(27 days)
The LIAISON® 25 TOTAL-D assay is a chemiluminescent immunoassay (CLIA) intended for the quantitative determination of 25-hydroxyvitamin D and other hydroxylated vitamin D metabolites in human serum, EDTA and Lithium heparin plasma. The LIAISON® 25 TOTAL-D assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The DiaSorin LIAISON® 25 TOTAL-D is intended to be used on the LIAISON® XL Analyzer.
The DiaSorin LIAISON® 25 TOTAL-D Control Set is intended for use as assayed quality control samples to monitor the accuracy and precision of the DiaSorin LIAISON® 25 TOTAL-D assay.
The DiaSorin LIAISON® 25 TOTAL-D Calibration Verifiers are assayed quality control materials intended for use in the quantitative verification of calibration and reportable range of the LIAISON® 25 TOTAL-D assay when performed on the LIAISON® XL Analyzer.
The LIAISON® 25 TOTAL-D consists of one Reagent Intergral with calibrators, which consists of: Magnetic Particles (2.4 mL) coated with goat antibody against 25 OH Vitamin D, protein, phosphate buffer, < 0.1% sodium azide. Assay Buffer (28.0 mL) with 7.5% ethanol, surfactants and 0.2% ProClin® as a preservative. Conjugate (4.5 mL) 25 OH Vitamin D conjugated to an isoluminol derivative. in phosphate buffer with10% ethanol, EDTA and 0.1% benzoic acid as a preservative. Calibrator 1 (1.2 mL) Human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D. The calibrator concentrations (ng/mL) are referenced to standard preparations containing highly purified 25 OH Vitamin D. Calibrator 2 (1.2 mL) Human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D. The calibrator concentrations (ng/mL) are referenced to standard preparations containing highly purified 25 OH Vitamin D.
The LIAISON® 25 TOTAL-D control consists of 2 levels of human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D.
The LIAISON® 25 TOTAL-D calibration verifier consists of 4 levels of human serum, BSA. <0.1% sodium azide and 25 OH Vitamin D.
The DiaSorin LIAISON® 25 TOTAL-D assay is a chemiluminescent immunoassay (CLIA) for the quantitative determination of 25-hydroxyvitamin D and other hydroxylated vitamin D metabolites in human serum, EDTA, and Lithium heparin plasma. It is intended to be an aid in the assessment of vitamin D sufficiency in adults.
Here's an analysis of the acceptance criteria and the studies performed:
1. Table of Acceptance Criteria and Reported Device Performance
The acceptance criteria are not explicitly stated as pass/fail thresholds for each metric in the provided text. However, the study results are presented with implied acceptance based on the comparison to the predicate device and established clinical laboratory guidelines (e.g., CLSI documents).
| Performance Metric | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Method Comparison | Slope between 0.90-1.10 and high correlation to predicate. | N=391Slope: 0.99 (95% CI: 0.97-1.01)Intercept: -0.22 (95% CI: -0.51-0.08)Correlation Coefficient: 0.990 |
| Matrix Comparison (SST Serum vs. Serum) | Slope between 0.90-1.10 compared to serum. | Slope: 0.9942 (95% CI: 0.9785 to 1.0113)Y-intercept: 0.4098 (95% CI: -0.1694 to 1.1652) |
| Matrix Comparison (EDTA Plasma vs. Serum) | Slope between 0.90-1.10 compared to serum. | Slope: 1.0092 (95% CI: 0.9916 to 1.0253)Y-intercept: -0.5785 (95% CI: -1.1028 to -0.0976) |
| Matrix Comparison (Lithium Heparin Plasma vs. Serum) | Slope between 0.90-1.10 compared to serum. | Slope: 1.0007 (95% CI: 0.9869 to 1.0180)Y-intercept: -0.0211 (95% CI: -0.5554 to 0.4683) |
| Linearity/Measuring Range | Linear regression fit with statistically insignificant second and third order terms in serum, and a measuring range of 4-150 ng/mL. | Serum: y = 1.025x - 0.6094SST: y = 1.0064x - 0.3887EDTA: y = 1.0092x - 0.4958Li-Heparin: y = 1.0421x - 0.8295. Polynomial regression for serum showed statistically insignificant higher-order terms. Supports 4-150 ng/mL range. |
| LoB (Limit of Blank) | Not explicitly stated, but within expected assay limits. | < 2.03 ng/mL |
| LoD (Limit of Detection) | Not explicitly stated, but within expected assay limits. | 2.91 ng/mL |
| LoQ (Limit of Quantitation) | Not explicitly stated, but within expected assay limits. | 4.0 ng/mL |
| Recovery | Mean recovery close to 100%. | Mean Recovery: 96% |
| Interfering Substances | ≤ 10% bias relative to control for specified substances at given concentrations. | Met criteria for Hemoglobin (200 mg/dL), Triglycerides (638 mg/dL), conjugated/unconjugated Bilirubin (40 mg/dL), Cholesterol (350 mg/dL), Uric acid (20 mg/dL), Total Protein (12 g/dL), Biotin (0.1 mg/dL), Ascorbic Acid (6 mg/dL), Metoprolol (1.2 mg/dL), Propanolol hydrochloride (0.23 mg/dL), Furosemide (6 mg/dL), Hydrochlorothiazide (0.6 mg/dL), Paricalcital (0.012 ug/mL), Doxercalciferol (0.012 ug/mL). |
| Cross-reactivity | Acceptable levels of cross-reactivity with related steroids. | 25 OH Vitamin D3: 100%25 OH Vitamin D2: 93%Vitamin D3: 3.6%Vitamin D2: 1.9%1,25 (OH)2 Vitamin D3: 17.1%1,25 (OH)2 Vitamin D2: 27.1%3-epi-25 OH Vitamin D3: 1.9% |
| Reproducibility/Precision | Not explicitly stated, but low %CV within runs and overall. | Total %CV for controls: 11.6% (low), 6.2% (high)Total %CV for samples: 5.0% - 13.6% |
| Stability | Demonstrates stability for stated periods and conditions. | Controls & Cal Verifiers: Stable until expiration date when stored as instructed; 4 weeks opened at 2-8°C. Cal curve: 7 days. Open reagent vials: 4 weeks on-board/2-8°C. |
2. Sample Size Used for the Test Set and Data Provenance
- Method Comparison: 403 serum samples (391 included in analysis after excluding 12 below measuring range).
