(33 days)
In vitro diagnostic test for the quantitative determination of the HDL-cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.
A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.
The HDL-Cholesterol Gen.4 is a homogeneous enzymatic colorimetric test. Non-HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water-soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non-HDL lipoproteins is blocked. Finally only HDL-particles can react with CHER and CHOD. The concentration of HDL-cholesterol is determined enzymatically by CHER and CHOD.
Here's a breakdown of the acceptance criteria and study details for the HDL-Cholesterol Gen.4 device, extracted from the provided document:
Device: HDL-Cholesterol Gen.4
Type: In vitro diagnostic test for the quantitative determination of HDL-cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.
Acceptance Criteria and Reported Device Performance
| Acceptance Criteria Category | Specific Criteria (Implicitly met by study design and results) | Reported Device Performance |
|---|---|---|
| Precision | Low %CV for repeatability and intermediate precision across various HDL-C concentrations. | Repeatability: - PreciControl ClinChem Multi 1 (Mean 28.0 mg/dL): SD 0.20 mg/dL, CV 0.7% - PreciControl ClinChem Multi 2 (Mean 68.1 mg/dL): SD 0.44 mg/dL, CV 0.6% - Human Serum 1 (Mean 9.48 mg/dL): SD 0.17 mg/dL, CV 1.8% - Human Serum 2 (Mean 40.5 mg/dL): SD 0.26 mg/dL, CV 0.7% - Human Serum 3 (Mean 59.4 mg/dL): SD 0.32 mg/dL, CV 0.5% - Human Serum 4 (Mean 79.4 mg/dL): SD 0.51 mg/dL, CV 0.6% - Human Serum 5 (Mean 141 mg/dL): SD 0.83 mg/dL, CV 0.6%Intermediate Precision: - PreciControl ClinChem Multi 1 (Mean 28.4 mg/dL): SD 0.30 mg/dL, CV 1.1% - PreciControl ClinChem Multi 2 (Mean 66.4 mg/dL): SD 0.9 mg/dL, CV 1.4% - Human Serum 1 (Mean 9.48 mg/dL): SD 0.20 mg/dL, CV 2.2% - Human Serum 2 (Mean 40.7 mg/dL): SD 0.33 mg/dL, CV 0.8% - Human Serum 3 (Mean 59.4 mg/dL): SD 0.40 mg/dL, CV 0.7% - Human Serum 4 (Mean 79.4 mg/dL): SD 0.65 mg/dL, CV 0.8% - Human Serum 5 (Mean 141 mg/dL): SD 1.07 mg/dL, CV 0.8% |
| Analytical Sensitivity (LoB, LoD, LoQ) | LoB, LoD, and LoQ should be below the claimed measuring range. | LoB Observed: 0.00 mg/dL (Claim: 3.09 mg/dL)LoD Observed: 0.50 mg/dL (Claim: 3.09 mg/dL)LoQ Observed: 2.89 mg/dL (Claim: 3.09 mg/dL) |
| Linearity/Assay Reportable Range | Measurements should be linear across the claimed measuring range (3.09 to 150 mg/dL), with good correlation and low deviation. | Serum: Slope 1.020, Intercept -0.399, Correlation Coefficient (r2) 0.9992, Repeatability 1.5%Plasma: Slope 1.022, Intercept -0.173, Correlation Coefficient (r2) 0.9929, Repeatability 0.8%Claimed Measuring Range: 3.09 to 150 mg/dL (for both serum and plasma).Nonlinearity did not deviate by more than 10%. |
| Endogenous Interferences | No significant interference (bias >10%) from common interferents like hemolysis, lipemia, icterus, and triglycerides at specified levels. | Hemolysis: No Interference up to 1200 H indexLipemia: No Interference up to 2000 L indexUnconjugated Bilirubin: No Interference up to 60 I indexConjugated Bilirubin: No Interference up to 60 I indexTriglycerides: No Interference up to 1200 mg/dL |
| Exogenous Interferences (Drugs) | No significant interference from a panel of common drugs at specified concentrations. | Results provided in Table 7 (specific details of "no interference" for each drug are implied by listing them in a "Test Concentrations Results" table within the interference section, assuming they met the 10% bias criterion for non-interference). |
| Method Comparison to Predicate | Strong correlation with a legally marketed predicate device, with acceptable bias at medical decision points. | Regression Analysis (HDL-Cholesterol Gen.4 vs. Predicate): y = 0.956x - 0.949, r = 0.995Bias at medical decision points: -6.7 % at 40.2 mg/dL -6.0 % at 59.9 mg/dL |
