ELECSYS CALCITONIN; IMMUNOASSAY, CALSET, CALCHECK 5; ELECSYS PRECICONTROL VARIA 3
Device Facts
| Record ID | K132828 |
|---|---|
| Device Name | ELECSYS CALCITONIN; IMMUNOASSAY, CALSET, CALCHECK 5; ELECSYS PRECICONTROL VARIA 3 |
| Applicant | Roche Diagnostics |
| Product Code | JKR · Clinical Chemistry |
| Decision Date | Dec 4, 2013 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.1140 |
| Device Class | Class 2 |
Indications for Use
Elecsys Calcitonin Reagent: • The Calcitonin immunoassay is intended for the in vitro quantitative determination of human calcitonin (thyrocalcitonin) in serum and plasma. The calcitonin determination is intended to be used as an aid in the diagnosis and treatment of diseases involving the thyroid and parathyroid glands, including carcinoma and hyperparathyroidism in conjunction with other clinical and laboratory findings. The electrochemiluminescence immunoassay “ECLIA” is intended for use on the indicated Elecsys and cobas e immunoassay analyzers. Elecsys Calcitonin CalSet: • Calcitonin CalSet is used for calibrating the quantitative Elecsys Calcitonin assay on the Elecsys and cobas e immunoassay analyzers. Elecsys Calcitonin CalCheck 5: • The Elecsys Calcitonin CalCheck 5 is an assayed control for use in calcitonin calibration verification and for use in the verification of the calcitonin assay range established by the Elecsys and cobas e immunoassay analyzers. Elecsys PreciControl Varia: • The Elecsys PreciControl Varia is used for quality control of the Elecsys immunoassays on Elecsys and cobas e immunoassay analyzers.
Device Story
The Elecsys Calcitonin Immunoassay is an in vitro diagnostic test for human calcitonin in serum and plasma. It uses a sandwich electrochemiluminescence immunoassay (ECLIA) principle; murine monoclonal antibodies directed against hCT are labeled with a ruthenium complex and biotin, respectively, with streptavidin-coated microparticles as the solid phase. The device is intended for use on Elecsys and cobas e immunoassay analyzers by laboratory professionals. Results are determined via a calibration curve generated by a 2-point calibration and a master curve provided with the reagent barcode. The output provides quantitative calcitonin levels, which clinicians use alongside other findings to diagnose and manage thyroid and parathyroid conditions. The system includes dedicated calibrators (CalSet), calibration verification controls (CalCheck 5), and multi-analyte quality controls (PreciControl Varia).
Clinical Evidence
No clinical diagnostic studies performed. Evidence consists of analytical bench testing: precision (total CV 2.6-6.5%), linearity (1.0-2000 pg/mL), LoB/LoD/LoQ (0.3/0.5/1.0 pg/mL), and method comparison against predicate (n=150, Deming regression y=0.998x-3.05, r=0.991). Interference and cross-reactivity studies confirmed assay robustness.
Technological Characteristics
Sandwich ECLIA; murine monoclonal antibodies; ruthenium complex/biotin labels; streptavidin-coated microparticles. Analyzers: cobas e 411. Calibrators/Controls: Lyophilized equine serum matrix. Connectivity: Barcode-driven master curve. Standards: CLSI EP5-A2 (precision), EP17-A/A2 (LoD/LoQ), EP6-A (linearity), EP09-A2-IR (method comparison).
Indications for Use
Indicated for in vitro quantitative determination of human calcitonin in serum and plasma to aid in diagnosis and treatment of thyroid and parathyroid gland diseases, including carcinoma and hyperparathyroidism, in conjunction with other clinical and laboratory findings.
Regulatory Classification
Identification
A calcitonin test system is a device intended to measure the thyroid hormone calcitonin (thyrocalcitonin) levels in plasma and serum. Calcitonin measurements are used in the diagnosis and treatment of diseases involving the thyroid and parathyroid glands, including carcinoma and hyperparathyroidism (excessive activity of the parathyroid gland).
