The VB12 method is an in vitro diagnostic test for the quantitative measurement of Vitamin B12 (B12) in human serum and plasma on the Dimension® EXL™ with LM integrated chemistry system. Measurements of Vitamin B12 may be used in the diagnosis of vitamin B12 deficiency.

The LOCI ANEMIA CAL is an in vitro diagnostic product for the calibration of the LOCI FOLA and LOCI VB12 assays on the Dimension® EXL™ with LM integrated chemistry system.

Device Description

The vitamin B12 method is a homogeneous, competitive chemiluminescent immunoassay based on LOCI® technology. LOCI® reagents include two synthetic bead reagents and biotinylated intrinsic factor (IF). The first bead reagent (Chemibead) is coated with a B12 derivative and contains a chemiluminescent dve. The second bead reagent (Sensibead) is coated with streptavidin and contains photosensitive dve. The patient sample is pretreated with sodium hydroxide (NaOH) and dithioerythritol (DTE) to release the serum B12 from its carrier proteins. Potassium cyanide (KCN) is added to convert all the forms of B12 into a single, cyanocobalamin form, and dicyanocobinamide is added to keep the B12 from rebinding with the carrier proteins. After the sample pretreatment, the biotinylated IF and chemibead reagents are added sequentially to the reaction vessel. Vitamin B12 from the sample competes with the B12-chemibead for a limited amount of biotinylated IF. Sensibead reagent is then added and binds to the biotin to form bead pair immunocomplexes. Illumination of the complex at 680 nm generates singlet oxygen from the Sensibeads which diffuses to the Chemibeads triggering a chemiluminescent reaction. The resulting signal is measured at 612 nm and is an inverse function of the concentration of vitamin B12 in the sample.

LOCI ANEMIA CAL is a multi-analyte liquid, frozen product containing Folate and Vitamin B12. All levels are prepared in a bovine serum albumin base. The kit consists of ten vials, two each of five levels containing 2 mL per vial.

AI/ML Overview

The provided 510(k) summary focuses on establishing substantial equivalence for an in vitro diagnostic (IVD) device, specifically the LOCI Vitamin B12 Flex® reagent cartridge and the LOCI Anemia Calibrator, to predicate devices. This type of filing differs from those for AI/ML-enabled devices, which often involve comparative effectiveness studies with human readers.

As a result, the information required for a typical AI/ML-driven device's acceptance criteria and study design (such as sample size for test sets, data provenance, number of experts for ground truth, adjudication methods, MRMC studies, standalone performance metrics, training set details) is not present in this document. The study described here is primarily a method comparison and reproducibility study for an IVD assay, not an AI/ML algorithm that predicts or interprets clinical data for human-in-the-loop use.

However, I can extract the acceptance criteria and reported performance relevant to this IVD device's analytical validation.

Acceptance Criteria and Reported Device Performance (Analytical Validation)

Since this is an IVD device submission establishing substantial equivalence to a predicate, the "acceptance criteria" are implicitly tied to demonstrating comparable analytical performance to the predicate device and meeting analytical precision expectations.