| Matrix Comparison | Acceptable correlation between serum and various plasma anticoagulant types. | Serum vs. Serum Gel Separation: y = 0.99x - 0.33, r = 0.999Serum vs. Li-heparin: y = 0.99x - 0.32, r = 1.000Serum vs. K2-EDTA: y = 0.98x - 0.70, r = 0.999Serum vs. K3-EDTA: y = 0.95x - 0.08, r = 0.999 |
Study Details
-
Sample sizes used for the test set and the data provenance:
- Precision (Repeatability & Intermediate Precision): 5 human serum pools and 2 control samples were used. Tested for 21 days with 4 replicates/day. Data provenance is not explicitly stated as country of origin, but implied to be from a laboratory setting (likely in the US or Germany, given Roche's locations). The nature of the study (analyzing human serum pools and controls) suggests prospective data collection for evaluating device performance.
- Analytical Sensitivity (LoB, LoD, LoQ):
- LoB: One analyte-free sample. Measured 10-fold in 6 runs over 3 days, per reagent lot (total 60 measurements per lot).
- LoD: Five samples with low-analyte concentration. Measured two-fold in 6 runs over 3 days, per reagent lot (total 60 measurements per lot).
- LoQ: A low-level sample set prepared by diluting 5 human serum samples. Tested in 5 replicates per sample on 5 days.
- Linearity/Assay Reportable Range: Dilution series prepared using 1 serum pool and 1 plasma pool. The dilution series contained 11 concentrations for serum and 15 concentrations for plasma.
- Endogenous Interferences: Two human serum pools spiked with HDL-Cholesterol and interfering substances.
- Exogenous Interferences (Drugs): Two human serum sample pools.
- Method Comparison to Predicate: 111 routine laboratory serum samples. Additionally, 4 samples spiked with high human serum HDL-Cholesterol and 1 sample diluted with 0.9% NaCl. Data provenance is not explicitly stated as country of origin, but implied to be from a laboratory setting. The use of "routine laboratory serum samples" suggests retrospective collection, though the spiking and dilution aspects are prospective for the study design.
- Matrix Comparison: 38 paired samples (serum and plasma) from single donors.
-
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- For this type of in vitro diagnostic device (quantitative measurement of a biomarker), "ground truth" is typically established by reference methods or validated comparative methods, not by human expert consensus or clinical adjudication as would be seen in imaging diagnostics.
- The predicate device (Ultra N-geneous HDL Cholesterol Reagent, K021316) serves as a comparative "ground truth" for the method comparison study. The precision, sensitivity, linearity, and interference studies establish the intrinsic performance properties of the device against predefined analytical standards (e.g., CLSI guidelines).
- Therefore, the concept of "experts" as in "a radiologist with 10 years of experience" is not directly applicable here. The experts involved would be laboratory scientists, biochemists, and statisticians who designed and executed the studies according to CLSI guidelines and interpreted the analytical data.
-
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- Adjudication methods like 2+1 or 3+1 are typically used in clinical imaging studies where subjective interpretation is involved.
- For this in vitro diagnostic device, measurements are quantitative, and "adjudication" is done through statistical analysis and adherence to predefined acceptance criteria based on established analytical guidelines (e.g., CLSI EP5-A3 for precision, CLSI EP17-A2 for detection limits, CLSI EP6-A for linearity). There is no "human adjudication" process for individual results as there would be for image interpretation.