Predicate Devices
- Siemens Immulite 1000 Calcitonin, Model LKCL (k023304)
- Elecsys Vitamin D CalSet (k113546)
- Elecsys T4 CalCheck 5 (k112528)
- Elecsys PreciControl Varia (k111506)
Submission Summary (Full Text)
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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE
A. 510(k) Number:
k132828
B. Purpose for Submission:
New Device
C. Measurand:
Calcitonin
D. Type of Test:
Quantitative, Electrochemiluminescence Immunoassay (ECLIA)
E. Applicant:
Roche Diagnostics
F. Proprietary and Established Names:
Elecsys Calcitonin Immunoassay
Elecsys Calcitonin CalSet
Elecsys Calcitonin CalCheck5
Elecsys PreciControl Varia
G. Regulatory Information:
1. Regulation section:
21 CFR 862.1140, Radioimmunoassay, Calcitonin
21 CFR 862.1150, Calibrator, Secondary
21 CFR 862.1660, Quality control material
2. Classification:
Class II
Class II
Class I, reserved
3. Product code:
JKR
JIT
JJY
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4. Panel:
Chemistry (75)
H. Intended Use:
1. Intended use(s):
See Indication(s) for use
2. Indication(s) for use:
Elecsys Calcitonin Immunoassay:
The Calcitonin Immunoassay is intended for the in vitro quantitative determination of human calcitonin (thyrocalcitonin) in serum and plasma. The calcitonin determination is intended to be used as an aid in the diagnosis and treatment of diseases involving the thyroid and parathyroid glands, including carcinoma and hyperparathyroidism in conjunction with other clinical and laboratory findings. The Electrochemiluminescence immunoassay "ECLIA" is intended for use on the indicated Elecsys and cobas e immunoassay analyzers.
Elecsys Calcitonin CalSet:
Calcitonin CalSet is used for calibrating the quantitative Elecsys Calcitonin assay on the Elecsys and cobas e immunoassay analyzers.
Elecsys Calcitonin CalCheck 5:
The Elecsys CalCheck 5 is an assayed control for use in calibration verification and for use in the verification of the calcitonin assay range established by the Elecsys and cobas e immunoassay analyzers.
Elecsys PreciControl Varia:
The Elecsys PreciControl Varia is used for quality control of the Elecsys immunoassays on Elecsys and cobas e immunoassay analyzers.
3. Special conditions for use statement(s):
For prescription use only.
4. Special instrument requirements:
cobas e 411 Immunoassay Analyzer
I. Device Description:
The Elecsys Calcitonin Reagent kit consists of 3 working solutions:
M- Streptavidin-coated microparticles (0.72 mg/mL) and preservative.
R1- Anti-hCT-Ab-biotin: Biotinylated monoclononal anti-hCT antibody (mouse), phosphate buffer and preservative.
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R2- Anti-hCT-Ab~Ru(bpy): monoclonal anti-hCT antibody (mouse) labeled with ruthenium complex; phosphate buffer, and preservative.
The Elecsys Calcitonin CalSet consists of 2 levels, hCT Cal 1 and hCT Cal 2. The Calcitonin calibrator set contains lyophilized calcitonin (synthetic) in two concentration ranges (approximately 2.0 pg/mL and 500 pg/mL) in an equine serum matrix.
The Elecsys Calcitonin CalCheck 5: Each set contains 5 lyophilized levels of human calcitonin (synthetic) in buffered equine serum. The approximate target range for each level is listed below:
Check 1 ≤1.00 pg/mL
Check 2 34.5-65.5 pg/mL
Check 3 345-655 pg/mL
Check 4 690-1310 pg/mL
Check 5 1380->2000 pg/mL
PreciControl Varia is a multi-composite lyophilized 3 level control set based on human source material. The PreciControl V1 and V2 level contains lyophilized calcitonin (synthetic) in two concentration ranges (approximately 10 pg/mL and 100 pg/mL). The PreciControl V0 does not contain any calcitonin.
