(85 days)
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.
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) .
The document describes the Elecsys Vitamin B12 II Assay and Elecsys Vitamin B12 II CalSet, which are used for in vitro quantitative determination of Vitamin B12 in human serum and plasma. The device is intended for use in the diagnosis and treatment of anemias of gastrointestinal malabsorption.
Here's a breakdown of the requested information:
1. A table of acceptance criteria and the reported device performance
| Test Category | Acceptance Criteria | Reported Device Performance and Remarks |
|---|---|---|
| Precision | Elecsys Vitamin B12 II Assay (Candidate Device) | |
| Within-run (Repeatability) | ≤ 200 pg/mL: SD ≤ 14 pg/mL > 200 pg/mL: CV ≤ 7 % | Cobas e 411:HS 1 (176 pg/mL): 8.86 SD (5.0% CV)HS 2 (405 pg/mL): 13 SD (3.2% CV)HS 3 (960 pg/mL): 19.7 SD (2.1% CV)HS 4 (1230 pg/mL): 27.4 SD (2.2% CV)HS 5 (1940 pg/mL): 40.9 SD (2.1% CV)PCV0 (229 pg/mL): 8.96 SD (3.9% CV)PCV1 (447 pg/mL): 12.2 SD (2.7% CV)PCV2 (934 pg/mL): 20.2 SD (2.2% CV)All results met the acceptance criteria. |
| Intermediate Precision | ≤ 200 pg/mL: SD ≤ 24 pg/mL > 200 pg/mL: CV ≤ 12 % | Cobas e 411:HS 1 (176 pg/mL): 12.7 SD (7.2% CV)HS 2 (405 pg/mL): 17.5 SD (4.3% CV)HS 3 (960 pg/mL): 31.0 SD (3.2% CV)HS 4 (1230 pg/mL): 46.4 SD (3.8% CV)HS 5 (1940 pg/mL): 72.6 SD (3.7% CV)PCV0 (229 pg/mL): 12.4 SD (5.4% CV)PCV1 (447 pg/mL): 18.6 SD (4.2% CV)PCV2 (934 pg/mL): 38.4 SD (4.1% CV)All results met the acceptance criteria. |
| Limit of Blank (LoB) | LoB ≤ 50 pg/mL | Achieved. (Specific value not reported in the summary, but stated as meeting criterion.) |
| Limit of Detection (LoD) | LoD ≤ 100 pg/mL | Achieved. (Specific value not reported in the summary, but stated as meeting criterion.) |
| Limit of Quantitation (LoQ) | LoQ (Imprecision) ≤ 20 % at 150 pg/mL | Achieved. (Specific value not reported in the summary, but stated as meeting criterion.) |
| Linearity | ≤ 200 pg/mL: ± 20 pg/mL > 200 pg/mL: ± 10 % (for deviation to higher order polynomial regression) Significant level for deviation to higher order polynomial: 5% | Reported that "All results met the predefined acceptance criteria for linearity." and "The linearity results support a claimed measuring range." |
| Endogenous Interferences | < 200 pg/mL: ± 20 pg/mL of unspiked reference value > 200 pg/mL: ± 10% of unspiked reference value | Reported that "Predefined acceptance criterion was met." Claims included in the method sheet were set to the concentration without observed interference. |
| Exogenous Interferences - Drugs | ± 10% of the reference value (unspiked sample) | Reported that "No interference with the assay was found." for 16 commonly used pharmaceuticals. |
| Exogenous Interferences - Anticoagulants | Slope (Passing/Bablok): 0.9 – 1.1 Intercept (Passing/Bablok): < ± 100 pg/mL Pearson's r (linear regression): ≥ 0.95 Bias at 200 pg/mL: ± 10 % | Potential effects assessed by regression analysis. (Specific values not reported in summary, but implied to be acceptable based on overall conclusion of substantial equivalence). |
| Method Comparison (vs. Predicate) | Slope (Passing/Bablok): 1.00 ± 0.05 Correlation (linear regression) Pearson's r: > 0.95 Intercept (Passing/Bablok): < ± 30 pg/mL Bias at 200 pg/mL: ≤ ± 10 % | Reported for Candidate Device vs. Predicate:Slope (Passing/Bablok): 0.984Intercept (Passing/Bablok): -26.7Tau/r: 0.996Bias at 200 pg/mL: -15.0The values align with the criteria. |
| Reagent Stability (Recovery) | 80 - 120 % for PC Varia and human serum samples (for real-time stability) | Study 1, 2, and 3: Acceptance criterion for recovery was compared to day 0 value. (Implied to be met as no issues stated). Study 4 (real-time): Ongoing, data available for 0, 7, 10, 13, 19, 25 months; criterion is 80-120% recovery. |
| Sample Stability (Recovery) | ≤ 200 pg/mL: ± 28 pg/mL > 200 pg/mL: ± 14 % | Study 1, 2, and 3: Recovery was calculated as percent of the reference value. (Implied to be met as no issues stated). |
| Calibrator Reconstitution (Signal Recovery) | 90 to 110 % signal recovery of the reference material value | Achieved. (Specific values not reported in summary, but stated as meeting criterion.) |
