(60 days)
Immunoassay for the in vitro quantitative determination of estradiol in human serum and plasma.
Competition principle. Total duration of assay: 18 minutes, 37 °C.
· 1st incubation (9 minutes): By incubating the sample (50 uL) with an estradiol-specific biotinylated antibody (65 uL), an immunocomplex is formed, the amount of which is dependent upon the analyte concentration in the sample.
·2nd incubation (9 minutes): After addition of streptavidin-coated microparticles (35 µL) and an estradiol derivative labeled with a ruthenium complex* (65 uL), the still-vacant sites of the biotinylated antibodies become occupied, with the formation of an antibody-hapten complex. The entire complex becomes bound to the solid phase via interaction of biotin and streptavidin.
· The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier (0.4 second read frame).
·Results are determined via a calibration curve which is instrumentspecifically generated by 2-point calibration and a master curve provided via the reagent bar code.
** Tris(2,2'-bipyridy))ruthenium(II) complex (Ru(bpy)" 3)
The provided text describes a 510(k) summary for the Boehringer Mannheim Elecsys® Estradiol Assay, comparing it to a predicate device, the Enzymun-Test® Estradiol Assay (K916132). This submission focuses on establishing substantial equivalence based on performance characteristics.
Here's an analysis of the acceptance criteria and the study that proves the device meets them:
1. A table of acceptance criteria and the reported device performance
The document does not explicitly state formal "acceptance criteria" with pass/fail thresholds. Instead, it presents performance characteristics of the new device (Elecsys® Estradiol) and directly compares them to the predicate device (Enzymun-Test® Estradiol). The implicit acceptance criterion is that the performance of the Elecsys® Estradiol Assay is comparable to or better than the predicate device.
Given this, I will construct a table comparing the performance of the Elecsys® Estradiol Assay with the Enzymun-Test® Estradiol Assay, as this constitutes the 'reported device performance' against an implicit 'acceptance' of being equivalent to the predicate.
| Feature | Elecsys® Estradiol Performance | Enzymun® Estradiol Performance (Predicate) | Implicit Acceptance Criteria (Comparison to Predicate) |
|---|---|---|---|
| Precision | Comparable precision (within-run and total) | ||
| Within-Run (Mean) | Low: 7.7, Mid: 44.0, High: 277.6, High: 1217.0 (pg/mL) | Low: 8.2, Mid: 53.3, High: 3021 (pg/mL) (Note: Different levels reported for Elecsys vs. Enzymun) | - |
| Within-Run (%CV) | Low: 3.0, Mid: 2.7, High: 3.3, High: 2.7 (Note: Different levels reported for Elecsys vs. Enzymun) | Low: 14.3, Mid: 16.1, High: 9.1 | - |
| Total (Mean) | Low: 9.0, Mid: 44.0, High: 277.6, High: 1217.0 (pg/mL) | Low: 10.3, Mid: 53.3, High: 3021 (pg/mL) | - |
| Total (%CV) | Low: 9.0, Mid: 9.0, High: 10.0, High: 10.0 (Note: Different levels reported for Elecsys vs. Enzymun) | Low: 16.1, Mid: 16.1, High: 10.3 | - |
| Lower Detection Limit | 10 pg/mL | 10 pg/mL | Equal or lower detection limit |
| Linearity | 10-4600 pg/mL (with a deviation from a linear line of ±10%) | 10-1300 pg/mL (with a deviation from a linear line of ±10%) | Comparable linearity range, potentially wider |
| Method Comparison (Vs Enzymun-Test Estradiol) | High correlation (r value > 0.97) and small bias | ||
| Least Squares | y = 1.027x + 6.32; r = 0.995; SEE = 31.03; N = 64 | y = 0.902x + 42.94; r = 0.976; SEE = 91.96; N = 74 | - |
| Passing/Bablok | - | y = 0.962x + 12.51; r = 0.976; SEE = 38.71; N = 74 | - |
| Interfering Substances (No interference at) | Bilirubin: 64.5 mg/dL; Hemoglobin: 1.0 g/dL; Lipemia: 1250 mg/dL; Biotin: 20 ng/mL | Bilirubin: 25 mg/dL; Hemoglobin: 200 mg/dL; Lipemia: 1500 mg/dL; Biotin: 24 ng/mL | Comparable or improved resistance to interference |
| Specificity (% Cross-reactivity) | Low cross-reactivity, comparable to predicate or better | ||
| 17 hydroxy-Progesterone | < 0.5 | < 0.001 | - |
| Ethisterone | < 0.5 | --- | - |
| Norethindrone-Acetate | < 0.5 | --- | - |
| 2-Methoxy-estradiol | < 1.0 | 0.07 | - |
| 6-Hydroxy-estradiol | < 600 | 62 | - |
| Estriol | < 1.0 | 0.3 | - |
| Prednisolone | < 0.25 | --- | - |
| Danazol | <0.1 | 0.0006 | - |
| Testosterone | <1.0 | 0.03 | - |
| Di-Hydro-Testosterone | < 1.0 | --- | - |
| 5-Androsten-3B-17B-Diol | < 0.4 | --- | - |
2. Sample sized used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)
- Sample Size for Method Comparison:
- Least Squares: N = 64
- Passing/Bablok: N = 74
- Precision: The "N" value for precision testing is 10 for each reported level (Low, Mid, High). This represents the number of replicates used to calculate the mean and %CV for precision for both within-run and total precision.