All products derived from human blood are prepared exclusively from the blood of donors tested individually by FDA approved methods and shown to be free from HbsAg and antibodies to HCV and HIV.
J. Substantial Equivalence Information:
1. Predicate device name(s):
Immulite 1000 Calcitonin
Elecsys Vitamin D CalSet
Elecsys T4 CalCheck 5
Elecsys PreciControl Varia
2. Predicate 510(k) number(s):
k023304
k113546
k112528
k111506
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3. Comparison with predicate:
Assay Reagents
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Device Elecsys Calcitonin Immunoassay | Predicate Immulite 1000 Calcitonin (k023304) |
| Intended Use | Immunoassay for the in vitro quantitative determination of human calcitonin (thyrocalcitonin) in serum and plasma. | Same |
| Traceability | Standardized against the WHO 2^{nd} IRP 89/620 | Same |
| | Electrochemiluminescent Immunoassay | Chemiluminescent Immunometric assay |
| Reaction time | 18 minutes | 60 minutes |
| Sample type | Human serum and plasma treated with K2-EDTA, K3-EDTA, lithium heparin and lithium heparin plasma gel separation tubes. | Human serum and heparinized plasma |
| Sample volume | 50 μL | 75 μL |
| Calibration interval | 2 months when using the same reagent lot. 7 days when using the same reagent kit on the analyzer. | 4 weeks |
| Reagent Stability | Unopened: 2-8°C - Up to the stated expiration date Opened 2-8°C - 12 weeks On Analyzers – 4 weeks | Stable at 2-8°C until expiration date |
| Analytical Sensitivity | 1 pg/mL | 2 pg/mL |
CalSet (calibrator)
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Device Elecsys Calcitonin CalSet | Predicate Elecsys Vitamin D CalSet (k133546) |
| Intended Use | For calibrating the assay on the Elecsys and cobas e immunoassay analyzers. | Same |
| Levels | Two | Same |
| Format | Lyophilized | Same |
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| Similarities and Differences | | |
| --- | --- | --- |
| Item | Device Elecsys Calcitonin CalSet | Predicate Elecsys Vitamin D CalSet (k133546) |
| Matrix | Buffered (50 mmol/L HEPES) equine serum | Human serum |
| Stability | Unopened: up to the stated expiration date
After reconstituting:
At 2-8°C – 72 hours
At -20°C – 84 days (freeze only once)
At 20-25°C: up to 5 hours | Unopened: up to the stated expiration date
After reconstituting:
At 2-8C – 120 hours
At -20C – 90 days (freeze only once)
On Elecsys 2010/cobas e 411 at 20-25°C: up to 5 hours |
CalCheck (Calibration Verifiers)
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Device Elecsys Calcitonin CalCheck 5 | Predicate Elecsys T4 CalCheck5 (k112528) |
| Intended Use | It is an assayed control for use in calibration verification and for use in the verification of the assay range. | Same |
| Levels | Five | Same |
| Format | Lyophilized | Same |
| Matrix | Buffered (50 mmol/L HEPES) equine serum | Level 1: BSA
Level 2-5:Human serum |
| Stability | Unopened: store at 2-8°C up to the stated expiration date
After reconstituting:
At 20-25°C: up to 4 hours | Unopened: store at 2-8°C up to the stated expiration date
After reconstituting:
At 15-25°C : up to 5 hours |
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Controls (PreciControl Varia)
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Device Elecsys PreciControl Varia | Predicate Elecsys PreciControl Varia (k111506) |
| Intended Use | It is used for quality control of Elecsys immunoassays on the Elecsys and cobas e immunoassay analyzers | Same |
| Volume (reconstituted) | 3.0 mL | Same |