Acceptance Criteria Category	Specific Metric (Implicit)	Acceptance Criteria (Implicit)	Reported Device Performance (LOCI VB12 Flex® reagent cartridge)	Study Type
Reproducibility / Precision	Repeatability (SD, %CV)	Acceptable precision for clinical IVD	Bio-Rad Liquichek® Immunoassay ControlL1: 13.2 SD, 4.6%CVL2: 11.7 SD, 2.3%CVL3: 15.9 SD, 2.5%CV Serum Pool 1: 10.1 SD, 5.6%CVLi Heparin Plasma Pool: 13.1 SD, 2.8%CVSerum Pool 2: 24.9 SD, 2.5%CVSerum Pool 3: 27.4 SD, 1.6%CV	Reproducibility
	Within-Lab Precision (SD, %CV)	Acceptable precision for clinical IVD	Bio-Rad Liquichek® Immunoassay ControlL1: 15.0 SD, 5.2%CVL2: 18.2 SD, 3.7%CVL3: 21.5 SD, 3.3%CV Serum Pool 1: 11.6 SD, 6.5%CVLi Heparin Plasma Pool: 15.8 SD, 3.4%CVSerum Pool 2: 27.7 SD, 2.8%CVSerum Pool 3: 35.2 SD, 2.0%CV	Reproducibility
Method Comparison (vs. Predicate)	Bias (Intercept)	Intercept and Slope for Passing-Bablok regression should show good agreement (close to 0 and 1, respectively) for substantial equivalence.	Intercept: -6.37 pg/mL (95% CI: -12.90 to 0.36 pg/mL)	Method Comparison
	Bias (Slope)	Intercept and Slope for Passing-Bablok regression should show good agreement (close to 0 and 1, respectively) for substantial equivalence.	Slope: 1.03 (95% CI: 1.02 to 1.04)	Method Comparison
	Correlation Coefficient (r)	High correlation (e.g., >0.95 or >0.975 often seen as a benchmark for good agreement)	0.997 (using least squares regression)	Method Comparison
Sample Type Equivalence	Passing-Bablok analysis vs. Serum (Slope)	Slopes should be close to 1 (e.g., 0.98 - 1.02) to indicate equivalence.	Lithium Heparin Plasma: 1.00Sodium Heparin Plasma: 1.02EDTA Plasma: 1.00SST Tubes: 1.01	Sample Comparison
	Passing-Bablok analysis vs. Serum (Intercept)	Intercepts should be close to 0 to indicate equivalence.	Lithium Heparin Plasma: -4.33 pg/mLSodium Heparin Plasma: -10.73 pg/mLEDTA Plasma: -7.26 pg/mLSST Tubes: -2.31 pg/mL	Sample Comparison

Study Details (as inferable from the document):

Sample size used for the test set and the data provenance:
- Reproducibility Study: Not explicitly stated as a "test set" in the AI/ML sense, but for the precision study, multiple runs over 20 days were performed for each material (3 Bio-Rad controls, 4 serum/plasma pools). Each test level was analyzed twice per day for 20 days.
- Sample Type Equivalency Study: 78 matched serum, lithium heparin plasma, sodium heparin, and EDTA plasma samples. 71 were native, 6 spiked, 1 diluted. Data provenance is not specified (e.g., country of origin, retrospective/prospective).
- Method Comparison Study: 166 native human serum samples. These ranged from 60 - 1966 pg/mL (predicate) and 86 - 1901 pg/mL (new device). 35 samples were positive for Intrinsic Factor Blocking Antibody (IFBA). Data provenance is not specified.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts: Not applicable. For IVD devices, "ground truth" typically refers to the reference method or assigned values, not expert interpretation of images or clinical scenarios. In the method comparison, the predicate device's measurements served as a reference for comparison. Calibrator values were assigned from a master pool with traceability to USP Grade Vitamin B12.
Adjudication method (e.g., 2+1, 3+1, none) for the test set: Not applicable, as this is an analytical performance study for an IVD, not a diagnostic interpretation study.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance: Not applicable. This submission is for an IVD assay, not an AI/ML-enabled diagnostic imaging or clinical decision support system that involves human readers.
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done: The studies described assess the standalone analytical performance of the IVD assay without human interpretation of results beyond standard laboratory practices.
The type of ground truth used:
- For the method comparison, the predicate device's measurements (Dimension Vista® VB12 method, K121994) served as the comparative reference.
- For the calibrator, values are assigned from a master pool which has traceability to USP Grade Vitamin B12.
- For precision, the established analytical performance characteristics for laboratory controls and serum/plasma pools serve as benchmarks.
The sample size for the training set: Not applicable. This is an IVD assay, not an AI/ML device requiring a training set in that context. The device's calibration involves specific calibrator materials with assigned values.
How the ground truth for the training set was established: Not applicable. The "ground truth" for the IVD's calibration is established via a master pool traceable to USP Grade Vitamin B12.

Summary

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510(k) Summary

This summary of 510(k) safety and effectiveness information is submitted in accordance with the requirements of SMDA 1990 and 21 CFR §807.92.

