Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The description details a standard immunoassay using a competitive binding principle and a calibration curve. There is no mention of AI or ML in the device description, intended use, or performance studies.

Therapeutic

No.
This device is an in vitro diagnostic (IVD) assay used to quantitate vitamin B12 for diagnosis and treatment of malabsorption, it does not provide therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that "Measurements obtained by this device are used in the diagnosis and treatment of gastrointestinal malabsorption." This directly indicates its role as a diagnostic device.

SaMD (Software as a Medical Device)

No

The device description explicitly details physical components like reagents, microparticles, and calibrators, which are hardware components used in the immunoassay process. The device is an in vitro diagnostic (IVD) assay that runs on specific immunoassay analyzers, which are also hardware.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use/Indications for Use: The document explicitly states "Binding assay for the in vitro quantitation of vitamin B12 in human serum and plasma." and "Measurements obtained by this device are used in the diagnosis and treatment of gastrointestinal malabsorption." This clearly indicates the device is intended for use on biological samples (serum and plasma) outside of the body ("in vitro") for diagnostic purposes.
Device Description: The description details the components and principles of an immunoassay designed to measure a substance (vitamin B12) in a biological sample.
Performance Studies: The document includes detailed performance studies (Precision, Limit of Blank, Limit of Detection, Method Comparison, etc.) which are standard for IVD devices to demonstrate their analytical performance.
Predicate Device: The mention of a "Predicate Device" (K060755; Elecsys Vitamin B12 Assay) is a strong indicator that this device is being submitted for regulatory review as an IVD, often through a 510(k) pathway where comparison to a legally marketed predicate IVD is required.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Binding assay for the in vitro quantitation of vitamin B12 in human serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of gastrointestinal malabsorption. The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassay analyzers.

Vitamin B12 II CalSet is used for calibrating the quantitative Elecsys Vitamin B12 II assay on the Elecsys and cobas e immunoassay analyzers.

Product codes (comma separated list FDA assigned to the subject device)

CDD, JIT

Device Description

The Elecsys Vitamin B12 II assay employs a competitive test principle using intrinsic factor specific for vitamin B12. Vitamin B12 in the sample competes with the added vitamin B12 labeled with biotin for the binding sites on the ruthenium-labeled intrinsic factor complex.

Results are determined via a calibration curve which is instrument specifically generated by 2-point calibration and a master curve provided via reagent barcode.

The reagent working solutions include:

the rackpack (kit placed on instrument) .
Streptavidin coated microparticles, .
Reagent 1 (ruthenium labeled intrinsic factor) and .
Reagent 2 (vitamin B12 labeled biotin). .
Pretreatment 1 (Dithiothreitol) .
Pretreatment 2 (sodium hydroxide, sodium cyanide) .

The Vitamin B12 II CalSet is a lyophilized human serum matrix with added vitamin B12 in two concentration ranges.

The CalSet includes:

Cal 1 (approximately 250 pg/mL vitamin B12 in a Human serum matrix) .
Cal 2 (approximately 1500 pg/mL vitamin B12 in a Human serum matrix) .

Mentions image processing

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Mentions AI, DNN, or ML

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Input Imaging Modality

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Anatomical Site

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Indicated Patient Age Range

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Intended User / Care Setting

Prescription Use (Part 21 CFR 801 Subpart D)

Description of the training set, sample size, data source, and annotation protocol

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Description of the test set, sample size, data source, and annotation protocol

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Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

4.1. Precision

Study Type: Precision
Sample Size: Not explicitly stated, but involved serum samples, 2 replicates per run, 2 runs per day for 21 days for within-run and intermediate precision.
Key Results: All results met the pre-defined acceptance criteria for repeatability and intermediate precision.
- Specifications:
  - Repeatability: ≤ 200 pg/mL: SD ≤ 14 pg/mL; > 200 pg/mL: CV ≤ 7 %
  - Intermediate precision: ≤ 200 pg/mL: SD ≤ 24 pg/mL; > 200 pg/mL: CV ≤ 12 %

4.2. Limit of Blank

Study Type: Analytical sensitivity (LoB)
Sample Size: n = 60 LoB measurements (5 replicates, 2 runs per day on 2 instruments over 3 days).
Key Results: LoB ≤ 50 pg/mL (Acceptance criterion)