| Stability | Unopened: store at 2-8°C up to the stated expiration date
After reconstituting:
At -20°C: 31 days
At 2-8°C: 72 hours
At 20-25°C: on board the analyzer: up to 5 hours | Same |
| Calcitonin Concentration | Level 1: 10 pg/mL
Level 2: 100 pg/mL | No calcitonin concentration present |
## K. Standard/Guidance Document Referenced (if applicable):
CLSI EP5-A2 Evaluation of Precision of Clinical Chemistry Devices; Approved Guideline 2nd Edition
CLSI EP17-A Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline, 1st Edition
CLSI EP6-A Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline, 1st Edition
CLSI EP17-A2 Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline 2nd Edition
CLSI EP09-A2-IR Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline, 2nd Edition
## L. Test Principle:
The Roche Calcitonin Immunoassay is an Electrochemiluminescence Immunoassay (ECLIA) which is based on the sandwich principle. During the first incubation, 50 μL of sample, a biotinylated monoclonal hCT-specific antibody and a monoclonal hCT-specific antibody labeled with a ruthenium complex react to form a sandwich complex. During the second incubation, streptavidin-coated microparticles are added and form a complex which becomes bound to the solid phase via interaction of biotin and streptavidin. The reaction mixture is aspirated into the measuring cell where the
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microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell/ProCell M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier. Results are determined via a calibration curve which is instrument specifically generated by 2-point calibration and a master curve provided via the reagent barcode.
## M. Performance Characteristics (if/when applicable):
### 1. Analytical performance:
#### a. Precision/Reproducibility:
Precision studies were performed on one cobas e 411 Immunoassay analyzer with one reagent lot. The samples consisted of PreciControl Varia and human sera pools. Two replicates of each control and human sera were run per day for 21 days for a total of 84 replicates. Unaltered serum sample pools were used in addition to spiked and diluted pools in order to obtain samples to cover the measuring range. CLSI EP5-A2 guidelines were followed. The precision results are summarized in the table below:
| | | | Within-run | | Total precision | |
| --- | --- | --- | --- | --- | --- | --- |
| Sample | n | Mean (pg/mL) | SD (pg/mL) | CV (%) | SD (pg/mL) | CV (%) |
| Human serum 1 | 84 | 1.21 | 0.034 | 2.8 | 0.043 | 3.6 |
| Human serum 2 | 84 | 11.5 | 0.345 | 3.0 | 0.413 | 3.6 |
| Human serum 3 | 84 | 48.5 | 1.24 | 2.5 | 1.71 | 3.5 |
| Human serum 4 | 84 | 482 | 13.8 | 2.9 | 19.2 | 4.0 |
| Human serum 5 | 84 | 1910 | 42.6 | 2.2 | 65.0 | 3.4 |
| PC Varia 1 | 84 | 8.88 | 0.191 | 2.1 | 0.261 | 2.9 |
| PC Varia 2 | 84 | 97.7 | 1.44 | 1.5 | 2.51 | 2.6 |
Reproducibility of the Elecsys Calcitonin assay was evaluated over three sites using three different cobas e 411 Immunoassay analyzers based on CLSI EP5-A2 guidelines. Sources of variability included testing for 21 days, 2 runs per day, with 2 replicates for each panel member per run at three sites. One reagent lot was evaluated. Two controls (PCV = PreciControl Varia, Level 1 & 2) and five human serum pools (HSP) were tested. The human serum pools used were native as well as spiked and diluted.
The following precision study result summary is from one site and is representative of the results obtained from all 3 sites.
