(35 days)
The AquaLite® FSH Bioluminescent Immunoassay (BIA) Kit (or the AquaLite® FSH assay) is intended to be used in clinical laboratories for the quantitative determination of human FSH in sera and plasma. The AquaLite® FSH assay is for in vitro diagnostic use.
The AquaLite® FSH Bioluminescent Immunoassay Kit uses a polyclonal anti-FSH antibody that is pre-coated onto polystyrene tubes (solid phase). Samples (serum or plasma) and appropriate calibrators or controls, are pipetted (25 µL) into the pre-coated tubes. Anti-FSH Conjugate (150 uL) is then added to the tubes. The conjugate uses the photoprotein. AquaLite® (recombinant aequorin; Patent Nos. 5, 422, 266 and 5, 486, 455) which is covalently linked to an anti-FSH monoclonal antibody. FSH in the sample simultaneously combines with polyclonal antibody on the solid phase and conjugate antibody to form an immune complex or "sandwich" bound to the solid phase. Complex formation is complete after a 60-minute incubation period at room temperature (18°C to 25°C) on a standard orbital shaker. The tubes are then washed to remove unbound conjugate. The washed tubes are placed in a luminometer that is capable of reading a triggered, flash-type reaction in 12 x 75 mm tubes. An injected calcium trigger solution causes AquaLite® to oxide its self-contained luciferin molecule. This reaction produces a flash of light at 469 nm. which is measured by the luminometer. The intensity of the light is directly proportional to the concentration of the FSH in the sample. To calculate results, the light intensity (in relative light units, RLU) of the FSH calibrators is plotted against FSH concentration (in International Units per liter. IU/L) to vield a calibration curve. This curve is used to relate the light intensity generated from the samples and controls to FSH concentration in IU/L.
The information provided is for an in vitro diagnostic test (AquaLite® FSH Bioluminescent Immunoassay (BIA) Kit), not a medical device in the typical sense of AI-powered systems analyzed for clinical improvement. Therefore, many of the requested fields, particularly those related to AI, multi-reader multi-case studies, and ground truth established by expert consensus or pathology, are not applicable.
However, I can extract and present the acceptance criteria and the study results for the performance characteristics of this immunoassay as an analogy to the requested format.
Here’s an interpretation of the provided 510(k) summary into the requested structure:
Acceptance Criteria and Device Performance for SeaLite Sciences, Inc. AquaLite® FSH
1. Table of Acceptance Criteria and Reported Device Performance
| Performance Characteristic | Acceptance Criteria (Implied) | Reported Device Performance |
|---|---|---|
| Sensitivity | Low detection limit for FSH | 0.03 IU/L |
| Specificity | Low cross-reactivity with related hormones (hCG, LH, TSH) | hCG: 0.02%, LH: 0.10%, TSH: 1.90% |
| High Dose Hook Effect | No hook effect at clinically relevant high concentrations | No high dose hook effect occurs prior to 4,000 IU/L FSH. |
| Precision (Intra-assay) | Low % CV across a range of FSH concentrations (e.g., <10-15%) | 6.69 IU/L: 5.4% CV26.89 IU/L: 6.65% CV45.28 IU/L: 7.15% CV |
| Precision (Inter-assay) | Low % CV across a range of FSH concentrations (e.g., <10-15%) | (For similar levels as intra-assay, based on context)*7.52%, 6.39%, 4.049% |
| Method Comparison | High correlation with a commercially available kit (e.g., R > 0.9) | Correlation coefficient: 0.90 (Slope: 0.66, y-intercept: 0.78) |
| Linearity and Nonparallelism | Consistent recovery across dilutions (e.g., 90-110%) | Recovery range: 93% to 118% |
| Spike and Recovery | Recovery within an acceptable range (e.g., 90-110%) | Recovery range: 100% to 128% (Most within 100-115%) |
| Recovery in Serum and Plasma | No significant differences across different sample types | No significant differences among serum and SST serum, or heparin, EDTA, oxalate, and citrate plasmas. Recoveries mostly 90-109% compared to standard serum. |
| Effect of Common Interferents | No significant effect on FSH quantification by common interferents | No significant effect by hemoglobin, bilirubin, human serum albumin, or triglycerides at tested levels. Recoveries: 88-105% (for 0 spike), 94-104% (for 25.7 spike). |
*Note on Inter-assay precision: The table formatting for inter-assay precision is corrupted in the input document, but the provided %CV values indicate similar low variability to intra-assay (7.52%, 6.39%, and 4.049%).