K133512

The assigned 510(k) number is:

1. Manufacturer's Name, Address, Telephone, and Contact Person, Date of Preparation

Manufacturer:

Contact Information:

Siemens Healthcare Diagnostics Inc. P.O. Box 6101 Newark, DE 19714

Siemens Healthcare Diagnostics Inc. P.O. Box 6101 Newark, DE 19714 Attn: A. Kathleen Ennis Regulatory Affairs Manager Tel: 302-631-9352 FAX # 302-631-6299

Date of Preparation:

December 30, 2013

2. Device Name

Proprietary Name

. LOCI Vitamin B12 Flex® reagent cartridge
LOCI Anemia Calibrator .
Common Name
- Vitamin B12 Assay .
- Calibrator ●
FDA Classification
- Radioassay Vitamin B12 code CDD ●
- Calibrator Multianalyte - code JIX .

3. Identification of the Predicate Device

. Dimension Vista® Vitamin B12 Flex® reagent cartridge (VB12) K121994
Dimension Vista® LOCI 4 CAL K121994 .

FDA Guidance Document(s):

"Bundling Multiple Devices or Multiple Indications in a Single ● Submission" - 11/26/2003

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Device Description(s): 4.

LOCI VB12 Assav

Wellsa	Form	Ingredient	Concentrationb	Source
1	Liquid	VB12 Sensibead reagent	360 µg/mL	Recombinant E. coli
2	Liquid	VB12-Chemibead reagent	200 µg/mL
3	empty
4	Liquid	Biotinylated IF	3 ng/mL	porcine
		Dicyanocobinimide	60 ng/mL
5	Tablet	Dithioerythritol (DTE)	12.5 mg/mL
6	Liquid	Sodium Hydroxide (NaOH)	0.75 N
		Potassium Cyanide (KCN)	3 mM
7-8	empty

The LOCI VB12 method reagents are packaged in an eight-well Flex® reagent cartridge as follows:

a. Wells are numbered consecutively from the wide end of the cartridge.

b. Nominal value per well in a cartridge.

Each cartridge contains a sufficient to perform 20 tests. Four (4) Flex® reagent cartridges are provided in each carton.

LOCI Anemia Calibrator

LOCI ANEMIA CAL is a multi-analyte liquid, frozen product containing Folate and Vitamin B12. All levels are prepared in a bovine serum albumin base.

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The kit consists of ten vials, two each of five levels containing 2 mL per vial. Description of the manufacturing, value assignment and stability testing processes are provided.

Values are assigned to each lot of calibrator from a master pool using the Dimension® EXL with LM integrated chemistry system. The master pool is a frozen liquid, five level material, with the same composition as the calibrator with traceability to USP Grade Vitamin B12.

The assigned to LOCI Anemia Calibrator are stable for unopened frozen product until the expiration date on the label. Unopened (Thawed) product is stable for 30 days when stored at 2-8°C. Once the product is opened (cap removed), assigned values are stable for 30 days when recapped immediately after use and stored at 2-8°C. Shelf life and in use stability have been established by Siemens Healthcare through real time testing at the temperatures specified.

5. Device Intended Use:

LOCI VB12 Assay

LOCI Anemia Calibrator

The LOCI ANEMIA CAL is an in vitro diagnostic product for the calibration of the LOCI FOLA and LOCI VB12 assays on the Dimension® EXL™ with LM integrated chemistry system.

6. Medical device to which equivalence is claimed:

Substantial Equivalence:

The LOCI Vitamin B12 Flex® reagent cartridge (RF642) is substantially equivalent to the Dimension Vista® Vitamin B12 method (k#121994). The LOCI Anemia Calibrator (RC640) is substantially equivalent to the LOCI 4 Calibrator, cat.# KC640A (k#121994).

Comparison to Predicate Device:

The proposed Siemens Healthcare Diagnostics LOCI VB12 method and the predicate Dimension Vista® VB12 method (K121994) are both in vitro diagnostic immunoassays intended for the quantitative measurement of vitamin B12 in serum and plasma.

The Siemens Healthcare Diagnostics LOCI Anemia Calibrator, cat, # RC640 and the predicate Siemens LOCI 4 Calibrator, cat. # KC640A (K121994) are both used for the calibration of Folate and Vitamin B12 methods.

A comparison summary of the features of the products is included in the following table.