- Data Provenance: The document does not specify the country of origin of the data or whether the studies were retrospective or prospective.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)
This is a clinical chemistry assay, not an imaging device requiring expert interpretation. The "ground truth" for method comparison and performance evaluation in such assays is typically established by comparing the new method to a reference method (in this case, the predicate Enzymun-Test Estradiol Assay) or a more definitive method like ID-GC/MS for standardization.
- No experts (like radiologists) were used to establish ground truth in the context of human interpretation, as the device doesn't involve image analysis.
- Assay Standardization: The document mentions "Assay Standardization: ID-GC/MS." This indicates that Isotope Dilution Gas Chromatography/Mass Spectrometry (ID-GC/MS), a highly accurate and precise reference method, was used to standardize both the Elecsys® Estradiol Assay and the predicate Enzymun-Test® Estradiol Assay. While not explicitly called "ground truth" for the test set, it serves as the foundational "truth" for calibrating and establishing the accuracy of the assays.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set
Not applicable for this type of in vitro diagnostic device. Adjudication methods like 2+1 or 3+1 are typically used in studies involving human interpretation of medical images where disagreements among readers need to be resolved. This device directly measures a biomarker concentration.
5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
Not applicable. This is an automated in vitro diagnostic assay. It does not involve human readers interpreting images or AI assistance.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the studies presented (precision, linearity, method comparison, interfering substances, specificity) represent the standalone performance of the Elecsys® Estradiol Assay as an automated in vitro diagnostic device. There is no human-in-the-loop performance aspect beyond standard laboratory procedures for sample handling and operating the instrument.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The "ground truth" for the performance evaluation can be considered multifaceted:
- Method Comparison: The predicate device, Enzymun-Test® Estradiol Assay, served as the primary comparative "ground truth" for showing substantial equivalence.
- Assay Standardization: ID-GC/MS (Isotope Dilution Gas Chromatography/Mass Spectrometry) was used for establishing the foundational accuracy of the assay values, acting as a higher-level "ground truth" for calibration.
- Interference and Specificity: These are determined by spiking known concentrations of potential interferents/cross-reactants and observing their effect on the measured estradiol value, with the known absence of interference/cross-reactivity being the "ground truth."
8. The sample size for the training set
The document does not specify a "training set" in the context of machine learning. This is an assay based on established biochemical principles (immunoechemistry) and calibration rather than a machine learning algorithm that requires a separate training set. The calibration curve is generated by 2-point calibration and a master curve provided via the reagent bar code. This master curve and the 2-point calibration define the device's operational parameters, which could be loosely analogous to what a "training set" provides in AI, but it's not a direct comparison.
9. How the ground truth for the training set was established
As there is no distinct "training set" for a machine learning algorithm, the concept of "ground truth for the training set" as typically understood in AI/ML is not directly applicable.
However, if we consider the calibration of the assay as analogous to "training":
- The calibration curve is generated using calibrators of known estradiol concentrations. The "ground truth" for these calibrators would be established through careful quantitative analysis using highly accurate reference methods, again likely involving methods traceable to ID-GC/MS or similar definitive techniques. The reagent bar code also provides a "master curve," implying pre-determined, highly characterized calibration data.
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| 510(k) Summary | |
|---|---|
| Introduction | According to the requirements of 21 CFR 807.92, the following informationprovides sufficient detail to understand the basis for a determination ofsubstantial equivalence. |
| 1.Submittername,address,contact | Boehringer Mannheim Corporation2400 Bisso LaneP.O. Box 4117Concord, CA 94524-4117(510) 674 - 0690 extension 8413FAX (510) 687 - 1850Contact Person: Yvette LloydDate Prepared: December 13, 1996 |
| 2.Device name | Proprietary name: Elecsys® Estradiol AssayCommon name: Electrochemiluminescence assay for the determination ofestradiol.Classification name: System, Test, Estradiol |
| 3.Predicatedevice | We claim substantial equivalence to the Enzymun-Test® Estradiol Assay(K916132). |
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and the comments of the comments of
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- Device Description Competition principle. Total duration of assay: 18 minutes, 37 °C.
· 1st incubation (9 minutes): By incubating the sample (50 uL) with an estradiol-specific biotinylated antibody (65 uL), an immunocomplex is formed, the amount of which is dependent upon the analyte concentration in the sample.
·2nd incubation (9 minutes): After addition of streptavidin-coated microparticles (35 µL) and an estradiol derivative labeled with a ruthenium complex* (65 uL), the still-vacant sites of the biotinylated antibodies become occupied, with the formation of an antibody-hapten complex. The entire complex becomes bound to the solid phase via interaction of biotin and streptavidin.