| Calibrator Stability (-20°C, 2-8°C, Onboard) (Signal Recovery) | 90 to 110 % signal recovery of the reference material value | Achieved. (Specific values not reported in summary, but stated as meeting criterion.) |
2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)
- Precision: Not explicitly stated as a separate "test set" sample size but evaluated using "human serum samples" (HS 1-5) and "PreciControl Varia" (PCV0-2). The protocol involved testing 2 replicates per run, 2 runs per day for 21 days for precision.
- Limit of Blank (LoB): n = 60 LoB measurements (5 replicates, 2 runs per day on 2 instruments over 3 days) using a buffered human serum albumin matrix.
- Limit of Detection (LoD): n = 60 LoD measurements (5 samples, 2 runs per day on 2 instruments over 3 days) using 5 low-level human serum samples.
- Limit of Quantitation (LoQ): Minimum of seven human serum samples, analyzed in replicates of 5, one run per day over 5 days.
- Linearity: Serum samples.
- Analytical Specificity: Two human serum samples (single donors, native) spiked with potential cross-reactant compounds.
- Endogenous Interferences: Three human serum samples (single donors, native as well as spiked) for each interfering substance.
- Exogenous Interferences - Drugs: Two human serum samples (single donors, native) spiked with 16 pharmaceutical compounds.
- Exogenous Interferences - Anticoagulants: Minimum of 90 serum/plasma pairs per sample material (single donors, native as well as spiked).
- Method Comparison: 120 human serum samples (all single donors, native as well as spiked) covering the entire measuring range.
- Reference Range Study: 120 apparently healthy male & female subjects. Data provenance: USA for sample collection, evaluation done at one site in Germany. Samples were native human serum samples.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)
This is an in-vitro diagnostic (IVD) device for quantitative measurement of a biomarker (Vitamin B12). The "ground truth" for such devices is typically established through analytical methods and highly controlled reference materials, not through expert consensus on qualitative interpretation of images or clinical findings. The document does not describe the use of human experts to establish ground truth for this type of test.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
Not applicable. As an IVD device, the performance is assessed through analytical validation studies (precision, linearity, accuracy against reference methods/materials) rather than human expert adjudication of results.
5. 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 is an in-vitro diagnostic device for measuring Vitamin B12, not an imaging device or AI-assisted diagnostic tool that relies on human readers/interpreters.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
The device is a standalone in-vitro diagnostic assay (Elecsys Vitamin B12 II assay) intended to quantitatively measure Vitamin B12. Its performance is evaluated analytically, separate from human interpretation of the results for direct diagnostic decision-making, though the results contribute to a physician's overall diagnosis and treatment plan. The studies described (precision, linearity, LoB/LoD/LoQ, interference, method comparison) are all "standalone" evaluations of the device's analytical performance.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The ground truth for the analytical and method comparison studies is established based on:
- Reference methods/materials (e.g., specific calibrators, spiked samples with known concentrations, comparison to the legally marketed predicate device).
- Defined analytical standards and statistical methods (e.g., CLSI guidelines).
- The "value assignment" for calibrators is a key part of establishing the reference for the assay.
8. The sample size for the training set
This document does not describe a "training set" in the context of machine learning. The studies described are validation and verification studies for an analytical assay. The "Elecsys Vitamin B12 II CalSet" serves as a calibrator for the assay, analogous to the concept of a "calibration set" for an analytical instrument, but not a training set for an AI/ML algorithm.