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| Sample | Mean (pg/mL) | Within-run | | Total precision | | n |
| --- | --- | --- | --- | --- | --- | --- |
| | | SD (pg/mL) | CV (%) | SD (pg/mL) | CV (%) | |
| HSP01 | 1.11 | 0.0377 | 3.4 | 0.0596 | 5.4 | 84 |
| HSP02 | 12.7 | 0.387 | 3.0 | 0.464 | 3.7 | 84 |
| HSP03 | 52.6 | 1.32 | 2.5 | 1.87 | 3.6 | 84 |
| HSP04 | 514 | 17.0 | 3.3 | 18.6 | 3.6 | 84 |
| HSP05 | 1992 | 50.6 | 2.5 | 66.7 | 3.3 | 84 |
| PCV L1 | 9.76 | 0.204 | 2.1 | 0.307 | 3.1 | 84 |
| PCV L2 | 106 | 1.82 | 1.7 | 2.74 | 2.6 | 84 |
Summary of Reproducibility study over 3 sites
| Sample | Repeatability | | Between-run | | Between-day | | Between-lab | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Mean pg/mL | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| HSP1 1.06 | 0.03 | 3.1 | 0.03 | 2.6 | 0.03 | 3.3 | 0.04 | 3.8 | 0.07 | 6.5 |
| HSP2 12.4 | 0.27 | 2.2 | 0.14 | 1.1 | 0.22 | 1.8 | 0.26 | 2.1 | 0.46 | 3.7 |
| HSP3 51.2 | 0.99 | 1.9 | 0.85 | 1.7 | 0.74 | 1.4 | 1.19 | 2.3 | 1.92 | 3.7 |
| HSP4 500 | 12.0 | 2.4 | 4.89 | 1.0 | 8.2 | 1.6 | 11.8 | 2.4 | 19.4 | 3.9 |
| HSP5 1924 | 36.8 | 1.9 | 31.2 | 1.6 | 24.7 | 1.3 | 58.7 | 3.1 | 79.9 | 4.2 |
| PCV L1 9.32 | 0.15 | 1.6 | 0.12 | 1.3 | 0.19 | 2.0 | 0.39 | 4.1 | 0.47 | 5.1 |
| PCV L2 101 | 1.48 | 1.5 | 1.38 | 1.4 | 1.6 | 1.6 | 4.21 | 4.2 | 4.94 | 4.9 |
b. Linearity/assay reportable range:
Linearity of the Elecsys Calcitonin assay was assessed on the cobas e 411 Immunoassay analyzer according to CLSI EP6-A. A high analyte serum sample pool (spiked) was diluted with a low analyte serum sample pool. Twenty-one concentrations throughout the measuring range were prepared. Samples were assayed in 3-fold determination within a single run. Samples tested range between 0.28 to 2186.5 pg/mL.
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The calcitonin linearity study resulted in a polynomial regression in which the 2nd order polynomial regression was considered significant and yielded a best fit square linearity equation of $y = 0.9707x + 0.0617$.
The linearity study supports the sponsor’s claimed measuring range: 1.0 – 2000 pg/mL.
**Hook Effect**: To evaluate the hook effect, two human serum samples were spiked with analyte concentration up to 1,200,000 pg/mL. The samples were assessed on the cobas e 411. It was determined that here is no high-dose hook effect at calcitonin concentrations up to 1,000,000 pg/mL.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
**Traceability**:
The Elecsys Calcitonin CalSet, Elecsys Calcitonin CalCheck 5, and the calcitonin component of the PreciControl Varia are traceable to the WHO 2nd IRP 89/620 Reference Standard.
**Elecsys Calcitonin CalSet Stability**: Real-time and accelerated stability studies were performed to determine the Elecsys Calcitonin CalSet stability. The study protocol and acceptance criteria were reviewed and found to be acceptable. The Elecsys Calcitonin CalSet stability studies support the following stability claims: Unopened: up to 12 months when stored at 2-8°C. After reconstituting: At 2-8°C, 72 hours; at -20°C, 84 days, at 20-25°C: up to 5 hours.