- Data Provenance: De-identified residual serum samples obtained from a sample procurement organization (U.S. and/or European patients). Retrospective.
- Matrix Comparison: 64 matched patient sets (serum, lithium heparin plasma, and EDTA plasma samples).
- Data Provenance: Not explicitly stated, but implied to be patient samples. Prospective.
- Reference Range Study: 395 samples.
- Data Provenance: Adults (21-90 years) recruited from four sites in the contiguous US (northern, southern, central US). Prospective.
- Reproducibility/Precision: 6 coded frozen serum samples (panel) and 2 kit controls.
- Data Provenance: Internal samples/controls.
- Dilution Linearity: Separate serum, SST (serum separator tubes), EDTA plasma, and lithium heparin plasma pools (original number of samples to create pools not specified).
- Data Provenance: Internal pools created for the study.
- LoB/LoD/LoQ:
- LoB: 6 blank sample aliquots.
- LoD: 4 samples.
- LoQ: 8 samples.
- Data Provenance: Internal samples.
- Recovery: 5 high concentration serum samples and 5 low concentration serum samples.
- Data Provenance: Internal samples.
- Interfering Substances: Vitamin D samples (30 ng/mL and 60 ng/mL) spiked with interfering substances.
- Data Provenance: Internal samples.
- Cross-reactivity: Three serum pools (15, 50, 75 ng/mL 25 OH D) spiked with potential cross-reactants.
- Data Provenance: Internal serum pools.
3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications of Those Experts
There is no mention of experts being used to establish ground truth in the traditional sense (e.g., radiologists interpreting images). For this type of in-vitro diagnostic device, the "ground truth" or reference values are established by other analytical methods or reference materials:
- Method Comparison: The predicate device, LIAISON® 25 OH Vitamin D TOTAL assay (K112725), served as the comparative "reference."
- Traceability and Value Assignment: Calibrators, controls, and calibration verifiers are traceable to UV spectrophotometric analysis of an in-house standard preparation using a commercially available 25-Hydroxyvitamin D. Master calibrators are prepared from a stock solution whose concentration is determined spectrophotometrically.
- Reference Range: The "ground truth" is the distribution of 25 OH Vitamin D levels in a statistically defined "apparently healthy" population, rather than expert consensus on individual cases.
4. Adjudication Method for the Test Set
Not applicable. This device is an in-vitro diagnostic (IVD) assay predicting a biochemical concentration, not a diagnostic imaging device requiring expert adjudication.
5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study Was Done
No. This is an in-vitro diagnostic device, not an imaging device that would typically involve human readers. Therefore, an MRMC study and analysis of AI vs. human reader improvement is not applicable.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done
Yes, the performance studies described (Method Comparison, Matrix Comparison, Linearity, LoB/LoD/LoQ, Recovery, Interfering Substances, Cross-reactivity, Reproducibility/Precision) evaluate the device's analytical performance in a standalone manner, without human interpretation of results beyond standard laboratory practices. The "algorithm" here is the chemiluminescent immunoassay technology performed on the LIAISON® XL Analyzer.
7. The Type of Ground Truth Used
The ground truth or reference values used for validation include:
- Comparison to a Predicate Device: For method comparison studies, the results from the previously cleared LIAISON® 25 OH Vitamin D TOTAL assay (K112725) served as the reference.
- Reference Standards/Spectrophotometric Analysis: For traceability, calibration, and control value assignment, established spectrophotometric methods using highly purified 25 OH Vitamin D standards were used.