· The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier (0.4 second read frame).
·Results are determined via a calibration curve which is instrumentspecifically generated by 2-point calibration and a master curve provided via the reagent bar code.
** Tris(2,2'-bipyridy))ruthenium(II) complex (Ru(bpy)" 3)
ડ. Intended use
Immunoassay for the in vitro quantitative determination of estradiol in human serum and plasma.
Continued on next page
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- Comparison to predicate device
The Boehringer Mannheim Elecsys® Estradiol Assay is substantially equivalent to other products in commercial distribution intended for similar use. Most notably it is substantially equivalent to the currently marketed Enzymun-Test® Estradiol Assay (K916132).
The following table compares the Elecsys® Estradiol Assay with the predicate device, Enzymun-Test® Estradiol Assay. Specific data on the performance of the test have been incorporated into the draft labeling in attachment 5. Labeling for the predicate device in provided in attachment 6.
Similarities:
•Intended Use: Immunoassay for the in vitro quantitative determination of estradiol
·Sample type: Serum and plasma
· Antibody: Same polyclonal anti-estradiol antibody
· Solid phase binding principle: Streptavidin/Biotin
· Assay Standardization: ID-GC/MS
: -
Differences:
ﺔ ﺍﻟﻘ
, i
| Feature | Elecsys® Estradiol | Enzymun® Estradiol |
|---|---|---|
| Reaction testprinciple | Electrochemiluminescence | ELISA/1-step sandwichassay. |
| Instrumentrequired | Elecsys® 2010 | ES 300 |
| CalibrationStability | A calibration isrecommended every 7 daysif kit is not consumed; 4weeks with same reagent lotif reagent is consumedwithin 7 days. | Full calibration requiredevery 2 weeks. One-pointcalibration required everyrun. |
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Performance Characteristics:
Comparison to predicate device, cont.
Enzymun® Estradiol Elecsys® Estradiol Feature Modified NCCLS (pg/mL): Modified NCCLS (pg/mL): Precision Mid High High Low Mid Low Level ୧୦ ୧୦ 60 ୧୦ ୧୦ N ୧୦ 1217.0 44.0 277.6 3021 ર 3.3 410 Within-Run: Mean 7.7 8 . ર 3.0 14.3 2.7 6.5 %CV 1217.0 277.6 3021 44.0 410 53.3 Total: Mean 9.1 10.3 ર રે 16.1 ર .0 %CV 9.0 10 pg/mL 10 pg/mL Lower Detection Limit 10-1300 pg/mL (with a 10-4600 pg/mL (with a Linearity deviation from a linear line deviation from a linear line ﺔ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤﺘﺤﺪﺓ ﺍﻟﻤ of ±10%) of ±10%) Vs Enzymun-Test Vs Enzymun-Test Method Estradiol Estradiol Comparison Least Squares Least Squares y = 1.027x + 6.32 y = 0.902x + 42.94 r = 0.995 r = 0.976 SEE = 31.03 SEE = 91.96 N = 64 N = 74 . Passing/Bablok y = 0.962x + 12.51 r = 0.976 SEE = 38.71 N = 74 No interference at: No interference at: Interfering substances 64.5 mg/dL 25 mg/dL Bilirubin 200 mg/dL 1.0 g/dL Hemoglobin 1250 mg/dL 1500 mg/dL Lipemia 20 ng/mL Biotin 24 ng/mL
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Performance Characteristics:
Comparison to predicate device, cont.
| Feature | Elecsys® Estradiol | Enzymun® Estradiol | ||
|---|---|---|---|---|
| Specificity | Level tested(ng/mL) | % Cross-reactivity | Level tested | % Cross-reactivity |
| 17 hydroxy-Progesterone | 100 | < 0.5 | --- | < 0.001 |
| Ethisterone | 100 | < 0.5 | --- | --- |
| Norethindrone-Acetate | 100 | < 0.5 | --- | --- |
| 2-Methoxy-estradiol | 100 | < 1.0 | --- | 0.07 |
| 6-Hydroxy-estradiol | 0.78 | < 600 | --- | 62 |
| Estriol | 160 | < 1.0 | --- | 0.3 |
| Prednisolone | 1000 | < 0.25 | --- | --- |
| Danazol | 100000 | <0.1 | --- | 0.0006 |
| Testosterone | 100000 | <1.0 | --- | 0.03 |
| Di-Hydro-Testosterone | 100000 | < 1.0 | --- | --- |
| 5-Androsten-3B-17B-Diol | 100000 | < 0.4 | --- | --- |
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§ 862.1260 Estradiol test system.
(a)
Identification. An estradiol test system is a device intended to measure estradiol, an estrogenic steroid, in plasma. Estradiol measurements are used in the diagnosis and treatment of various hormonal sexual disorders and in assessing placental function in complicated pregnancy.(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.