**Elecsys Calcitonin CalCheck5 Stability**: Real-time and accelerated stability studies were performed to determine Elecsys Calcitonin CalCheck 5 stability. The study protocol and acceptance criteria were reviewed and found to be acceptable. The Elecsys Calcitonin CalCheck 5 stability studies support the following stability claims: Unopened: 12 months stored at 2-8°C. Reconstituted: 20-25°C: up to 4 hours
**Elecsys PreciControl Varia Stability**: Real-time and accelerated stability studies were performed to determine Elecsys Calcitonin CalCheck 5 stability. The study protocol and acceptance criteria were reviewed and found to be acceptable. The Elecsys PreciControl Varia stability studies support the following stability claims: Unopened at 2 – 8 °C, up to 15 months; reconstituted/thawed serum at -20 °C, for 31 days (freeze only once); at 2 – 8 °C, for 72 hours; and at 20 – 25 °C on-board the analyzers, up to 5 hours.
**Value assignment for ElecsysCalcitonin CalSet, Elecsys Calcitonin CalCheck 5, and PreciControl Varia**:
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Value assignment is determined by performing multiple runs using multiple analyzers and the mean results are calculated based on the multiple replicate results. The target values for the ElecsysCalcitonin CalSet, Elecsys Calcitonin CalCheck 5, and PreciControl Varia are listed in the tables below:
Target Values for Elecsys Calcitonin CalSet
| Level | Calcitonin Target Value (pg/mL) | Calcitonin Target Range (pg/mL) |
| --- | --- | --- |
| Calibrator 1 | 2.0 | 1.5 – 2.5 |
| Calibrator 2 | 500 | 400 – 600 |
Target Values for the Elecsys Calcitonin CalCheck5
| Level | Calcitonin Target Value (pg/mL) | Calcitonin Target Range (pg/mL) |
| --- | --- | --- |
| Check 1 | <1 | - |
| Check 2 | 50 | 45-55 |
| Check 3 | 500 | 450-550 |
| Check 4 | 1000 | 900-1100 |
| Check 5 | 2000 | 1800-2200 |
Target Values for Elecsys PreciControl Varia
| Control Level | Calcitonin Target Value (pg/mL) | Target Range (pg/mL) |
| --- | --- | --- |
| Level 1 | 10.0 | 8.9-12.1 |
| Level 2 | 100 | 89-121 |
d. Detection limit
Limit of Blank (LoB) was determined using three reagent lots on two cobas e 411 analyzers, measuring five zero-level human serum samples during three days, two runs per day for a total of 60 replicates per sample per reagent lot. The LoB was determined as the $95^{\text{th}}$ percentile of measurement of blank samples.
Limit of Detection (LoD) was determined using three reagent lots on two cobas e 411 analyzers, measuring five low-level human serum samples during three days, two runs per day for a total of 60 replicates per sample per reagent lot. The LoD was determined as the lowest amount of analyte in a sample that can be detected with $95\%$ probability.
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Limit of Quantitation (LoQ) was determined using three reagent lots on two cobas e 411 analyzers, measuring nine low-level human serum samples during three days, two runs per day for a total of 60 replicates per sample per reagent lot. The LoQ was derived from a plot of the allowable error versus the expected concentration of Calcitonin at a total error of 20%.