4.3. Limit of Detection

Study Type: Analytical sensitivity (LoD)
Sample Size: n = 60 LoD measurements (5 samples, 2 runs per day on 2 instruments over 3 days).
Key Results: LoD ≤ 100 pg/mL (Acceptance criterion)

4.4. Limit of Quantitation

Study Type: Limit of Quantitation (LoQ)
Sample Size: Minimum set of seven human serum samples, three reagent lots. Each sample analyzed in replicates of 5, one run per day over 5 days.
Key Results: LoQ (Imprecision) ≤ 20 % at 150 pg/mL (Acceptance criterion)

4.5. Dilution

Study Type: Dilution Study
Sample Size: Five serum samples.
Key Results: Recovery was investigated, and undiluted concentrations were calculated by multiplying diluted results with the appropriate dilution factor.

4.6. Linearity

Study Type: Linearity
Sample Size: Not explicitly stated, but involved serum samples.
Key Results: All results met the predefined acceptance criteria for linearity. The results support a claimed measuring range.
- Acceptance criteria:
  - Significant level for deviation to higher order polynomial: 5%
  - Limits for deviation to higher order polynomial regression:
    - ≤ 200 pg/mL: ± 20 pg/mL
    - 200 pg/mL: ± 10 %

4.7. Analytical Specificity

Study Type: Analytical Specificity (Cross-reactivity)
Sample Size: Two human serum samples, four levels of cross-reactant. Tested in duplicate.
Key Results: Will be reported in the method sheet.

4.8. Endogenous Interferences

Study Type: Endogenous Interferences
Sample Size: Three human serum samples (low, mid, high concentrations of Vitamin B12) for each interfering substance (Biotin, Lipemia, Hemoglobin, Bilirubin, Rheumatoid Factor, IgG/IgM/IgA).
Key Results: Predefined acceptance criterion was met. Claims included in the method sheet were set to the concentration without observed interference.
- Acceptance criterion:
  - 200 pg/mL: ± 10% of unspiked reference value

4.9. Exogenous Interferences- Drugs

Study Type: Exogenous Interferences (Drugs)
Sample Size: Two human serum samples spiked with 16 pharmaceutical compounds.
Key Results: Acceptance criterion was ± 10% of the reference value (unspiked sample).

4.10. Exogenous Interferences- Anticoagulants

Study Type: Exogenous Interferences (Anticoagulants)
Sample Size: Minimum of 90 serum/plasma pairs per sample material.
Key Results: Potential effects assessed by regression analysis.
- Acceptance criterion:
  - Slope (Passing/Bablok): 0.9 – 1.1
  - Intercept (Passing/Bablok): 0.95
  - Intercept (Passing/Bablok): 200 pg/mL: ± 14 %

4.14. Calibrator studies

Study Type: Calibrator studies (Value assignment, Reconstitution, Stability)
- 4.14.1. Value assignment: Target values chosen for best fit with Master Calibration Curve. PreciControl Varia acceptance criteria must be met.
- 4.14.2. Reconstitution: Signal recovery after 30 minutes reconstitution compared to 15 minutes.
- 4.14.3. Stability: Three studies performed (Stability at -20ºC after reconstitution, Stability at 2 to 8°C after reconstitution, Onboard stability at 20-25℃ after reconstitution). Real-time stability data of predicate device is applicable.
Sample Size: Not explicitly stated for all, but for value assignment, calibrators run in duplicate on at least three (3) cobas e 411 analyzers and at least three (3) cobas e 601/MODULAR ANALYTICS E170 analyzers.
Key Results: Value assignment passed. Reconstitution acceptance criterion was 90 to 110 % signal recovery of the reference material value. Stability studies acceptance criterion was 90 to 110 % signal recovery of the reference material value.

5. CLINICAL PERFORMANCE EVALUATION

Study Type: Clinical Performance Evaluation (Reference Range Study)
Sample Size: Samples collected at four sites in the US for reference range establishment. Not explicitly given for total. 120 apparently healthy male & female subjects were tested in one specific reference range study comparison.
Key Results: Reference ranges for apparently healthy males and females were determined using median and 2.5th – 97.5th percentiles.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

LOD (Limit of Detection): ≤ 100 pg/mL
LoB (Limit of Blank): ≤ 50 pg/mL
LoQ (Limit of Quantitation): ≤ 20 % at 150 pg/mL (Imprecision)
Precision:

Repeatability: ≤ 200 pg/mL: SD ≤ 14 pg/mL; > 200 pg/mL: CV ≤ 7 %
Intermediate precision: ≤ 200 pg/mL: SD ≤ 24 pg/mL; > 200 pg/mL: CV ≤ 12 %
Linearity:
≤ 200 pg/mL: ± 20 pg/mL
200 pg/mL: ± 10 %
Accuracy (Method Comparison):
Slope (Passing/Bablok): 1.00 ± 0.05
Correlation (linear regression) Pearson's r: > 0.95
Intercept (Passing/Bablok): 200 pg/mL: ± 10% of unspiked reference value
Sample Stability Recovery:
≤ 200 pg/mL: ± 28 pg/mL
200 pg/mL: ± 14 %
Calibrator Studies Signal Recovery: 90 to 110 % of the reference material value.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Elecsys Vitamin B12 Assay (K060755)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

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Regulation Number and Section

§ 862.1810 Vitamin B

12 test system.(a)
Identification. A vitamin B12 test system is a device intended to measure vitamin B12 in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of anemias of gastrointestinal malabsorption.(b)
Classification. Class II.

Summary

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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 circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, stacked on top of each other.

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

ROCHE DIAGNOSTICS KELLI TÜRNER REGULATORY AFFAIRS PRINCIPAL 9115 HAGUE ROAD INDIANAPOLIS IN 46250

September 24, 2015

Re: K151786

Trade/Device Name: Elecsys Vitamin B12 II Assay, Elecsys Vitamin B12 II CalSet Regulation Number: 21 CFR 862.1810 Regulation Name: Vitamin B12 test system Regulatory Class: II Product Code: CDD, JIT Dated: June 29, 2015 Received: July 1, 2015

Dear Ms. Turner:

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 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.

1

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/Resourcesfor You/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.

Katherine Serrano -S

For : Courtney H. Lias Director Division of Chemistry and Toxicology 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 : K151786

Device Name Elecsys Vitamin B12 II assay

Indications for Use (Describe)

Binding assay for the in vitro quantitation of vitamin B12 in human serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of gastrointestinal malabsorption. The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassay analyzers.

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.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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Indications for Use

510(k) Number:K151786

Device Name Elecsys Vitamin B12 II CalSet

Indications for Use (Describe)

Vitamin B12 II CalSet is used for calibrating the quantitative Elecsys Vitamin B12 II assay on the Elecsys and cobas e immunoassay analyzers.

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

X Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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Elecsys Vitamin B12 II Test System 510(k) Summary:K151786

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

Submitter Name	Roche Diagnostics
Address	9115 Hague Road
Indianapolis, IN 46250
Contact	Kelli Turner
Phone: (317) 521-4515
FAX: (317) 521-2324
Email: kelli.turner@roche.com
Date Prepared	06/29/2015
Proprietary Name	1. Elecsys Vitamin B12 II assay,

Elecsys Vitamin B12 II CalSet |
| Common Name | 1. Vitamin B12 II assay
Vitamin B12 II CalSet |
| Classification Name | 1. Radioassay, Vitamin B12
Secondary, calibrator |
| Product Codes | 1. CDD, 862.1810
JIT, 862.1150 |
| Predicate Devices | Elecsys Vitamin B12 Assay (K060755) |
| Establishment Registration | Roche Diagnostics GmbH in Mannheim, Germany, is 9610126
Roche Diagnostics GmbH in Penzberg, Germany, is 9610529
Roche Diagnostics in the United States is 1823260 |

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1. DEVICE DESCRIPTION

The Elecsys Vitamin B12 II assay employs a competitive test principle using intrinsic factor specific for vitamin B12. Vitamin B12 in the sample competes with the added vitamin B12 labeled with biotin for the binding sites on the ruthenium-labeled intrinsic factor complex.

Results are determined via a calibration curve which is instrument specifically generated by 2-point calibration and a master curve provided via reagent barcode.

1.1. Reagents

The reagent working solutions include:

the rackpack (kit placed on instrument) .
Streptavidin coated microparticles, .
Reagent 1 (ruthenium labeled intrinsic factor) and .
Reagent 2 (vitamin B12 labeled biotin). .
Pretreatment 1 (Dithiothreitol) .
Pretreatment 2 (sodium hydroxide, sodium cyanide) .

Calibrator 1.2.