-
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:
- No, an MRMC comparative effectiveness study was not done. This type of study is relevant for imaging devices where human readers interpret medical images, and AI might assist or replace them. The HDL-Cholesterol Gen.4 is an in vitro diagnostic device that quantifies a substance in a laboratory sample; it does not involve human "readers" interpreting results in a subjective or visual manner that AI would enhance.
-
If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- Yes, the performance evaluation reports are inherently demonstrating the "standalone" performance of the analytical algorithm/reagent system. The studies evaluate the device's ability to accurately and precisely measure HDL-cholesterol concentrations in samples, independent of further human interpretation beyond routine laboratory operation and quality control. The reported results (e.g., mean, SD, CV, regression equations) directly reflect the algorithm's performance.
-
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- Analytical Ground Truth: For the precision, sensitivity, linearity, and interference studies, the "ground truth" is based on the known concentrations of analytes in prepared samples (e.g., spiked samples, diluted samples, control materials) or the absence of analytes (blank samples), and performance is evaluated against established analytical standards and acceptable deviations.
- Comparative Ground Truth: For the method comparison study, the predicate device (Ultra N-geneous HDL Cholesterol Reagent, K021316) served as the comparative "ground truth" or reference method for evaluating substantial equivalence. This is a common approach for 510(k) clearances.
- No pathology or outcomes data was used for establishing the ground truth for device performance in this submission, as it's an analytical performance study for an IVD.
-
The sample size for the training set:
- This document describes the pre-market notification (510(k)) studies for device validation, not the development or training of an AI algorithm. Therefore, there is no "training set" for an AI mentioned or implied in this submission. The device is a chemical reagent system, not an AI/ML independent medical device.
-
How the ground truth for the training set was established:
- As noted above, no "training set" for an AI algorithm is described in this submission. The ground truth for the analytical studies described (e.g., precision, linearity) is based on the preparation of samples with known concentrations or comparative analysis against a validated predicate device, as per standard laboratory practice and CLSI guidelines.
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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
ROCHE DIAGNOSTICS OPERATIONS (RDO) BARBARA MCWHORTER REGULATORY AFFAIRS PRINCIPAL 9115 HAGUE ROAD SUITE 114 INDIANAPOLIS IN 46250
October 19, 2016
Re: K162593
Trade/Device Name: HDL-Cholesterol Gen.4 Regulation Number: 21 CFR 862.1475 Regulation Name: Lipoprotein test system Regulatory Class: I, meets the limitation of exemption 21 CFR §862.9(c)(4) Product Code: LBS Dated: September 14, 2016 Received: September 16, 2016
Dear Ms. McWhorter:
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 (OS) 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/ResourcesforYou/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, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K162593
Device Name HDL-Cholesterol Gen.4
Indications for Use (Describe)
In vitro diagnostic test for the quantitative determination of the HDL-cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.
A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.
| 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) |
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HDL-Cholesterol Gen.4 510(k) Summary
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).
The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the HDL-Cholesterol Gen.4 reagent.
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| Submitter Name | Roche Diagnostics |
|---|---|
| Address | 9115 Hague RoadP.O. Box 50416Indianapolis, IN 46250-0457 |
| Contact | Barbara McWhorterPhone: (317) 521-2336FAX: (317) 521-2324Email: barbara.mcwhorter@roche.com |
| Date Prepared | September 13, 2016 |
| Proprietary Name | HDL-Cholesterol Gen.4 |
| Common Name | HDL-Cholesterol |
| Classification Name | Lipoprotein test system |
| Product Codes,Regulation Numbers | LBS, 21 CFR § 862.1475, Class I, meets the limitations to the exemptions 21CFR§ 862.9(c)(4) |
| Predicate Devices | Ultra N-geneous HDL Cholesterol Reagent, K021316 |
| Establishment Registration | For the HDL-Cholesterol Gen.4, the establishment registration numberfor Roche Diagnostics GmbH in Mannheim, Germany is 9610126. Theestablishment registration number for Roche Diagnostics in the UnitedStates is 1823260. |
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1. DEVICE DESCRIPTION
The HDL-Cholesterol Gen.4 is a homogeneous enzymatic colorimetric test. Non-HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water-soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non-HDL lipoproteins is blocked. Finally only HDL-particles can react with CHER and CHOD. The concentration of HDL-cholesterol is determined enzymatically by CHER and CHOD.