- Defined Blanks and Spiked Samples: For LoB/LoD/LoQ, Recovery, Interfering Substances, and Cross-reactivity studies, samples with known (or zero) concentrations of the analyte or interfering substances were used.
- Population-Based Data: For reference range, the "ground truth" is derived from statistical analysis of a representative "apparently healthy" subject population.
8. The Sample Size for the Training Set
No explicit "training set" is mentioned in the context of machine learning. This device is a biochemical assay. The "training" for such devices typically involves:
- Method Development & Optimization: Internal studies and experiments conducted during the development phase to optimize reagent concentrations, incubation times, and other assay parameters.
- Calibration: The device uses a two-point calibrator to generate a stored master curve, which is essentially the "model" or "prediction function" for the assay. This calibration is derived from master calibrators whose values are assigned using reference materials.
9. How the Ground Truth for the Training Set Was Established
Not applicable in the machine learning sense. For the calibration and internal optimization processes:
- The ground truth for calibrators is established through UV spectrophotometric analysis of an in-house standard preparation using commercially available 25-Hydroxyvitamin D.
- Master calibrators are then used to assign values to the kit calibrators, controls, and calibration verifiers by running them on multiple LIAISON XL analyzers with different reagent lots and taking mean results.
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510(k) SUMMARY
SUBMITTED BY:
Mari Meyer Director, Regulatory Affairs DiaSorin Inc. 1951 Northwestern Avenue P.O. Box 285 Stillwater, MN 55082-0285 Phone (651) 439-9710 Fax (651) 351-5669 Email: mari.meyer@diasorin.com
SEP 0 5 2013
DATE PREPARED:
PURPOSE OF SUBMISSION:
NAME OF DEVICE:
Trade Name:
LIAISON® 25 TOTAL-D LIAISON® 25 TOTAL-D Control Set LIAISON® 25 TOTAL-D Calibration Verifiers
Common Names/Descriptions:
Classification Names:
Vitamin D
August 28, 2013
New Device
Vitamin D Test System: Class II 21 CFR 862.1825; Clinical Chemistry (75) Quality Control Material: Class I, reserved 21 CFR 862.1660; Clinical Chemistry (75)
Product Code:
PREDICATE DEVICE:
MRG, JJX
LIAISON® 25 OH Vitamin D TOTAL K112725 LIAISON® 25 OH Vitamin D Control K112725 LIAISON® 25 OH Vitamin D Calibration Verifier K090104
DEVICE DESCRIPTION:
The LIAISON® 25 TOTAL-D consists of one Reagent Intergral with calibrators, which consists of:
| Magnetic Particles | Magnetic particles coated with goat antibody against 25 |
|---|---|
| (2.4 mL) | OH Vitamin D, protein, phosphate buffer, < 0.1% sodium |
| azide. | |
| Assay Buffer | Buffer with 7.5% ethanol, surfactants and 0.2% ProClin® |
| (28.0 mL) | as a preservative. |
| Conjugate | 25 OH Vitamin D conjugated to an isoluminol derivative. |
| (4.5 mL) | in phosphate buffer with10% ethanol, EDTA and 0.1% |
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| benzoic acid as a preservative. | |
|---|---|
| Calibrator 1(1.2 mL) | Human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D. The calibrator concentrations (ng/mL) are referenced to standard preparations containing highly purified 25 OH Vitamin D. |
| Calibrator 2(1.2 mL) | Human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D. The calibrator concentrations (ng/mL) are referenced to standard preparations containing highly purified 25 OH Vitamin D. |
ProClin® 300 is a registered trademark of Rohm and Haas Co.
The LIAISON® 25 TOTAL-D control consists of 2 levels of human serum, BSA, <0.1% sodium azide and 25 OH Vitamin D.
The LIAISON® 25 TOTAL-D calibration verifier consists of 4 levels of human serum, BSA. <0.1% sodium azide and 25 OH Vitamin D.
INTENDED USE:
The LIAISON® 25 TOTAL-D assay is a chemiluminescent immunoassay (CLIA) intended for the quantitative determination of 25-hydroxyvitamin D and other hydroxylated vitamin D metabolites in human serum, EDTA and Lithium heparin plasma. The LIAISON® 25 TOTAL-D assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The DiaSorin LIAISON® 25 TOTAL-D is intended to be used on the LIAISON® XL Analyzer.
The DiaSorin LIAISON® 25 TOTAL-D Control Set is intended for use as assaved quality control samples to monitor the accuracy and precision of the DiaSorin LIAISON® 25 TOTAL-D assay.
The DiaSorin LIAISON® 25 TOTAL-D Calibration Verifiers are assayed quality control materials intended for use in the quantitative verification of calibration and reportable range of the LIAISON® 25 TOTAL-D assay when performed on the LIAISON® XL Analyzer.
