaerobic bacteria, Gram-positive anaerobic bacteria)• CDC anaerobe Agar with 5% sheep blood(Gram-negative anaerobic bacteria,Gram-positive anaerobic bacteria)• CDC anaerobe 5% sheep blood Agar withphenylethyl alcohol (Gram-negativeanaerobic bacteria, Gram-positiveanaerobic bacteria)• CDC anaerobe laked sheep blood Agarwith kanamycin and vancomycin (Gram-negative anaerobic bacteria)• Bacteroides bile esculin Agar withamikacin ( Bacteroides species)
Similarities
	NEW DEVICEMBT-CA System (K163536)	PRIMARY PREDICATE DEVICEMBT-CA System (K142677)
Characteristic	blood ( Clostridium difficile )• Sabouraud-Dextrose Agar (Yeasts)• Brain Heart Infusion Agar (Yeasts)• Campylobacter Agar with 5 Antimicrobicsand 10% Sheep Blood ( Campylobacterspecies)• Bordet Gengou Agar with 15% sheepblood ( Bordetella species)• plus three (3) additionally validatedmedia (see differences).	blood ( Clostridium difficile )• Sabouraud-Dextrose Agar (Yeasts)• Brain Heart Infusion Agar (Yeasts)• Campylobacter Agar with 5 Antimicrobicsand 10% Sheep Blood ( Campylobacterspecies)• Bordet Gengou Agar with 15% sheepblood ( Bordetella species)
Type of Test	Automated Mass Spectrometry System	Automated Mass Spectrometry System
Matrix	α-Cyano-4-hydroxycinnamic acid	α-Cyano-4-hydroxycinnamic acid
Method ofTesting	Bacteria & Yeast: Direct testingIf after initial analysis the log(score) isreported at <2.00, organisms may beprocessed using the Extraction (Ext)procedure or extended Direct Transfer (eDT,70% aqueous formic acid) procedure. If eDTprocedure still yields log(score) <2.00,organisms may be processed via Extprocedure.	Bacteria & Yeast: Direct testingIf after initial analysis the log(score) isreported at <2.00, organisms may beprocessed using the Extraction (Ext)procedure or extended Direct Transfer (eDT,70% aqueous formic acid) procedure. If eDTmethod still yields log(score) <2.00,organisms may be processed via Extprocedure.
Result Reporting	Organism identification is reported withhigh confidence if the log(score) is ≥2.00.An organism identification is reported withlow confidence if the log(score) is between1.70 and <2.00.	Organism identification is reported withhigh confidence if the log(score) is ≥2.00.An organism identification is reported withlow confidence if the log(score) is between1.70 and <2.00.
MatchingAlgorithm	Calculates matches by comparing a newspectrum against each single referenceentry of a reference database.	Calculates matches by comparing a newspectrum against each single referenceentry of a reference database.
Recorded massrange	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 forClinical Applications (MBT-CA) - update	MALDI Biotyper Reference Library forClinical Applications (MBT-CA)