2. INDICATIONS FOR USE
In vitro diagnostic test for the quantitative determination of the HDL-cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.
A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.
TECHNOLOGICAL CHARACTERISTICS 3.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER. In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4-cholestenone and hydrogen peroxide. In the presence of peroxidase, the hydrogen peroxide generated reacts with 4 -amino-antipyrine and EMSE to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.
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Figure 1: Test Principle
CHER ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------HDL-cholesterol esters + Hoo In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4-cholestenone and hydrogen peroxide. CHOD ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------HDL-cholesterol + O2 A4-cholestenone + H2O2 In the presence of peroxidase, the hydrogen peroxide generated reacts with 4-amino-antipyrine and EMSE®) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically. Peroxidase 2 H₂O₂ + 4-amino-antipyrine + —————— colored pigment + 5 H2O EMSEp) + H+ + H2O
The following table compares the HDL-Cholesterol Gen.4 with its predicate device, Ultra Ngeneous HDL Cholesterol Reagent (K021316).
Table 1: Assay Comparison
| Feature | Predicate DeviceUltra N-geneous HDL Cholesterol Reagent,(K021316) | Candidate DeviceHDL-Cholesterol Gen.4 |
|---|---|---|
| Intended Use | For the quantitative measurement of high-density lipoprotein cholesterol(HDL-C) concentration in human serum orplasma. | In vitro diagnostic test for the quantitativedetermination of theHDL-cholesterol concentration in humanserum and plasma onRoche/Hitachi cobas c systems. |
| Feature | Predicate DeviceUltra N-geneous HDL Cholesterol Reagent,(K021316) | Candidate DeviceHDL-Cholesterol Gen.4 |
| Test Principle | The method is in a two reagent format anddepends on the properties of a uniquedetergent, as illustrated. This method is basedon accelerating the reaction of cholesteroloxidase (CO) with non-HDL unesterifiedcholesterol and dissolving HDL selectivelyusing a specific detergent. In the first reagent,non-HDL unesterified cholesterol is subject toan enzyme reaction and the peroxidegenerated is consumed by a peroxidasereaction with DSBmT yielding a colorlessproduct. The second reagent consists of adetergent capable of solubilizing HDLspecifically, cholesterol esterase (CE) andchromogenic coupler to develop color for thequantitative determination of HDL-C. This maybe referred to as the Accelerator SelectiveDetergent methodology. | Cholesterol esters are broken downquantitatively into free cholesterol and fattyacids by CHER. In the presence of oxygen,cholesterol is oxidized by cholesterol oxidaseto Δ4-cholestenone and hydrogen peroxide.In the presence of peroxidase, the hydrogenperoxide generated reacts with 4-amino-antipyrine and EMSE to form a dye. The colorintensity of this dye is directly proportional tothe cholesterol concentration and is measuredphotometrically. |