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LOCI VB12 Assay:

Item	DeviceLOCI Vitamin B12 Flex®reagent cartridge	PredicateDimension Vista® VitaminB12 Flex® reagent cartridge(K121994)
Similarities
Intended Use	in vitro diagnostic test for thequantitative measurement ofvitamin B12 in human serumand plasma	in vitro diagnostic test for thequantitative measurement ofvitamin B12 in human serumand plasma
Sample Types	Serum and Plasma	Serum and Plasma
Measurementmethod	Chemiluminescent:Homogenous sandwichimmunoassay based on LOCI®technology	Chemiluminescent:Homogenous sandwichimmunoassay based on LOCI®technology
Sample Size	12 uL	12 uL
Differences
Instrument	The Dimension® EXL™ withLM System	The Dimension Vista® System.
Measuring Range	80 - 2000 pg/mL	60 - 2000 pg/mL

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LOCI Anemia Calibrator:

Item	DeviceLOCI Anemia Calibrator(ANEM CAL)	PredicateLOCI 4 Calibrator(K121994)
Similarities
Intended Use	The LOCI ANEM CAL is an invitro diagnostic product for thecalibration of the LOCI FOLAand LOCI VB12 assays on theDimension® EXLT™ with LMSystem.	The LOCI 4 CAL is an in vitrodiagnostic product for thecalibration of the LOCI Ferritin(FERR), LOCI Folate (FOL) andLOCI Vitamin B12 (VB12)methods on the DimensionVista® System.
Traceability
Folate	United States PharmacopeiaGrade Folic Acid	United States PharmacopeiaGrade Folic Acid
Vitamin B12	United States PharmacopeiaGrade vitamin B12	United States PharmacopeiaGrade vitamin B12
Form	Frozen Liquid	Frozen Liquid
TargetConcentrationsVB12	Level 1 : 45 pg/mLLevel 2 200 pg/mLLevel 3 500 pg/mLLevel 4 1000 pg/mLLevel 5: 2200 pg/mL	Level A: 45 pg/mLLevel B: 200 pg/mLLevel C: 500 pg/mLLevel D: 1000 pg/mLLevel E: 2200 pg/mL
Folate	Level 1: 0 ng/mLLevel 2: 2.5 ng/mLLevel 3: 5.0 ng/mLLevel 4: 10.0 ng/mLLevel 5: 21.0 ng/mL	Level A: 0 ng/mLLevel B: 2.5 ng/mLLevel C: 5.0 ng/mLLevel D: 10.0 ng/mLLevel E: 21.0 ng/mL
Differences
Matrix	bovine serum albumin base inlevels 1 - 5	HEPES buffer Level Abovine serum base (BSA) inlevels B - E
Constituents	LOCI ANEM CAL is assigned forvitamin B12 and Folate.	LOCI 4 CAL is assigned forvitamin B12, Folate and Ferritin

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Comments on Substantial Equivalence:

Method

The Siemens Healthcare Diagnostics Dimension® LOCI VB12 method and the predicate Siemens Healthcare Diagnostics Dimension Vista® LOCI VB12 method (K121994) are both in vitro diagnostic immunoassays intended for the measurement of vitamin B12 in serum and plasma.

Reproducibility testing was conducted for the Dimension® LOCI VB12 method in accordance with the CLSI/NCCLS Approved Guideline for User Evaluation of Precision Performance of Clinical Chemistry Devices EP5-A2. For each test level, a single test from two independent cups was analyzed twice per day for 20 days. The repeatability and within-lab standard deviations were calculated by the analysis of variance method.