The detection limits are summarized in the table below:
Elecsys Calcitonin Immunoassay
| Limit of Blank (LoB) | Limit of Detection (LoD) | Limit of Quantitation (LoQ) |
| --- | --- | --- |
| 0.3 pg/mL | 0.5 pg/mL | 1.0 pg/mL |
e. Analytical specificity:
Cross Reactivity: The specificity of the Elecsys Calcitonin assay was determined using two human serum sample pools spiked with potential cross-reactant compounds. The sample analyte concentrations were approximately 10 and 500 pg/mL calcitonin. The spiked and non-spiked samples were tested in duplicate on the cobas e 411 Immunoassay analyzer. The cross-reactivity results are summarized in the table below:
| Cross reactant | Max. concentration tested (ng/mL) | Highest cross-reactivity observed (%) |
| --- | --- | --- |
| Salmon Calcitonin | 200 | 0.017 |
| Porcine Calcitonin | 1000 | 0.007 |
| Chicken Calcitonin | 1000 | 0.005 |
| ACTH (1-39) human | 200 | 0.037 |
| C-Peptide | 80000 | 0.000 |
| Calcitonin Gene Related Peptide | 2000 | 0.002 |
| PTH (1-84) | 300 | 0.013 |
| TSH | 2000μIU/mL | 0.009 |
| Insulin | 67000 | 0.000 |
| Prolactin | 2000 | 0.001 |
| Gastrin I | 4000 | 0.001 |
| Elcatonin | 200000 | 0.000 |
| Katacalcin | 80000 | 0.000 |
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Endogenous Interference: The effect on quantitation of analyte in the presence of endogenous interfering substances using the Elecsys Calcitonin was determined on the cobas e 411 immunoassay analyzer for the following 8 interfering substances: Hemoglobin, Biotin, Lipemia, Bilirubin, Rheumatoid Factor, IgG, IgA, and IgM. For each interfering substance, three spiked human serum samples (one low, one medium, and one high concentrations of Calcitonin) were used to prepare dilution series of 11 dilutions that were tested in duplicate. The sponsor defined significant interference as greater than $\pm 10\%$ of the unspiked (reference) value. The endogenous interference effects on analyte quantitation are summarized in the table below:
| Substance | Highest Concentration at which no significant interference was observed. |
| --- | --- |
| Hemoglobin | 200 mg/dL |
| Biotin | 40 ng/dL |
| Intralipid (lipemia) | 2000 mg/dL |
| Bilirubin | 66 mg/dL |
| Rheumatoid Factor | 1200 IU/mL |
| Human IgG | 4 g/dL |
| Human IgM | 0.7 g/dL |
| Human IgA | 1.6 g/dL |
The sponsor states the following limitation in the labeling:
"Samples showing visible signs of hemolysis may cause interference. Avoid analyzing samples with hemoglobin concentrations $>0.2\mathrm{g/L}$."
"Samples should not be taken from patients receiving therapy with high biotin doses (i.e. $>5\mathrm{mg/day}$) until at least 8 hours following the last biotin administration."
HAMA Effect: The effect of the presence of human anti-mouse antibodies on the Elecsys Calcitonin assay was assessed on the cobas e 411 Immunoassay Analyzer. A high HAMA human serum pool $805~\mu\mathrm{g/L}$ $(0.805~\mu\mathrm{g/mL})$ was spiked with analyte to yield calcitonin concentrations of approx. 10 and $500~\mathrm{pg/mL}$. The high HAMA serum pool was diluted in 11 dilution series with a serum pool without HAMA. The expected analyte concentrations were calculated and compared with the observed concentrations for each diluted sample. The sponsor defined significant interference of HAMA as greater than $\pm10\%$ of the expected concentrations. All results were within $\pm10\%$ of the expected concentrations.
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The sponsor states the following limitation in the labeling:
"Samples from patients routinely exposed to animal or animal serum products may contain heterophilic antibodies causing an atypical result. This assay has been formulated to mitigate the risk of this type of interference. However, potential interactions between rare sera and test components can occur."