| ReagentComposition | R1:Cholesterol oxidase, (Fr: E. Coli),Peroxidase,(Fr: Horseradish), N,N-bis(4-sulphobutyl)-mtoluidine-disodium(DSBmT), Accelerator,Preservative, Ascorbic Oxidase,(Fr: Curcubitasp.)R2:Buffer, Cholesterol esterase (Fr:Pseudomonassp.), 4-Aminoantipyrine (4-AAP), Detergent,Preservative | R1:TAPSO buffer: 62.1 mmol/L, pH 7.77;polyanion: 1.25 g/L; EMSE: 1.08 mmol/L;ascorbate oxidase (cucurbita): ≥ 50 µkat/L;peroxidase (horseradish): ≥ 166.7 µkat/L ;detergent; BSA: 2.0 g/L; preservativeR2:Bis-Tris buffer: 20.1 mmol/L, pH 6.70;cholesterol esterase (microorganism): ≥ 7.5µkat/L; cholesterol oxidase (recombinant E.coli): ≥ 7.17 µkat/L; cholesterol oxidase(microorganism): ≥ 76.7 µkat/L; peroxidase(horseradish): ≥ 333 µkat/L; 4-amino-antipyrine: 1.48 mmol/L; BSA: 3.0 g/L;detergents; preservative |
| SampleType/Matrix | SerumPlasma: EDTA or lithium or sodiumheparin. | Serum.Plasma: Li-heparin, K2- and K3-EDTA plasma |
| Calibrator | Sekisui Diagnostics Ultra N-geneous® HDLCholesterol Calibrator or the HDL UltraCholesterol Calibrator | S1: H2OS2: C.f.a.s. Lipids |
| Feature | Predicate DeviceUltra N-geneous HDL Cholesterol Reagent,(K021316) | Candidate DeviceHDL-Cholesterol Gen.4 |
| Controls | The National Cholesterol Education Program(NCEP) Lipid Standardization Panel (LSP)recommends two levels of controls, one in thenormal range (40-65 mg/dL) and one near theconcentrations for decision making (<40mg/dL). An acceptable range of HDLcholesterol values should be established byeach laboratory. If control values are not withinthe expected range, confirm that procedureswere performed correctly and follow normaltroubleshooting measures. | PreciControl ClinChem Multi 1PreciControl ClinChem Multi 2 |
| Reagent Stability | Reagent 1 is stable open on the analyzer for 4weeks at 2-8°C.Reagent 2 is stable open on the analyzer for 4weeks at 2-8°C.DO NOT FREEZE | Shelf life at 2-8 °C: See expiration date oncobas c pack label.On-board in use and refrigerated on theanalyzer:12 weeks |
| Measuring Range | 2.5 mg/dL to 200 mg/dL | 3.09-150 mg/dL |
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NON-CLINICAL PERFORMANCE EVALUATION 4.
The following performance data were provided in support of the substantial equivalence determination:
Precision according to CLSI EP5-A3
Detection Limit: LoB, LoD and LoQ according to CLSI EP17-A2
Linearity according to CLSI EP6-A
Endogenous Interferences - H, L and I Indices
Endogenous Interferences - Triglycerides
Exogenous Interferences - Drugs
Method Comparison to Predicate
Matrix Comparison - Anticoagulants
4.1. Precision
Repeatability and Intermediate Precision 4.1.1.
Precision was evaluated per a protocol with modifications from the study design recommended in CLSI Guideline EP5-A3. The testing was run for 21 days; 4 individual replicates were tested in one run per day. Two replicates were tested at the beginning of the run, followed by 10 patient samples and then two replicates were tested at the end of the run. Five human serum pools and two control samples were tested on one cobas c 501, using 3 lots of reagent. Mean, Repeatability (within run precision) and Intermediate precision (within-lab precision) %CV and SD values were calculated.
Results and Conclusions 4.1.2.