9. How the ground truth for the training set was established
Not applicable as there is no "training set" in the machine learning sense. However, for the calibrator (Elecsys Vitamin B12 II CalSet), its "target values" are assigned to obtain the best fit with the Master Calibration Curve, using multiple analyzers (at least 3 cobas e 411 and at least 3 cobas e 601/MODULAR ANALYTICS E170 analyzers) and multiple reagent lots. The assigned value for each calibrator is the mean value obtained over at least six runs on at least three analyzers. PreciControl Varia is also used to monitor accuracy and precision of analytes, and acceptance criteria for these controls must be met to release the assigned values for the CalSet. This process establishes the "ground truth" for the calibrators.
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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.
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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.
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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:
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"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 RoadIndianapolis, IN 46250 |
| Contact | Kelli TurnerPhone: (317) 521-4515FAX: (317) 521-2324Email: kelli.turner@roche.com |
| Date Prepared | 06/29/2015 |
| Proprietary Name | 1. Elecsys Vitamin B12 II assay,2. Elecsys Vitamin B12 II CalSet |
| Common Name | 1. Vitamin B12 II assay2. Vitamin B12 II CalSet |
| Classification Name | 1. Radioassay, Vitamin B122. Secondary, calibrator |
| Product Codes | 1. CDD, 862.18102. JIT, 862.1150 |
| Predicate Devices | Elecsys Vitamin B12 Assay (K060755) |
| Establishment Registration | Roche Diagnostics GmbH in Mannheim, Germany, is 9610126Roche Diagnostics GmbH in Penzberg, Germany, is 9610529Roche 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 forUse | Binding assay for the in vitro quantitativedetermination of vitamin B12 in human serumand plasma.The electrochemiluminescence immunoassay"ECLIA" is intended for use on Elecsys andcobas e immunoassay analyzers.A Vitamin B12 test system is a device intendedto measure Vitamin B12 in serum, plasma.Measurements obtained by this device areused in the diagnosis and treatment of anemiasof gastrointestinal malabsorption. | Binding assay for the in vitro quantitativedetermination of vitamin B12 in human serumand plasma. Measurements obtained by thisdevice are used in the diagnosis and treatmentof anemias of gastrointestinal malabsorption.The electrochemiluminescence immunoassay"ECLIA" is intended for use on Elecsys andcobas e immunoassay analyzers. |
| Assay Protocol | The Elecsys Vitamin B12 assay employs acompetitive test principle usingintrinsic factor specific for vitamin B12. VitaminB12 in the sample competes with the addedvitamin B12 labeled with biotin for the bindingsites on the ruthenium- labeled intrinsic factorcomplex. | Same. |
| DetectionProtocol | Electrochemiluminescent Assay | Same. |
| Applications | 27-minute application | Same. |
| InstrumentPlatform | Elecsys and cobas e immunoassay analyzers. | Same. (submission for cobas e 411) |
| Sample Volume | 15 µL | Same. |
| SampleType | 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 IIAssay |
|---|---|---|
| CalibrationInterval | Calibration must be performed once perreagent lot using fresh reagent (i.e. not morethan 24 hours since the reagent kit wasregistered on the analyzer). Renewedcalibration is recommended as follows:• After 1 month (28 days) when using thesame reagent lot.• After 7 days (when using the samereagent kit on the analyzer).• As required: e.g. quality control findingsoutside the defined limits | Same. |