TEST PRINCIPLE:
The DiaSorin LIAISON® 25 TOTAL-D assay is a direct competitive chemiluminescence immunoassay (CLA) for quantitative determination of 25 OH vitamin D in human serum or plasma. During the first incubation, 25 OH vitamin D is dissociated from its binding protein and binds to the specific antibody on the solid phases. After 10 minutes the tracer, (vitamin D linked to an isoluminol derivative) is added. After a second 10 minute incubation, the unbound material is removed with a wash cycle. Subsequently, the starter reagents are added to initiate a flash chemiluminescent reaction. The light signal is measured by a photomultiplier as relative light units (RLU) and is inversely proportional to the concentration of 25 OH vitamin D present in calibrators, controls, or samples.
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COMPARISON TO PREDICATE DEVICE:
The DiaSorin LIAISON 25 TOTAL-D assay is substantially equivalent in principle and performance to the LIAISON® 25 OH Vitamin D TOTAL assay (K112725) which was FDA cleared January 20, 2012.
| Table 1: Summary of Device Similarities - LIAISON® 25 TOTAL-D | |||
|---|---|---|---|
| Characteristic | New DeviceLIAISON® 25 TOTAL-D | Predicate DeviceLIAISON® 25 OH Vitamin DTOTAL assay (K112725) | |
| Intended Use | In-vitro assay for thequantitative determinationof 25-hydroxyvitamin D andother hydroxylated vitaminD metabolites. | In-vitro assay for thequantitative determination of25-hydroxyvitamin D andother hydroxylated vitamin Dmetabolites. | |
| Indications for Use | To be used as an aid in theassessment of vitamin Dsufficiency in adults. | Same | |
| Measured Analyte | 25-hydroxyvitamin D | Same | |
| Assay Type | ChemiluminescentImmunoassay | Same | |
| Test principle | CompetitivechemiluminescentImmunoassay | Same | |
| Solid Support | Paramagnetic particlescoated with goat antibodyagainst 25 OH Vitamin D | Same | |
| Reagent IntegralStorage | On-board or inrefrigerator@ 2-8°C | Same | |
| SampleHandling/Processing | Automated | Same | |
| Unit of Measure | ng/mL | Same | |
| Calibration | Two-point calibratorverification of stored mastercurve.Included with the kit. | Same | |
| Measuring range | 4 - 150 ng/mL | Same | |
| Conjugate Antibody | 25 OH Vitamin Dconjugated to an isoluminolderivative | Same | |
| Controls | 2 Levels | Same | |
| Open storage2-8°C | 4 weeks | Same | |
| Table 2 : Summary of Device Differences - LIAISON® 25 TOTAL-D | |||
| Characteristic | New DeviceLIAISON® 25 TOTAL-D | Predicate DeviceLIAISON® 25 OH Vitamin DTOTAL assay (K112725) | |
| Sample Matrix | Human serum, EDTA andLi heparin plasma | Human serum | |
| Sample size | 15 uL | 25 uL | |
| Calibration Matrix | Human serum with BSAand, sodium azide | Human serum with buffersalts and sodium azide. |
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The DiaSorin LIAISON 25 TOTAL-D Control is substantially equivalent in principle and performance to the LIAISON® 25 OH Vitamin D TOTAL Control (K112725) which was FDA cleared January 20, 2012.
| Table 3: Summary of Device Similairites and Difference LIAISON® 25 TOTAL-D Control Set | |||
|---|---|---|---|
| Characteristic | LIAISON® 25 TOTAL-DControl | LIAISON® 25 OH Vitamin DTOTAL Control (K112725) | |
| Intended Use | Intended for use asassayed quality controlsamples to monitor theaccuracy and precision ofthe assay. | Same | |
| Storage | Store at 2-8°C until readyto use | Same | |
| Matrix | Human serum with BSAand, sodium azide | Human serum with buffersalts and sodium azide. | |
| Levels | 2 levels: low and high | Same |
The DiaSorin LIAISON® 25 TOTAL-D Calibration Verifiers are substantially equivalent in principle and performance to the LIAISON® 25 OH Vitamin D TOTAL Calibration Verifiers (K090104) which were FDA cleared September 9, 2009.
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Table 4: Summary of Device Similarities and Differences LIAISON® 25 TOTAL-D Calibration Verifiers
| Characteristic | LIAISON® 25 TOTAL-DCalibration Verifiers | LIAISON® 25 OH Vitamin DTOTAL CalibrationVerifiers (K090104) |
|---|---|---|
| Intended Use | Assayed quality controlmaterials for in vitrodiagnostic use in thequantification verification ofcalibration and reportablerange of the DiaSorinLIAISON® 25 OH Vitamin DTOTAL Assay. | Same |
| Product Storage | 2 to 8°C | Same |
| Matrix | Human serum with BSAand, sodium azide | Human serum with buffersalts and sodium azide. |
| Levels | Four | Same |
| Volume | 5.0 mLs | Same |
PERFORMANCE DATA:
Method Comparison:
A method comparison study was performed on 403 serum samples following CLSI EP9-A2. The samples were tested in singlicate by both the LIAISON® 25 TOTAL-D assay and the predicate device. Testing for the LIAISON® 25 TOTAL-D assay and the predicate assay was performed at DiaSorin according to the respective Instructions for Use. All specimens were stored at -20°C or below until tested.