The Vitamin B12 II CalSet is a lyophilized human serum matrix with added vitamin B12 in two concentration ranges.

The CalSet includes:

Cal 1 (approximately 250 pg/mL vitamin B12 in a Human serum matrix) .
Cal 2 (approximately 1500 pg/mL vitamin B12 in a Human serum matrix) .

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2. INDICATIONS FOR USE

Elecsys B12 II assay

Binding assay for the in vitro quantitative determination of vitamin B12 in human serum and plasma. Measurements obtained by this device are used in the diagnosis and treatment of anemias of gastrointestinal malabsorption.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on Elecsys and cobas e immunoassay analyzers.

Elecsys B12 II CalSet

Vitamin B12 II CalSet is used for calibrating the quantitative Elecsys Vitamin B12 II assay on the Elecsys and cobas e immunoassay analyzers.

TECHNOLOGICAL CHARACTERISTICS 3.

Table 1: Assay Comparison: General Assay Features

| Feature | Predicate Device: Elecsys Vitamin B12
(K060755) | Candidate Device: Elecsys Vitamin B12 II Assay |
|-----------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use/
Indications for
Use | Binding assay for the in vitro quantitative
determination of vitamin B12 in human serum
and plasma.
The electrochemiluminescence immunoassay
"ECLIA" is intended for use on Elecsys and
cobas e immunoassay analyzers.
A Vitamin B12 test system is a device intended
to measure Vitamin B12 in serum, plasma.
Measurements obtained by this device are
used in the diagnosis and treatment of anemias
of gastrointestinal malabsorption. | Binding assay for the in vitro quantitative
determination of vitamin B12 in human serum
and plasma. Measurements obtained by this
device are used in the diagnosis and treatment
of anemias of gastrointestinal malabsorption.
The electrochemiluminescence immunoassay
"ECLIA" is intended for use on Elecsys and
cobas e immunoassay analyzers. |
| Assay Protocol | The Elecsys Vitamin B12 assay employs a
competitive test principle using
intrinsic factor specific for vitamin B12. Vitamin
B12 in the sample competes with the added
vitamin B12 labeled with biotin for the binding
sites on the ruthenium- labeled intrinsic factor
complex. | Same. |
| Detection
Protocol | Electrochemiluminescent Assay | Same. |
| Applications | 27-minute application | Same. |
| Instrument
Platform | Elecsys and cobas e immunoassay analyzers. | Same. (submission for cobas e 411) |
| Sample Volume | 15 µL | Same. |
| Sample
Type | Human serum and plasma. | Same. |
| Reagents | Competition principle. Total duration of assay:
27 minutes | Same. |
| Calibrator | Elecsys Vitamin B12 CalSet II | Elecsys Vitamin B12 II CalSet |

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| Feature | Predicate Device: Elecsys Vitamin B12
(K060755) | Candidate Device: Elecsys Vitamin B12 II
Assay |
|-----------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Calibration
Interval | Calibration must be performed once per
reagent lot using fresh reagent (i.e. not more
than 24 hours since the reagent kit was
registered on the analyzer). Renewed
calibration is recommended as follows:
• After 1 month (28 days) when using the
same reagent lot.
• After 7 days (when using the same
reagent kit on the analyzer).
• As required: e.g. quality control findings
outside the defined limits | Same. |
| Controls | Elecsys PreciControl Varia | Same |
| Traceability /
Standardization | Standardized against the Elecsys Vitamin B12
assay (K973702) | Standardized against the Elecsys Vitamin B12
assay (K060755).
Accuracy to WHO Standard 03/178 |
| Reagent
Stability | Unopened: 2-8°C - Up to the stated expiration
date.
After Opening at 2-8°C - 12 weeks
On the Analyzers – 5 weeks | Unopened: 2-8°C - Up to the stated expiration
date.
After Opening at 2-8°C - 12 weeks
On the Analyzers - 5 weeks onboard or 60
days when stored alternatively in the
refrigerator and on the analyzer, with the total
time onboard on the analyzer not exceeding 10
x 8 hours |
| Measuring
Range | 30.0 - 2000 pg/mL | 150- 2000 pg/mL |