| Specimen | Mean (mg/dL) | SD (mg/dL) | CV (%) |
|---|---|---|---|
| PreciControl ClinChem Multi 1 | 28.0 | 0.20 | 0.7 |
| PreciControl ClinChem Multi 2 | 68.1 | 0.44 | 0.6 |
| Human Serum 1 | 9.48 | 0.17 | 1.8 |
Table 2: Repeatability Summary
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| Specimen | Mean (mg/dL) | SD (mg/dL) | CV (%) |
|---|---|---|---|
| Human Serum 2 | 40.5 | 0.26 | 0.7 |
| Human Serum 3 | 59.4 | 0.32 | 0.5 |
| Human Serum 4 | 79.4 | 0.51 | 0.6 |
| Human Serum 5 | 141 | 0.83 | 0.6 |
Table 3: Intermediate Precision Summary
| Specimen | Mean (mg/dL) | SD (mg/dL) | CV (%) |
|---|---|---|---|
| PreciControl ClinChem Multi 1 | 28.4 | 0.30 | 1.1 |
| PreciControl ClinChem Multi 2 | 66.4 | 0.9 | 1.4 |
| Human Serum 1 | 9.48 | 0.20 | 2.2 |
| Human Serum 2 | 40.7 | 0.33 | 0.8 |
| Human Serum 3 | 59.4 | 0.40 | 0.7 |
| Human Serum 4 | 79.4 | 0.65 | 0.8 |
| Human Serum 5 | 141 | 1.07 | 0.8 |
Analytical Sensitivity 4.2.
4.2.1. Limit of Blank (LoB)
LoB of the HDL-Cholesterol Gen.4 on the cobas c 501 analyzer was determined according to CLSI EP17-A2. Limit of Blank determines the highest observed measurement values for samples free of analyte. The Limit of Blank was determined as the 95th percentile of measurements of blank samples.
One analyte free sample was measured on three lots of reagent in 10-fold determinations in 6 runs, distributed over 3 days, on one analyzer. In total, 60 measurements were obtained per reagent lot. Data analysis was based on determination of the 95th percentile of the 60 measured values.
LoB Observed: 0.00 mg/dL
LoB Claim: 3.09 mg/dL
Limit of Detection (LoD) 4.2.2.
LoD of the HDL-Cholesterol Gen.4 on the cobas c 501 analyzer was determined according to CLSI EP17-A2. The LoD determines the lower limit for samples with analyte. The LoD is the concentration at which there is a 95% probability that a sample contains analyte.
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For determination of LoD, five samples with low-analyte concentration (approximately up to 4 times the LoB) were measured with three lots of reagent in two-fold determination in 6 runs, distributed over 3 days, on one analyzer. In total 60 measurements were obtained per lot of reagent.
LoD was determined using the following equation:
LOD = LOB + 1.653 x SDtot
Where:
SDeeta = Square root [0.2 x ((SDamble )2 + (SD sample 2 + (SD sample 2 + (SD sample 2) + (SD sample 2 )].
LoD Observed: 0.50 mg/dL
LoD Claim: 3.09 mg/dL
Limit of Quantitation (LoQ) 4.2.3.
The LoQ of the HDL-Cholesterol Gen.4 was determined on the cobas c 501 analyzer according to CLSI Guideline EP17-A2.
The LoO is derived from a plot (mean concentration versus %CV ) with the goal of %CV less than or equal to 20% which was met. A low level sample set was prepared by diluting 5 human serum samples with an analyte free diluent (0.9% NaCl). The low level sample set was tested in 5 replicates per sample on 5 days, one run per day on one analyzer.
LoQ Observed: 2.89 mg/dL
LoQ Claim: 3.09 mg/dL
Linearity/Assay Reportable Range 4.3.
Regression Analysis 4.3.1. -
The linearity study was conducted to demonstrate that measurements across the claimed measuring range for each parameter are linear. The study was performed according to CLSI guideline EP6-A.
Dilution series were prepared using the human sample pools (1 serum pool and 1 plasma pool) with HDL-C concentrations above the upper end of the measuring range. Dilutions were made using 0.9% NaCl. The dilution series contain 11 concentrations for serum and 15 concentrations
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for plasma. Sample dilution levels were measured in triplicate and data analysis was done separately for each sample type.
Linear regression analysis was done according to EP6-A.
In a first step, a linearity check was performed with first order (linear) regression and then with higher order models (quadratic and cubic). A linearity check was performed with a first order (linear) regression for plasma and additionally a 2nd order regression for serum. The linear regression was weighted 1/Conc2.