Samples tested in the method comparison included de-identified residual serum samples spanning the assay range which were obtained from a sample procurement organization. The samples are excess samples drawn from non-selected U.S. and/or European patients with sufficient volume to perform all testing required. Twenty-three (23) of the 403 samples were spiked or diluted samples in order to achieve sample values that reach the upper and/or lower limits of the assay range.
Three hundred ninety-one (391) of the 403 samples tested were analyzed. Twelve (12) samples read below the measuring range of the LIAISON® 25 TOTAL-D assay (<4.0 ng/mL) and therefore were not included in the analysis. Individual 25 OH Vitamin D results were plotted. Passing and Bablok regression analyses were performed for all samples across the measuring range of the assays.
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Results
The range of the results observed with the LIAISON® 25 TOTAL-D assay ranged from 4.18 - 135 ng/mL. Passing & Bablok regression analysis was applied to these samples and is summarized in the following table.
| N | Slope(95%CI) | Intercept(95% CI) | CorrelationCoefficient |
|---|---|---|---|
| 391 | 0.99(0.97-1.01) | -0.22(-0.51-0.08) | 0.990 |
Table 5: Method Comparison Passing & Bablok Regression Data
Matrix comparison:
Serum, SST Serum, Lithium Heparin, EDTA Plasma
Sixty four (64) matched patient sets of serum. lithium heparin plasma and EDTA plasma samples spanning the measuring range of the assay (4.0-139.3 ng/mL) were tested to determine if serum, SST serum, lithium heparin plasma and EDTA plasma samples provide equivalent results on the LIAISON® 25 TOTAL-D assay.
Results
A Passing-Bablok method comparison was used for analysis comparing each sample fype separately to the corresponding serum samples, with the specification of a 0.90 to 1.10 slope to serum considered acceptable.
| Bias | 95% CI | ||
|---|---|---|---|
| y-intercept | 0.4098 | -0.1694 | to 1.1652 |
| Slope | 0.9942 | 0.9785 | to 1.0113 |
SST Serum Compared to Serum
EDTA Plasma Compared to Serum
| Bias | 95% CI | ||
|---|---|---|---|
| y-intercept | -0.5785 | -1.1028 | to -0.0976 |
| Slope | 1.0092 | 0.9916 | to 1.0253 |
Lithium Heparin Plasma Compared to Serum
| Bias | 95% Cl | |||
|---|---|---|---|---|
| v-intercept | -0.0211 | -0.5554 | to 0.4683 | |
| Slope | 1.0007 | 0.9869 | to 1.0180 |
Conclusion
The study shows equivalent testing results from SST serum. EDTA plasma or lithium heparin plasma to serum.
Reference Range/Expected Values:
A reference range study was performed in accordance with CLSI Guideline C28-A2. Participant enrollment took place at four sites in the contiguous US, representing a wide distribution of sun strength: northern, southern, and central US. The collection was conducted on adults (ages 21 - 90 years) in summer and winter, including light and
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· dark skin individuals, and males and females. Three hundred and ninety five (395) samples met the inclusion/exclusion criteria and were tested with the LIAISON® 25 TOTAL-D Assav.
The determination of apparently healthy for this study was based on the following definition:
- · Normal serum levels of:
- o Total calcium
- o Intact parathyroid hormone (PTH)
- o Thyroid stimulating hormone (TSH)
- No personal history of kidney, gastrointestinal or liver disease .
- No personal history of parathyroid, thyroid, or chronic disease (defined by . severity of condition which is linked to treatment and frequency as stated on subject's CRF)
- No personal history of seizures .
- No bariatric surgery .
- No family history of parathyroid or calcium regulatory disease .
Other criteria were: at least 50% of subjects NOT taking ANY Vitamin D supplementation, and of the less than 50% of subject taking Vitamin D supplementation, level of supplementation must be <2000 IU/dav. Subjects were not currently taking any medications known to affect absorption (drugs that inhibit cholesterol absorption) or increase catabolism, such as anticonvulsants, glucocorticoids, HAART (AIDS treatment) and anti-reiection medications
-
- Current use of dietary or alternative therapies containing high concentrations of Vitamin D (e.g. adults ≥ 2.000 IU / day)
-
- More than 50% of subjects are taking Vitamin D supplementation
-
- Family history of parathyroid or calcium requlatory disease
-
- Personal history of the following diseases: kidney, gastrointestinal, liver, thyroid, or parathvroid
-
- Personal history of seizures
-
- Bariatric surgerv
-
- Pregnancy or lactation
The Instructions for Use contain the following table and statement: Reference Range
| Population(395) | Median25 OH Vitamin D | Observed Reference RangesCentral 95% Interval |
|---|---|---|
| United States | 22.9 ng/mL | 8.8 ng/mL – 54.2 ng/mL |
Consider these limits as guidelines only. It is important for each laboratory to establish its own reference range, representative of its typical population.