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| Feature (K060755) | | | Candidate Device: Elecsys Vitamin B12 II Assay | | | | |
-----------------------------------------------|------------------------------------|----------------------------------------------|-----------------------------------------------------------------------------------------------------------------------|----------------|------------------------------------|----------------------------------------------|------|
| cobas e 411:
| | cobas e 411:
| | | | |
| Mean (pg/mL) | SD | CV | Sample | Mean (pg/mL) | SD | CV |
| 192 | 11.3 | 5.9% | HS 1 | 176 | 8.86 | 5.0% |
| 358 | 20.1 | 5.6% | HS 2 | 405 | 13 | 3.2% |
| 864 | 29.1 | 3.3% | HS 3 | 960 | 19.7 | 2.1% |
| 1625 | 62.1 | 3.8% | HS 4 | 1230 | 27.4 | 2.2% |
| 221 | 10.2 | 4.6% | HS 5 | 1940 | 40.9 | 2.1% |
| 467 | 18.5 | 4.0% | PCV0 | 229 | 8.96 | 3.9% |
| 958 | 31.1 | 3.3% | PCV1 | 447 | 12.2 | 2.7% |
| | | | PCV2 | 934 | 20.2 | 2.2% |
| | | Total (will be labeled Intermediate precision) | | | | |
| Mean (pg/mL) | SD | CV | Sample | Mean (pg/mL) | SD | CV |
| 192 | 19.8 | 10.3% | HS 1 | 176 | 12.7 | 7.2% |
| 358 | 27.7 | 7.7% | HS 2 | 405 | 17.5 | 4.3% |
| 864 | 40.1 | 4.5% | HS 3 | 960 | 31.0 | 3.2% |
| 1625 | 65.1 | 4.0% | HS 4 | 1230 | 46.4 | 3.8% |
| 221 | 10.2 | 4.6% | HS 5 | 1940 | 72.6 | 3.7% |
| 467 | 18.5 | 4.0% | PCV0 | 229 | 12.4 | 5.4% |
| 958 | 31.1 | 3.3% | PCV1 | 447 | 18.6 | 4.2% |
| | | | PCV2 | 934 | 38.4 | 4.1% |
limit = 30.0 pg/mL | | | Limit of Blank (LoB): = 50 pg/mL Limit
| | | |
| concentration
| Cross reactant | concentration
| 200 | 0.024 | | Cobinamide | 210 | 0.003 | |
| 30 to 2000 pg/mL | | | 150 to 2000 ng/mL | | | | |

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| Feature | Predicate Device: Elecsys Vitamin B12
(K060755) | Candidate Device: Elecsys Vitamin B12 II
Assay |
|-------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Limitations | The assay is unaffected by:
• Bilirubin 200 pg/mL | CV ≤ 7 % | CV ≤ 12 % |

All results met the pre-defined acceptance criteria for repeatability and intermediate precision.

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4.2. Limit of Blank

For the analytical sensitivity studies, 2 cobas e 411 analyzers and 3 lots of reagents were used. The Limit of Blank (LoB) was determined using a buffered human serum albumin matrix similar to human serum (no analyte added). A total of n = 60 LoB measurements were made (5 replicates, 2 runs per day on 2 instruments over 3 days). The LoB was calculated according to CLSI EP17-A2 (non-parametric approach). Acceptance criterion: LoB ≤ 50 pg/mL

Limit of Detection 4.3.

For the analytical sensitivity studies, 2 cobas e 411 analyzer and 3 lots of reagents were used. The Limit of Detection (LoD) was determined using 5 low-level human serum samples (native and diluted). A total of n = 60 LoD measurements were made (5 samples, 2 runs per day on 2 instruments over 3 days). The LoD was calculated according to CLSI EP17-A2 (chapter 5.3.3.2). Acceptance criterion: LoD ≤ 100 pg/mL

Limit of Quantitation 4.4.

The Limit of Quantitation (LoQ) was determined using a minimum set of seven human serum samples, three reagent lots on one cobas e 411 analyzer. The Limit of Quantitation (LoQ) was determined in accordance with CLSI Guideline EP17-A2. Each sample was analyzed in replicates of 5, one run per day over 5 days. Acceptance criterion: LoQ (Imprecision) ≤ 20 % at 150 pg/mL.

4.5. Dilution

To demonstrate the Vitamin B12 II assay dilution study, four different dilutions with dilution factors between 1:1.5 and 1:3 were prepared.