The nonlinearity of the assay did not deviate by more than 10% during testing.
Table 4: Linearity Results
| Slope | Intercept | CorrelationCoefficient (r2) | Repeatability | ClaimedMeasuringRange | |
|---|---|---|---|---|---|
| Serum | 1.020 | -0.399 | 0.9992 | 1.5% | 3.09 to 150 mg/dL |
| Plasma | 1.022 | -0.173 | 0.9929 | 0.8% | 3.09 to 150 mg/dL |
Endogenous Interferences 4.4.
4.4.1. L. H. and I Indices
The effect on quantitation of analyte in the presence of endogenous interfering substances was determined at two HDL-Cholesterol concentrations (approximately 33 and 65 mg/dL) and a dilution set of the added interfering substances. Interfering substances evaluated include:
| Hemolysis | up to an H index of 1200 |
|---|---|
| Lipemia | up to an L index of 2000 |
| Icterus/Conjugated and Unconjugated Bilirubin | up to an I index of 60 |
High concentrated stock solutions of the interference substances were prepared in a suitable solvent. Two human serum pools were spiked with the defined HDL-Cholesterol concentrations and divided into two aliquots. The potential interfering substance was added to one aliquot, while the other aliquot was mixed with the same amount of solvent without the interfering substance. A
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dilution series was prepared with 11 dilution steps for each interferent by mixing the 2 aliquots. Three aliquots per level were tested in 1 run on 1 instrument and 1 lot.
The aliquot containing the interfering substance had the same HDL-Cholesterol concentrations as the aliquots containing no interfering substance. When diluting those two aliquots the HDL-Cholesterol concentration remained constant while the concentration of interferent varied. Thus the effect of increasing concentrations of interferent can be determined.
Mean of the measured results were compared to the expected result (aliquot with no interfering substance) and the recovery was determined (paired difference testing). Interference was defined as bias greater than 10%.
This procedure was repeated for each of the interfering substances.
| Interferent | Claim |
|---|---|
| Hemolysis | No Interference up to 1200 H index |
| Lipemia | No Interference up to 2000 L index |
| Unconjugated Bilirubin | No Interference up to 60 I index |
| Conjugated Bilirubin | No Interference up to 60 I index |
Table 5: H, L, I Indices/Serum Indices Interference
Triglycerides Interference 4.4.2.
The effect on quantitation of analyte in the presence of endogenous interfering substances was determined at two HDL-Cholesterol concentrations (approximately 33 and 65 mg/dL) and a dilution set of the added interfering substance.
High concentrated stock solution of triglycerides was prepared in a suitable solvent. Two human serum pools were spiked at the defined HDL-Cholesterol concentrations and divided into two aliquots. The potential interfering substance was added to one aliquot, while the other aliquot was mixed with the same amount of solvent without the interfering substance. A dilution series was prepared with 11 dilution steps for each interferent by mixing the 2 aliquots. Three aliquots per level were tested in 1 run on 1 instrument and 1 lot.
The parts containing the interfering substance had the same HDL-Cholesterol concentrations as the aliquots containing no interfering substance. When diluting those two aliquots the HDL-
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Cholesterol concentration remained constant while the concentration of triglycerides varied. Thus the effect of increasing concentrations of interferent can be determined.
Mean of the measured results were compared to the expected result (aliquot with no interfering substance) and the recovery was determined (paired difference testing).
Interference was defined as bias greater than 10%.
Table 6 Triglycerides Interference
| Interferent | Claim |
|---|---|
| Triglycerides | No Interference up to 1200mg/dLTriglycerides |
Exogenous Interferences – Drugs 4.5.
Two human serum sample pools spiked with approximately 30 and 60 mg/dL HDL-Cholesterol concentrations were divided into two aliquots. One aliquot of each concentration were used as the reference sample for HDL-Cholesterol concentration and were not spiked with the drugs but the solvent for the drug.