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Reproducibility/Precision:
A twenty day reproducibility/precision study was performed at DiaSorin Inc consisting of a coded panel comprised of 6 frozen serum samples. The coded panel contained samples for each level of low, medium and high 25 OH Vitamin D medical decision concentrations. The LIAISON® 25 TOTAL-D controls (2 levels) were also tested in the study. The CLSI document EP5-A2 was followed in the preparation of the testing protocol.
The precision panel samples and kit controls were tested on the LIAISON® 25 TOTAL-D assay on 1 Reagent Integral lot at one site in two replicates per run, 2 runs per day for 20 operating days on 1 analyzer for a total of 80 replicate results per sample.
The mean, standard deviation, and coefficient of variation (%CV) of the results were computed for each of the tested specimens.
Results
The 20 day results obtained at DiaSorin Inc. are summarized in Table 5 as sample overall mean 25 OH Vitamin D concentration in ng/mL, computed SDs and %CVs for within run and total within lot.
| SampleID | N | meanconc(ng/mL) | Within run | Total | ||
|---|---|---|---|---|---|---|
| KC 1 | 80 | 15.4 | 0.68 | 4.4% | 1.78 | 11.6% |
| KC 2 | 80 | 52.8 | 1.57 | 3.0% | 3.29 | 6.2% |
| Sample 1 | 80 | 11.2 | 0.43 | 3.8% | 1.52 | 13.6% |
| Sample 2 | 80 | 17.7 | 0.56 | 3.1% | 1.82 | 10.3% |
| Sample 3 | 80 | 28.2 | 0.80 | 2.9% | 2.36 | 8.4% |
| Sample 4 | 80 | 34.3 | 1.03 | 3.0% | 2.96 | 8.6% |
| Sample 5 | 80 | 65.1 | 1.35 | 2.1% | 3.97 | 6.1% |
| Sample 6 | 80 | 113.5 | 2.27 | 2.0% | 5.70 | 5.0% |
Table 7: DiaSorin Inc. - 20 dav Reproducibility/Precision
Dilution Linearity:
A lineanty study was performed using samples slightly above the upper measuring limit of the LIAISON® 25 TOTAL-D Assay, following CLSI EP6-A. Separate serum, SST (serum separator tubes), EDTA plasma and lithium heparin plasma pools that met this criterion were diluted with the assay's Specimen Diluent to yield multiple dilutions with Vitamin D levels spanning the assay's full measuring range of 4 - 150 ng/ml. Each dilution was analyzed in replicates of 4 with one lot of the modified Vitamin D Assay on one LIAISON® XL Analyzer.
Expected concentrations for the dilutions were determined by (Vitamin D value of high pool) x (dilution factor).
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Results
The results for each sample were analyzed by linear regression (slope and, intercept) of observed 25 OH D concentrations versus expected 25 OH D concentrations in ng/mL.
The resulting equation for each sample type is:
Serum: y (observed) = 1.025x (expected) - 0.6094 SST serum: y (observed) = 1.0064x (expected) - 0.3887 EDTA plasma: y (observed) = 1.0092x (expected) - 0.4958 Lithium heparin plasma: y (observed) = 1.0421x (expected) - 0.8295
A polynomial regression fit of the serum data yielded statistically insignificant second and third order terms. These results support the claimed measuring range of 4 to 150 ng/mL.
LoB/LoD/LoQ:
The study was performed based upon CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline June · 2012- Second Edition.
For LoB, 6 blank sample aliquots were tested in the LIAISON® 25 TOTAL-D assay on two analyzers, with two reagent lots and two technicians over six runs over three days yielding 60 concentration results.
For LoD, four (4) samples in the range of the mean LoB to 4 times the mean LoB were tested with the LIAISON® TOTAL-D on one LIAISON® XL Analyzer with two reagent . lots and two technicians over six runs and three days (2replicates/sample/run) vielding a minimum of 96 concentration results.
For LoQ, eight (8) samples (4 of which were used in the LoD determination) in the range of approximately 1.5 ng/mL to approximately 5.3 ng/mL were tested in the LIAISON® 25 TOTAL-D assay on two LIAISON® XL Analyzer with two reagent lots and two technicians over six runs and three days (2 replicates/sample/run), yielding 192 concentration results to define LoQ. To verify the LoQ, the same 8 samples were run on 1 LIAISON® XL Analyzer with a third reagent lot and two technicians over four runs (3 replicates/sample/run) yielding 96 concentration results to verify LoQ.
Results:
The limits are reported in the following table:
| LoB | LoD | LoQ |
|---|---|---|
| < 2.03 ng/mL | 2.91 ng/mL | 4.0 ng/mL |
Recovery:
Testing was performed to determine trueness or percent recovery on the LIAISON® 25 TOTAL-D. Five (5) high concentration serum samples (spiked or endogenous) and five (5) low concentration serum samples were analyzed on the LIAISON® 25 TOTAL-D assay.