Material	Meanpg/mL	Repeatability		Within-Lab Precision
		SD	%CV	SD	%CV
Bio-Rad Liquichek® ImmunoassayControl*
L1	290	13.2	4.6	15.0	5.2
L2	498	11.7	2.3	18.2	3.7
L3	645	15.9	2.5	21.5	3.3
Serum Pool 1	180	10.1	5.6	11.6	6.5
Li Heparin PlasmaPool	467	13.1	2.8	15.8	3.4
Serum Pool 2	978	24.9	2.5	27.7	2.8
Serum Pool 3	1733	27.4	1.6	35.2	2.0

Typical precision observed for the Dimension® LOCI VB12 method is summarized below:

*BioRad Liquichek™ Immunoassay Plus Controls, BioRad Laboratories, Irvine, CA

A total of seventy-eight (78) matched serum, lithium heparin plasma, sodium heparin and EDTA plasma samples were analyzed on the Dimension ® EXL™ System. Of these 71 were native samples, 6 were spiked and one was diluted. Each plasma type was analyzed versus serum. Serum Separator tubes (SST) were analyzed in the same study. The Passing-Bablok regression analysis was used to analyze the data. The results are as follows:

Sample Type (vs Serum)n = 78	Slope	95%ConfidenceInterval	Interceptpg/mL	95%ConfidenceInterval pg/mL
Lithium Heparin Plasma	1.00	0.99 - 1.02	-4.33	-13.4 to +2.5
Sodium Heparin Plasma	1.02	0.99 - 1.04	-10.73	-20.2 to -0.4
EDTA Plasma	1.00	0.98 - 1.02	-7.26	-15.8 to +3.5
SST Tubes	1.01	0.99 - 1.03	-2.31	12.0 to +6.3

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A method comparison between the LOCI VB12 assay on the Dimension® EXL™ with LM System and the predicate, Dimension Vista® VB12 method (K121994), was performed with 166 native human serum samples across the proposed assay range. The samples ranged from 60 - 1966 pg/mL with the Dimension Vista® VB12 method, and, 86 - 1901pq/mL with the LOCI® VB12 assay on the Dimension® EXL™ with LM integrated chemistry system. These samples included thirty-five (35) samples that were positive for Intrinsic Factor Blocking Antibody (IFBA) with titers ranging from 15 - 164 IU/mL.

Passing-Bablok regression analysis of the results yielded the following:

Passing & Bablok Regression
	Bias	95% CI
Intercept(pg/mL)	-6.37	-12.90 to 0.36
Slope	1.03	1.02 to 1.04

The correlation coefficient, using least squares regression, for this data set (r) is 0.997.

Conclusion:

The Siemens Healthcare Diagnostics LOCI VB12 method and the predicate Siemens Healthcare Diagnostics Dimension Vista® VB12 method (K121994) are substantially equivalent based on their intended use and performance characteristics as described above. The calibrator products, the Siemens Healthcare Diagnostics LOCI Anemia Calibrator and the predicate Dimension Vista® LOCI 4 Calibrator (K121994) are also substantially equivalent in its design and intended use with their respective assay systems.

Anna Marie K. Ennis Requlatory Affairs Manager December 30 2013

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Image /page/7/Picture/0 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features a stylized depiction of an eagle or bird-like figure with three curved lines representing its wings or body. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the emblem.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

January 2, 2014

SIEMENS HEALTHCARE DIAGNOSTICS ANNA MARIE KATHLEEN ENNIS P.O. Box 6101 500 GBC DRIVE M/S 514 NEWARK DE 19714-6101

Re: K133512

Trade/Device Name: LOCI Vitamin B12 Flex® Reagent Cartridge and LOCI Anemia Calibrator

Regulation Number: 21 CFR 862.1810 Regulation Name: Vitamin B12 test system Regulatory Class: II Product Code: CDD, JIX Dated: November 5, 2013 Received: November 15, 2013

Dear Ms. Ennis:

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

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Page 2-Ms. Ennis

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 Small Manufacturers, International and Consumer Assistance 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" (21CFR 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 Small Manufacturers, International and Consumer Assistance 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.

Carol C. Benson -S for

Courtney H. Lias, Ph.D. 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 (if known): K133512

Device Name: LOCI vitamin B12 Flex® Reagent Cartridge and LOCI Anemia Calibrator

Indications for Use:

The LOCI ANEMIA CAL is an in vitro diagnostic product for the calibration of the LOCI FOLA and LOCI VB12 assays on the Dimension® EXL™ with LM integrated chemistry system.

Prescription Use X (21 CFR Part 801 Subpart D) And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR)

YungFDAban-S

Division Sign-Off Office of In Vitro Diagnostics and Radiological Health

510(k) K133512

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.