Five serum samples were spiked with vitamin B12 to concentrations exceeding the measuring range. The samples were diluted with Elecsys Diluent Universal and recovery was investigated on the cobas e 411. The vitamin B12 concentrations of the undiluted samples were calculated by multiplying the result of the diluted samples using the appropriate dilution factor.

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4.6. Linearity

The linearity results were obtained with serum samples on the cobas e 411 Immunoassay analyzer. Linearity was determined according to CLSI Guideline EP6-A. All results met the predefined acceptance criteria for linearity. The linearity results support a claimed measuring range.

Acceptance criteria:

Significant level for deviation to higher order polynomial: 5% .
Limits for deviation to higher order polynomial regression .
- ≤ 200 pg/mL: ± 20 pg/mL .
- 200 pg/mL: ± 10 % .

4.7. Analytical Specificity

The specificity was determined using two human serum samples (single donors, native) spiked with potential cross-reactant compounds. The analyte concentration of the samples was at 129 and 550 pg/mL Vitamin B12. Four levels of the cross-reactant were prepared and measured in duplicate. The spiked and non-spiked samples were tested in duplicates on the cobas e 411 Immunoassay Analyzer. Cross-reactivity results will be reported in the method sheet.

Endogenous Interferences 4.8.

The effect on quantitation of analyte in the presence of endogenous interfering substances was determined on the cobas e 411 Immunoassay analyzer using human serum samples (single donors, native as well as spiked). For each interfering substance (Biotin, Lipemia, Hemoglobin, Bilirubin, Rheumatoid Factor, IgG/IgM/IgA) three human serum samples containing low, mid and high concentrations of Vitamin B12 were tested. The recovery for each sample was calculated by comparison to the reference sample. Predefined acceptance criterion was met. The claims included in the method sheet were set set to the concentration without observed interference.

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Acceptance criterion:

. 200 pg/mL: ± 10% of unspiked reference value .

Exogenous Interferences- Drugs 4.9.

The effect on quantitation of analyte in the presence of drugs was determined by comparing values obtained from samples spiked with 16 pharmaceutical compounds spiked into two human serum samples (single donors, native) and tested on the cobas e 411 Immunoassay Analyzer. The analyte concentration of the samples were approximately 200 and 1200 pg/mL. The drug concentrations tested are in accordance with the recommendation (if available) given in the CLSI guideline EP7-A2. When concentrations are not given in the guideline, concentrations of at least 3-times of the maximum recommended daily dose were tested. The two serum samples were divided into aliquots and spiked with the potential interferents. The reference sample without interferent was spiked with the respective amount of solvent only.

The concentration of the spiked aliquots was determined in 8-fold determination and compared to the Vitamin B12 concentration determined for the reference aliquot (also in 8-fold determination). Acceptance criterion: ± 10% of the reference value (unspiked sample).

4.10. Exogenous Interferences- Anticoagulants

The effect on quantitation of analyte in the presence of anticoagulants with the Elecsys Vitamin B12 II Immunoassay was determined by comparing values obtained from samples (single donors, native as well as spiked) drawn into Serum, Li-Heparin, Na-Heparin, K2-EDTA-, K3-EDTA-plasma primary tubes and Li-Heparin Plasma Gel Separation Tubes. A minimum of 90 serum/plasma pairs per sample material were tested in singleton with one reagent lot on one cobas e 411 Immunoassay Analyzer. Potential effects are assessed by regression analysis.

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Acceptance criterion:

. Slope (Passing/Bablok): 0.9 – 1.1
Intercept (Passing/Bablok): 0.95
Intercept (Passing/Bablok): 200 pg/mL: ± 14 % .

4.13.1. Study 1. Sample stability at 2-8°C

Five samples for each sample type (Serum, K2-EDTA-, K3-EDTA-, Li-Heparin- and Na-Heparin plasma) were aliquoted and measured after storage at 2-8℃ for 49 hours. Measurements were performed with three-fold determination and recovery was calculated as percent of the reference value.

4.13.2. Study 2. Sample stability at Room Temperature (15-25°C)

Five samples for each sample type (Serum. K2-EDTA-, K3-EDTA-, Li-Heparin- and Na-Heparin plasma) were aliquoted and measured after storage at 15-25°C for 3 hours. The aliguot for the reference value was stored at -80°C and measured at the same time point.

Measurements were performed with three-fold determination and recovery was calculated as percent of the reference value.