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Recovery samples were then prepared by mixing defined ratios of the high and low samples and analyzing these in the LIAISON® 25 TOTAL-D assay. The obtained results were determined through calculation of % recovery for each dilution point:
% Recovery = (Observed conc. / Expected conc.)* 100
Expected conc. = conc. of High sample x % ratio used + Low sample x % ratio used
Each neat serum along with each blend was analyzed. The observed values were then compared to the expected values, based on the neat serum concentrations to determine the % recovery. The mean recovery is 96%.
| Serum Samples | DefinedConcentration | Expectedng/mL | Observedng/mL | %Recovery |
|---|---|---|---|---|
| Serum High Sample 1 (HS1) | 105.0 | |||
| 2 HS1 : 1LS1 | 73.5 | 72.3 | 98% | |
| 1 HS1 : 1LS1 | 57.2 | 54.5 | 95% | |
| 1 HS1 : 2LS1 | 41.0 | 38.3 | 93% | |
| Serum Low Sample 1 (LS1) | 9.5 | |||
| Serum High Sample 2 (HS2) | 95.5 | |||
| 2 HS2 : 1LS2 | 67.5 | 63.4 | 94% | |
| 1 HS2 : 1LS2 | 53.1 | 49.0 | 92% | |
| 1 HS2 : 2LS2 | 38.6 | 35.9 | 93% | |
| Serum Low Sample 2 (LS2) | 10.6 | |||
| Serum High Sample 3 (HS3) | 101.0 | |||
| 2 HS3 : 1LS3 | 70.8 | 68.7 | 97% | |
| 1 HS3 : 1LS3 | 55.3 | 52.4 | 95% | |
| 1 HS3 : 2LS3 | 39.7 | 37.3 | 94% | |
| Serum Low Sample 3 (LS3) | 9.5 | |||
| Serum High Sample 4 (HS4) | 58.5 | |||
| 2 HS4 : 1LS4 | 44.4 | 43.7 | 98% | |
| 1 HS4 : 1LS4 | 37.1 | 36.4 | 98% | |
| 1 HS4 : 2LS4 | 29.8 | 29.3 | 98% | |
| Serum Low Sample 4 (LS4) | 15.7 | |||
| Serum High Sample 5 (HS5) | 56.9 | |||
| 2 HS5 : 1LS5 | 42.2 | 42.2 | 100% | |
| 1 HS5 : 1LS5 | 34.7 | 33.9 | 98% | |
| 1 HS5 : 2LS5 | 27.1 | 26.9 | 99% | |
| Serum Low Sample 5 (LS5) | 12.4 | |||
| Mean Recovery | 96% |
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Interfering Substances:
Vitamin D samples containing concentrations of 30 ng/mL and 60 ng/mL were spiked with hemoglobin, trialycerides, cholesterol, and bilirubin, uric acid, albumin.lgG, biotin, ascorbic acid, metoprolol, propanolol hydrochloride, furosemide, hydrochlorothiazide, paricalcital and doxercalciferol. Replicate samples (n=6 for each test and control sample) were tested and results were compared to control samples without the potentially interfering substances added. The highest concentrations at which no interference, or minimal interference (defined by the sponsor as ≤ 10% bias relative to the control) are shown below. The testing was based on CLSI EP7-A2.
| Specimens That Are | Demonstrate < 10% change in Results Up To |
|---|---|
| Hemolyzed | 200 mg/dL of hemoglobin |
| Lipemic | 638 mg/dL of triglycerides |
| Icteric | 40 mg/dL of conjugated bilirubin |
| Icteric | 40 mg/dL of unconjugated bilirubin |
Specimens That Contain
Cholesterol Uric acid Total Protein Biotin Ascorbic Acid Metoprolol Propanolol hydrochloride Furosemide Hydrochlorothiazide Paricalcital Doxercalciferol
Demonstrate < 10% change in Results Up To 350 mg/dL 20 mg/dL 12 g/dL 0.1 mg/dL 6 mg/dL 1.2 mg/dL 0.23 mg/dL 6 mg/dL 0.6 mg/dL 0.012 ua/mL
Cross-reactivity:
Cross-reactivity studies were performed as described in CLSI EP7-A2 using three serum pools at 25 OH D concentration levels of 15 ng/mL, 50 ng/mL, and 75 ng/mL. Data on the cross-reactivity of the antiserum used in this assay were obtained by spiking up to 100 ng/mL of the potential cross-reactant and assaying. The crossreactivity of each compound, normalized to 25 OH Vitamin D3, is listed below.