| Controls | Elecsys PreciControl Varia | Same |
| Traceability /Standardization | Standardized against the Elecsys Vitamin B12assay (K973702) | Standardized against the Elecsys Vitamin B12assay (K060755).Accuracy to WHO Standard 03/178 |
| ReagentStability | Unopened: 2-8°C - Up to the stated expirationdate.After Opening at 2-8°C - 12 weeksOn the Analyzers – 5 weeks | Unopened: 2-8°C - Up to the stated expirationdate.After Opening at 2-8°C - 12 weeksOn the Analyzers - 5 weeks onboard or 60days when stored alternatively in therefrigerator and on the analyzer, with the totaltime onboard on the analyzer not exceeding 10x 8 hours |
| MeasuringRange | 30.0 - 2000 pg/mL | 150- 2000 pg/mL |
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| Feature | Predicate Device: Elecsys Vitamin B12(K060755) | Candidate Device: Elecsys Vitamin B12 II Assay | ||||||
|---|---|---|---|---|---|---|---|---|
| Precision | cobas e 411:Within-run | cobas e 411:Within-run (will be labeled Repeatability) | ||||||
| Sample | Mean (pg/mL) | SD | CV | Sample | Mean (pg/mL) | SD | CV | |
| HS 1 | 192 | 11.3 | 5.9% | HS 1 | 176 | 8.86 | 5.0% | |
| HS 2 | 358 | 20.1 | 5.6% | HS 2 | 405 | 13 | 3.2% | |
| HS 3 | 864 | 29.1 | 3.3% | HS 3 | 960 | 19.7 | 2.1% | |
| HS 4 | 1625 | 62.1 | 3.8% | HS 4 | 1230 | 27.4 | 2.2% | |
| PCV0 | 221 | 10.2 | 4.6% | HS 5 | 1940 | 40.9 | 2.1% | |
| PCV1 | 467 | 18.5 | 4.0% | PCV0 | 229 | 8.96 | 3.9% | |
| PCV2 | 958 | 31.1 | 3.3% | PCV1 | 447 | 12.2 | 2.7% | |
| PCV2 | 934 | 20.2 | 2.2% | |||||
| Total | Total (will be labeled Intermediate precision) | |||||||
| Sample | Mean (pg/mL) | SD | CV | Sample | Mean (pg/mL) | SD | CV | |
| HS 1 | 192 | 19.8 | 10.3% | HS 1 | 176 | 12.7 | 7.2% | |
| HS 2 | 358 | 27.7 | 7.7% | HS 2 | 405 | 17.5 | 4.3% | |
| HS 3 | 864 | 40.1 | 4.5% | HS 3 | 960 | 31.0 | 3.2% | |
| HS 4 | 1625 | 65.1 | 4.0% | HS 4 | 1230 | 46.4 | 3.8% | |
| PCV0 | 221 | 10.2 | 4.6% | HS 5 | 1940 | 72.6 | 3.7% | |
| PCV1 | 467 | 18.5 | 4.0% | PCV0 | 229 | 12.4 | 5.4% | |
| PCV2 | 958 | 31.1 | 3.3% | PCV1 | 447 | 18.6 | 4.2% | |
| PCV2 | 934 | 38.4 | 4.1% | |||||
| AnalyticalSensitivity | Lower detection limit = 30.0 pg/mL | Limit of Blank (LoB): = 50 pg/mL Limitof Detection (LoD): = 100 ng/mLLimit of Quantitation (LoQ): = 150 pg/mL | ||||||
| AnalyticalSpecificity | Cross reactant | concentrationtested(ng/mL) | Highest cross-reactivityobserved (%) | Cross reactant | concentrationtested(ng/mL) | Highest cross-reactivityobserved (%) | ||
| Cobinamide | 200 | 0.024 | Cobinamide | 210 | 0.003 | |||
| Linearity | 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 IIAssay |
|---|---|---|
| Limitations | The assay is unaffected by:• Bilirubin < 65 mg/dL• Hemolysis < 1.0 g/dL• Lipemia < 1500 mg/dL• Biotin < 50 ng/mL• Rheumatoid factors < 1500 IU/mL• In vitro tests were performed on54 commonly used pharmaceuticals. Nointerference with the assay was found.• In rare cases, interference due toextremely high titers of antibodies toanalyte-specific antibodies, streptavidinor ruthenium can occur. These effectsare minimized by suitable test design.For diagnostic purposes, the results shouldalways be assessed in conjunction with RBCfolate, the patient's medical history, clinicalexamination and other findings. | The assay is unaffected by:• Bilirubin ≤ 65 mg/dL• Hemolysis ≤ 0.1 g/dL• Lipemia ≤ 1500 mg/dL• Biotin ≤ 50 ng/mL• Rheumatoid factors < 1500 IU/mL• IgG ≤ 28 g/dL• IgM ≤ 10 g/dL• IgA ≤ 16 g/dL• In vitro tests were performed on16 commonly used pharmaceuticals. Nointerference with the assay was found.• In rare cases, interference due toextremely high titers of antibodies toanalyte-specific antibodies, streptavidinor ruthenium can occur. These effectsare minimized by suitable test design.