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4.13.3. Study 3. Sample stability -15 to -25°C

Five samples for each sample type (Serum, K2-EDTA-, K3-EDTA-, Li-Heparin- and Na-Heparin plasma) were aliquoted and measured after storage at -15 to -25℃ for 57 days. The aliquot for the reference value was stored at -80°C and measured at the same time point. Measurements were performed with three-fold determination and recovery was calculated as percent of the reference value.

4.14. Calibrator studies

The Elecsys Vitamin B12 II CalSet was evaluated for value assignment, reconstitution and stability.

4.14.1. Value assignment

Value assignment testing was conducted and passed pre-defined acceptance criteria. The target values for the two levels of the Vitamin B12 II CalSet kit are chosen to obtain the best fit with the Master Calibration Curve, together with the Rodbard curve parameters encoded in the reagent barcode. For each Elecsys Vitamin B12 II CalSet lot manufactured, the calibrators are run in duplicate on at least three (3) cobas e 411 analyzers and at least three (3) cobas e 601/MODULAR ANALYTICS E170 analyzers with all Vitamin B12 II reagent lots available. The assigned value of each calibrator is defined as the mean value obtained over at least six (6) runs on at least three (3) analyzers) of the respective calibrator.

Measurement values for PreciControl Varia (Levels 0, 1 & 2), a multi-analyte control recommended for use to monitor accuracy and precision of specified analytes, are read from the calibration curves generated. The pre-defined acceptance criteria for PreciControl Varia have to be met to release the Assigned Values for Vitamin B12 II CalSet.

4.14.2. Reconstitution

Reconstitution time for the lyophilized Vitamin B12 II CalSet was tested. Two sets of Vitamin B12 II CalSet were reconstituted, one for 15 minutes and the other for 30 minutes. Signal recovery after 30 minutes reconstitution was compared to the signal value after 15 minutes.

Vitamin B12 II CalSet was evaluated in duplicate on the cobas e 411 analyzer as a reference.

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The acceptance criterion was 90 to 110 % signal recovery of the reference material value.

4.14.3. Stability

Three studies were performed in order to verify the stability claims for the Vitamin B12 II CalSet. Stability studies after reconstitution and an accelerated stability study were completed on the cobas e 411.

4.14.3.1. Study 1- Stability at -20ºC (after reconstitution)

The on-test and reference materials were tested in duplicate on the cobas e 411. The on-test material was reconstituted and stored in closed vials for 85 days at -20°C. The on-test signal recovery was calculated as percent of the reference value.

The acceptance criterion was 90 to 110 % signal recovery of the reference material value.

4.14.3.2. Study 2- Stability at 2 to 8°C after reconstitution

The on-test and reference materials were tested in duplicate on the cobas e 411. The on-test material was reconstituted and stored in closed vials for 73 hours at 2-8°C. The on-test signal recovery was calculated as percent of the reference value.

The acceptance criterion was 90 to 110 % signal recovery of the reference material value.

4.14.3.3. Study 3- Onboard stability at 20-25℃ after reconstitution

The on-test and reference materials were tested in duplicate on the cobas e 411. The on-test material was reconstituted and stored in open vials for 6 hours at 20-25°C. The on-test signal recovery was calculated as percent of the reference value.

The acceptance criterion was 90 to 110 % signal recovery of the reference material value.

4.14.3.4. Real-time Stability

Since there was no change in formulation and target values to the Vitamin B12 II CalSet the realtime stability data (30 months at 2-8°C) of the current Vitamin B12 CalSet II is applicable.

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CLINICAL PERFORMANCE EVALUATION 5.

Clinical samples were collected at four sites in the United States in order to establish the reference range values for the Elecsys Vitamin B12 II assay. Reference ranges for apparently healthy males and apparently healthy females were determined using the median value and the 2.5th - 97.5th percentiles (pg/mL) as lower and upper limit of normal, respectively. The evaluation was done at one site in Germany (Roche Diagnostics GmbH, R&D department, Penzberg) with one reagent lot (MP) using one cobas e 411 analyzer. Samples were all native human serum samples measured in singleton, over 2 runs for 2 days.

6. CONCLUSIONS

The information provided in this Premarket Notification [510(k)] will support a determination of substantial equivalence for the Elecsys Vitamin B12 II Test System for the measurement of vitamin B12 in serum and plasma samples The data supports a safe, effective device which performs as well as or better than the predicate device.