Drug Interference: To assess drug interference on the measurement of calcitonin, 17 pharmaceutical compounds were spiked into two human serum sample pools and tested with the Elecsys Calcitonin assay on the cobas e 411 Immunoassay analyzer. The analyte concentration of the sample pools were approx. 10 and 500 pg/mL Calcitonin (spiked). The sponsor defines significant interference as greater than $\pm 10\%$ of the reference value (unspiked sample). The drug interference results are summarized in the table below:
| Drug | Highest concentration of drug tested which showed no significant interference (mg/L) | Drug | Highest concentration of drug tested which showed no significant interference (mg/L) |
| --- | --- | --- | --- |
| Acetylcystein | 1700 | Levothyroxine | 1 |
| Ampicillin-Na | 1000 | Carbimazol | 30 |
| Ascorbic acid | 300 | Thiamazol | 80 |
| Cyclosporine | 5 | Propylthiouracil | 60 |
| Cefoxitin | 2500 | Sodium Perchlorate | 2000 |
| Heparin | 5000 U | Propranolol | 240 |
| Intralipid | 10000 | Amiodaron | 200 |
| Levodopa | 20 | Prednisolone | 100 |
| Methyldopa +1.5 | 20 | Hydrocortisone | 200 |
| Metronidazole | 200 | Fluocortolon | 100 |
| Phenylbutazone | 400 | Octreotid | 0.3 |
| Doxycycline | 50 | Iodide | 400 |
| Acetylsalicylic acid | 1000 | Theophylline | 100 |
| Rifampicin | 65 | Ibuprofen | 500 |
| Acetaminophen | 200 | | |
f. Assay cut-off: Not applicable
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2. Comparison studies:
a. Method comparison with predicate device:
A method comparison study was performed comparing the Siemens Immulite 1000 Calcitonin assay run on the Immulite 1000 analyzer (predicate device) to the Elecsys Calcitonin assay (candidate device) run on the cobas e 411 analyzer. A total of 150 human serum samples were measured in singlicate over a minimum of 5 days. Of those samples, 19 (12.7%) were altered in order to cover the entire measuring range. Calcitonin values ranged between 1.61 and 1936 pg/mL for the samples tested. Results of the Deming Regression analysis are presented below:
Deming Regression: y = 0.998x - 3.05, r = 0.991
95% CI of the slope: 0.967/1.03
95% CI of the intercept: -0.452/-0.159
b. Matrix comparison:
The effect on quantitation of calcitonin in the presence of anticoagulants with the Elecsys Calcitonin Immunoassay was determined by comparing values obtained from samples drawn into Li-Heparin tubes with and without serum separator gel, K₂ EDTA and K₃ EDTA plasma tubes. Fifty serum/plasma pairs per sample material were tested in singleton with one reagent lot on one cobas e 411 Immunoassay analyzer. The Passing/Bablok regression analysis results are summarized below:
| Serum vs | n | Slope | Intercept | Range (pg/mL) |
| --- | --- | --- | --- | --- |
| Lithium Heparin | 53 | 0.951 | 0.282 | 1.65-1911 |
| Li-Heparin Gel Separation | 54 | 0.955 | 0.176 | 1.65-1911 |
| K2-EDTA | 52 | 0.956 | 0.129 | 1.65-1911 |
| K3-EDTA | 53 | 0.958 | 0.080 | 1.65-1911 |
The resulting data support the package insert claim that serum, Li-Heparin plasma from tubes with and without gel separation, K₂-EDTA plasma, and K₃-EDTA-plasma are acceptable sample types for use with the Elecsys Calcitonin Immunoassay.
3. Clinical studies:
a. Clinical Sensitivity: Not applicable
{14}
b. Clinical specificity: Not applicable
c. Other clinical supportive data (when a. and b. are not applicable): Not applicable
4. Clinical cut-off: Not applicable
5. Expected values/Reference range:
Reference ranges for apparently health males (n=184) and apparently healthy females (n=180) were determined using the 97.5th percentiles as upper limit of normal. The evaluation was done at three different sites with two reagent lots using three cobas e 411 analyzers. The reference range study result summary is presented below:
| Cohort | N | 97.5th percentile | Lower limit of 95% CI | Upper limit of 95% CI |
| --- | --- | --- | --- | --- |
| Apparently healthy females | 180 | 7.63 pg/mL | 6.10 pg/mL | 12.7 pg/mL |
| Apparently healthy males | 184 | 14.3 pg/mL | 10.4pg/mL | 18.0 pg/mL |
N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