• Because intrinsic factor is typically usedas the binding protein in serum vitaminB12 assays, anti-intrinsic factorantibodies (which are common inpernicious anemias) can lead toelevated vitamin B12 measurementvalues.• The Elecsys Vitamin B12 II assay isdesigned to avoid interference due toanti-intrinsic factor antibodies.For diagnostic purposes, the results shouldalways be assessed in conjunction with thepatient's medical history, clinical examinationand other findings.Note: the presence of immunoglobulin-vitaminB12 complexes may cause unexpectedly highvalues of vitamin B12. |
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| Feature | Predicate Device: Elecsys Vitamin B12(K060755) | Candidate Device: Elecsys Vitamin B12 II Assay | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Referencerange study | Country | (N) | Median | 2.5th-97.5th percentile | Subject cohort | (N) | Median | 2.5th-97.5th percentile | ||||
| USA | 178 | pg/mL | Pmol/L | pg/mL | pmol/L | Apparently healthy male & female | 120 | pg/mL | pmol/L | pg/mL | pmol/L | |
| 463 | 342 | 211-946 | 156-698 | 443 | 327 | 232-1245 | 171-919 | |||||
| MethodComparison | n = 120 | Passing/Bablok | Linear regression | |||||||||
| Min = 156 pg/mL | ||||||||||||
| Max = 1753 pg/mL | ||||||||||||
| Slope | 0.950 | 0.984 | ||||||||||
| Intercept | -9.83 | -26.7 | ||||||||||
| Tau/r | 0.941 | 0.996 | ||||||||||
| Bias at 200 pg/mL | -9.88 | -15.0 |
Table 2: Assay Comparison: Labeled Performance Characteristics
HS= Human Serum
PCV0=PreciControl Varia level 0
PCV1=PreciControl Varia level 1
PCV2=PreciControl Varia level 2
Table 3: CalSet Comparison
| Characteristic | Predicate device: Elecsys Vitamin B12CalSet II | Candidate device: Elecsys Vitamin B12 II |
|---|---|---|
| Intended Use | Vitamin B12 CalSet II is used for calibrating thequantitative Elecsys Vitamin B12 assay on theElecsys and cobas e immunoassay analyzers. | Vitamin B12 II CalSet is used for calibrating thequantitative Elecsys Vitamin B12 II assay onthe Elecsys and cobas e immunoassayanalyzers. |
| Analyte | Vitamin B12 | Same |
| Matrix | Human serum matrix | Same |
| Levels | Two | Same |
| Target Ranges | Cal 1: 250 pg/mLCal 2: 1500 pg/mL | Same. |
| Format | Lyophilized | Same |
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| Characteristic | Predicate device: Elecsys Vitamin B12CalSet II | Candidate device: Elecsys Vitamin B12 II |
|---|---|---|
| Handling | Carefully dissolve the contents of one bottle byadding exactly 1.0 mL of distilled or deionizedwater and allow to stand closed for 15 minutesto reconstitute. Mix carefully, avoiding foamformation.Transfer aliquots of the reconstituted calibratorsinto empty labeled snap- cap bottles(CalSet Vials). Attach the supplied labels to theadditional bottles. Store the aliquotsimmediately at - 20 °C.Perform only one calibration procedureper aliquot. | Same. |
4. NON-CLINICAL PERFORMANCE EVALUATION
Non-clinical performance evaluations for the Elecsys Vitamin B12 II executed with the study briefly summarized.
4.1. Precision
The precision results were obtained using serum samples evaluated on the cobas e 411 Immunoassay Analyzer. Within-run precision (repeatability) and total imprecision (intermediate precision) were determined according to CLSI Guideline EP5-A2. The protocol included testing 2 replicates per run, 2 runs per day for 21 days.
Specifications:
| Repeatability | Intermediate precision | |
|---|---|---|
| ≤ 200 pg/mL | SD ≤ 14 pg/mL | SD ≤ 24 pg/mL |
| > 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: ± 20 pg/mL of unspiked reference value
-
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): <±100 pg/mL .
- coefficient of correlation Pearson's r (linear regression): ≥ 0.95 .
- Bias at 200 pg/mL: ± 10 % .