0.012 ug/mL
| Steroid | % Crossreactivity |
|---|---|
| 25 OH Vitamin D3 | 100% |
| 25 OH Vitamin D2 | 93% |
| Vitamin D3 | 3.6% |
| Vitamin D2 | 1.9% |
| 1,25 (OH)2 Vitamin D3 | 17.1% |
| 1,25 (OH)2 Vitamin D2 | 27.1% |
| 3-epi-25 OH Vitamin D3 | 1.9% |
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Stability:
Shelf life stability studies were performed with controls and calibration verifiers and demonstrated that they are stable until the expiration date shown on the product labeling when stored as instructed. Controls and calibration verifiers are stable until the expiration date printed on the label when stored as directed. Once opened, controls and calibration verifiers are stable for 4 weeks when properly stored at 2-8℃ between uses. Calibration curve stability and reagent open vial stability were performed and demonstrated that the calibration curve is stable for 7 days and open reagent vials are stable for 4 weeks when stored on board or at 2-8°C.
Traceability
The LIAISON® 25 TOTAL-D Calibrators. Controls and Calibration Verifiers are traceable to UV spectrophotometric analysis of an in-house standard preparation. The standard material used is a commercially available 25-Hydroxyvitamin D. The stock solution of 25 0H Vitamin Dis then used to make calibrators by diluting into human serum.
Value assignment:
Concentrations of calibrators controls and calibration verifiers are assigned through an internal procedure. Master calibrators are prepared from a stock solution made from reference material whose concentration is determined spectrophotometrically. The master calibrators are then used to assign values to the kit calibrators, controls and calibration verifiers using a minimum of 3 LIAISON XL analyzers with at least 2 reagent lots. Each lot of controls and calibration verifiers are tested over several runs and the mean results are used to determine the target values.
25 TOTAL-D Calibrators Level 1= 3.2 -4.8 ng/mL · Level 2= 70.0 – 80.0 ng/mL
25 TOTAL-D Controls have the following target ranges: Level 1= 12.0 - 18.0 ng/mL Level 2= 40.0 - 60.0 ng/mL
25 TOTAL-D Calibration Verifiers have the following target ranges: Level 1= 8.5 - 11.5 ng/mL Level 2= 27.0 - 33.0 ng/mL Level 3= 59.0 - 71.0 ng/mL Level 4= 108 - 122 ng/mL
CONCLUSION:
The material submitted in this premarket notification is complete and supports the basis for substantial equivalence to the LIAISON® 25 OH Vitamin D TOTAL assay (K112725).
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DEPARTMENT OF HEALTH & HUMAN SERVICES
Image /page/12/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features a stylized eagle with three wavy lines representing the agency's mission. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" is arranged in a circular pattern around the eagle.
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Control - WO66-G609 Silver Spring, MD 20993-0002
September 5, 2013
DiaSorin Inc. C/O Mari Mever P. O. Box 285 1951 Northwestern Avenuc STILLWATER MN 55082-0285
Re: K132492 Trade Name: LIAISON® 25 TOTAL-D LIAISON® 25 TOTAL-D Control Set LIAISON® 25 TOTAL-D Calibration Verifiers Regulation Number: 21 CFR $862.1825 Regulation Name: Vitamin D Test System Regulatory Class: Class II Product Codes: MRG, JJX Dated: July 29, 2013 Received: August 9, 2013
Dear Ms. Meyer:
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. Ilsting 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 Part 801); 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 regulation (21 CFR Part 801), please go to http://www.fda.gov/AboutFDA/CentersOfTices/CDRH/CDRHQ0ffices/ucm115809.html for the Center for Devices and Radiological Health's (CDRH's) Office of Compliance. 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/ReportalProblem/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/Resourcesfor You/Industry/default.htm.
Sincercly vours.
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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Indication for Use
510(k) Number (if known): K132492
LIAISON® 25 TOTAL-D, LIAISON® 25 TOTAL-D Control Set and Device Name: LIAISON® 25 TOTAL-D Calibration Verifiers
The LIAISON® 25 TOTAL-D assay is a chemiluminescent immunoassay (CLIA) intended for the quantitative determination of 25-hydroxyvitamin D and other hydroxylated vitamin D metabolites in human serum. EDTA and Lithium heparin plasma. The LIAISON® 25 TOTAL-D assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The DiaSorin LIAISON® 25 TOTAL-D is intended to be used on the LIAISON® XL Analyzer.
The DiaSorin LIAISON® 25 TOTAL-D Control Set is intended for use as assayed quality control samples to monitor the accuracy and precision of the DiaSorin LIAISON® 25 TOTAL-D assay.
The DiaSorin LIAISON® 25 TOTAL-D Calibration Verifiers are assayed quality control materials intended for use in the quantitative verification of calibration and reportable range of the LIAISON® 25 TOTAL-D assay when performed on the LIAISON® XL Analyzer.
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)
Yung.W.Chan -S
Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) kl32492
§ 862.1825 Vitamin D test system.
(a)
Identification. A vitamin D test system is a device intended for use in clinical laboratories for the quantitative determination of 25-hydroxyvitamin D (25-OH-D) and other hydroxylated metabolites of vitamin D in serum or plasma to be used in the assessment of vitamin D sufficiency.(b)
Classification. Class II (special controls). Vitamin D test systems must comply with the following special controls:(1) Labeling in conformance with 21 CFR 809.10 and
(2) Compliance with existing standards of the National Committee on Clinical Laboratory Standards.