The other aliquots, with either the high or low HDL-Cholesterol concentration, were spiked with the respective amount of drug. The HDL-Cholesterol concentration of the spiked aliquots were tested with 3 replicates in one run, 1 reagent lot and one instrument. The defined drug compounds were spiked into samples with concentrations according to EP7-A2 or higher concentrations. The drugs included:
| Table 7: Potentially Interfering Drugs and Test Concentrations Results | |||
|---|---|---|---|
| Potential Interferent | Concentrationmg/L |
|---|---|
| Acetylcystein | 553 |
| Ampicillin-Na | 1000 |
| Ascorbic acid | 500 |
| Cyclosporine | 5 |
| Cefoxitin | 2500 |
| Heparin | 5000 U |
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| Potential Interferent | Concentration | mg/L |
|---|---|---|
| Intralipid | 10000 | |
| Levodopa | 4 | |
| Methyldopa +1.5 | 4 | |
| Metronidazole | 200 | |
| Phenylbutazone | 400 | |
| Doxycyclin | 50 | |
| Acetylsalicylic Acid | 1000 | |
| Rifampicin | 60 | |
| Acetaminophen | 200 | |
| Ibuprofen | 500 | |
| Theophyllin | 100 | |
| Bezafibrate | 120 | |
| Simvastatin | 16 |
Method Comparison to Predicate 4.6.
One hundred eleven routine laboratory serum samples were used in the method comparison testing. No patient information was obtained.
Four samples were spiked with high human serum HDL-Cholesterol. One sample was diluted with 0.9% NaCl.
The samples were tested in singlicate using the HDL Ultra Cholesterol reagent from Sekisui on Roche/Hitachi 917 analyzer and the HDL-Cholesterol Gen.4 reagent on cobas c 501 analyzer.
The data was evaluated using Passing Bablok Regression analysis.
Regression analysis results:
y = 0.956x - 0.949, r = 0.995
Bias at medical decision points:
-6.7 % at 40.2 mg/dL
- -6.0 % at 59.9 mg/dL
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4.7. Matrix Comparison - Anticoagulants
Each pair of serum and plasma of a single donor were spiked with HDL-Cholesterol. The 38 paired sample tubes are tested in singlicate.
A method comparison was executed by using the serum data as the reference. Only samples within the measuring range were used.
The following method comparisons were completed:
Gel separation tubes
K2-EDTA plasma vs serum
K3-EDTA plasma vs serum
Li-Heparin plasma vs serum
Table 8: Matrix Comparison Results
| Anticoagulant | Linear Regression | Range Tested |
|---|---|---|
| Serum vs. Serum Gel Separation | y = 0.99x - 0.33, r = 0.999 | 3.48 to 145 mg/dL |
| Serum vs. Li-heparin | y = 0.99x - 0.32, r = 1.000 | 3.48 to 147 mg/dL |
| Serum vs. K2-EDTA | y = 0.98x - 0.70, r = 0.999 | 3.87 to 144 mg/dL |
| Serum vs. K3-EDTA | y = 0.95x - 0.08, r = 0.999 | 3.87 to 145 mg/dL |
CLINICAL PERFORMANCE EVALUATION 5.
Not Applicable
6. ADDITIONAL INFORMATION
Other Devices Required But Not Provided 6.1.
- The Calibrator f.a.s. Lipids, K011658 .
- The PreciControl ClinChem Multi 1 and 2, K102016 .
- Diluent NaCl 9% .
There have been no changes to these items marketed with the new HDL-Cholesterol Gen.4.
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CONCLUSIONS 7.
The submitted information in this premarket notification supports a substantial equivalence decision.
§ 862.1475 Lipoprotein test system.
(a)
Identification. A lipoprotein test system is a device intended to measure lipoprotein in serum and plasma. Lipoprotein measurements are used in the diagnosis and treatment of lipid disorders (such as diabetes mellitus), atherosclerosis, and various liver and renal diseases.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 862.9.