4.11. Method Comparison
A method comparison was performed using the current Elecsys Vitamin B12 assay (cleared under K060755) as predicate device.
A total of 120 human serum samples (all single donors, native as well as spiked) were measured in singleton covering the entire measuring range.
The study was performed on the cobas e 411 analyzer over 3 runs using the current Elecsys Vitamin B12 assay (X) and the updated Elecsys Vitamin B12 II assay (Y). Regression analysis was performed.
Acceptance criteria:
- 1.00 ± 0.05 . Slope (Passing/Bablok):
- Correlation (linear regression) Pearson's r: . > 0.95
- Intercept (Passing/Bablok): < ± 30 pg/mL .
- Bias at 200 pg/mL: ≤± 10 % .
4.12. Reagent Stability
To test reagent stability, four studies were executed with three studies completed.
4.12.1. Study 1. Reagent stability refrigerator/onboard (60 days)
Reagent stability refrigerator / onboard was tested on one cobas e 411 Immunoassay Analyzer.
A fresh reagent rackpack (kit) was placed on the analyzer and calibrated. Reference values for the five human serum (HS) samples and three controls were determined in duplicate. After
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measurement the rackpack was removed from the analyzer and kept at 2-8 °C for 64 days. During this period of time, the kit was stressed alternately at 20°C + 3°C (on-board condition) 10 times for 8 hours (80 hours in total). On day 8, 15, 29, 50 and day 64, the same samples were measured with this reagent kit kept under alternating storage conditions (refrigerator /on-board) using the calibration curves established on day 0, 8, 22, 43 and 57, respectively. Acceptance criterion for recovery was compared to day 0 value.
4.12.2. Study 2. Reagent stability after first opening at 2-8°C (84 days)
Reagent stability after first opening for the Elecsys Vitamin B12 II assay was tested on one cobas e 411 Immunoassay Analyzer.
A fresh reagent rackpack was placed on the analyzer and calibrated. Reference values for the five human serum (HS) samples and three controls were determined in duplicate. After measurement the kit was removed from the analyzer and kept at 2-8 °C up to 92 days. After 36, 64 and 92 days the kit was placed on the analyzer again, calibrated and the test samples were determined. Acceptance criterion for recovery was compared to day 0 value.
4.12.3. Study 3. On board reagent stability (35 days)
Reagent On-board Stability and Calibration Stability for the Elecsys Vitamin B12 II assay was tested on one cobas e 411 Immunoassay Analyzer. A fresh Reagent Rack-Pack was placed on the analyzer and calibrated. Reference values for the five human serum (HS) samples and three controls were measured in duplicate on day 0. On day 8, 22 and day 36, the same samples were measured with the same reagent kit kept at 20°C + 3°C (on-board condition) using the calibration curves established on day 0, 15 and 29, respectively. Acceptance criterion for recovery was compared to day 0 value.
4.12.4. Study 4: A real-time stability study is ongoing to support shelf-life stability claim.
In the ongoing real-time stability study, the Elecsys Vitamin B12 II reagent is stored at 2-8°C. The stored assay reagents are tested at time point T=0 (after manufacturing) and at specified intervals over the shelf life of the device up to the planned shelf life plus one month. Testing will be performed using PreciControl Varia 3. Level 0. 1 and 2 (lyopylized, stored at -20℃) and in addition three human serum samples (stored at -80°C).
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Data for the time-points at 0, 7, 10, 13, 19 and 25 months tested in duplicate will be available. The average on-test recovery value will be calculated as percent recovery compared to the reference value:
- assigned value for PreciControl Varia 3, Level 0, 1 and 2 .
- value measured at T=0 for the human serum samples .
The acceptance criterion is recovery of 80 -120 % for PC Varia and the human serum samples.
4.13. Sample Stability
To test sample stability, three studies were completed. Because these studies are not analyzer dependent, and the assay is a global product currently available rest of world (ROW), the studies were executed on the cobas e 601. Study results can be applied to the cobas e 411 since sample stability is independent of the analyzer. For each study, the samples used were all single donors (native, spiked and diluted) and the following acceptance criteria was applied.
Acceptance criteria for recovery compared to the reference value:
- ≤ 200 pg/mL: ± 28 pg/mL .
-
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.
§ 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.