2. Sample size used for the test set and the data provenance
- Sensitivity: 20 replicates of a zero-level calibrator.
- Specificity: Not explicitly stated as a "test set" in terms of patient samples. Tested using aliquots of WHO/NIBSC preparations of purified hormones.
- Precision (Intra-assay): N = 10 per solution for each of three FSH levels (total 30 samples).
- Precision (Inter-assay): N = 20 per solution (2 replicates x 10 assays) for three FSH levels (total 60 samples).
- Method Comparison: N = 92 patient samples.
- Linearity and Nonparallelism: 5 human serum samples.
- Spike and Recovery: 8 normal human serum samples.
- Recovery in Serum and Plasma: Blood samples from 2 normal subjects, processed into 6 different sample types each.
- Effect of Common Interferents: Pooled normal male human serum.
Data Provenance: The studies were conducted at SeaLite Sciences, Inc. The samples appear to be clinical samples (patient samples, human serum), but the specific country of origin is not explicitly stated, although WHO/NIBSC (London, England) reference materials were used for specificity. The data is retrospective in the sense that the experiments were conducted and then analyzed, but not in the context of analyzing pre-existing patient data for an algorithm.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
Not applicable. This is an immunoassay, and the "ground truth" for the test set (e.g., FSH concentration, absence of interferents, hormone specificity) is established by the known characteristics of the samples or the reference measurements, not by expert interpretation.
4. Adjudication method for the test set
Not applicable. As an immunoassay, the results are quantitative measurements, not subjective evaluations requiring adjudication.
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 immunoassay, not an AI-powered diagnostic system involving human readers.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, in so far as an immunoassay can be considered "standalone." The device (AquaLite® FSH assay) provides quantitative results directly from the sample without human interpretation of complex images or signals requiring an 'algorithm' in the AI sense. Its performance characteristics (sensitivity, specificity, precision, etc.) are measured as direct outputs of the device.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)
The "ground truth" for this immunoassay is primarily based on:
- Known concentrations/compositions of reference materials: For sensitivity (zero calibrator), specificity (purified hormones), linearity (dilution of samples with known FSH), spike-and-recovery (known amount of spiked FSH).
- Reference standard methods/devices: For method comparison (comparison to a "commercially available kit").
- Biological realism: For assessing recovery in different matrices (serum/plasma) and the effect of interferents.
8. The sample size for the training set
Not applicable. This is a biochemical assay, not an machine learning algorithm that requires a discrete training set. The assay's parameters (e.g., antibody concentrations, incubation times) are developed through R&D, but not in the same way an AI model is "trained."
9. How the ground truth for the training set was established
Not applicable (see point 8). The assay's performance is optimized through traditional experimental design and chemical/biological principles during its development phase.
{0}------------------------------------------------
SeaLite Sciences. Inc.
510(K) SUMMARY
FEB 1 4 1997
GENERAL INFORMATION I.
Trade or proprietary name - SeaLite Sciences, Inc. AquaLite® FSH
Common or usual name - Bioluminescent immunoassay (BIA)
Classification name - FDA has classified FSH test systems intended for the measurement of FSH in the diagnosis of pituitary gland and gonadal function.
Submitter's Name and Address: Cathryn N. Cambria Director Regulatory Affairs and Quality Assurance, SeaLite Sciences, Inc. 3000 Northwoods Parkway, Suite 200 Norcross. Georgia 30071 (800) 874-4471, extension 227 Submission Date: January 9, 1997
Legally Marketed Device to which Claim Substantial Equivalence: SeaLite Sciences, Inc. AquaLite® FSH Assay
DEVICE DESCRIPTION II.
The AquaLite® FSH Bioluminescent Immunoassay Kit uses a polyclonal anti-FSH antibody that is pre-coated onto polystyrene tubes (solid phase). Samples (serum or plasma) and appropriate calibrators or controls, are pipetted (25 µL) into the pre-coated tubes. Anti-FSH Conjugate (150 uL) is then added to the tubes. The conjugate uses the photoprotein. AquaLite® (recombinant aequorin; Patent Nos. 5, 422, 266 and 5, 486, 455) which is covalently linked to an anti-FSH monoclonal antibody. FSH in the sample simultaneously combines with polyclonal antibody on the solid phase and conjugate antibody to form an immune complex or "sandwich" bound to the solid phase. Complex formation is complete after a 60-minute incubation period at room temperature (18°C to 25°C) on a standard orbital shaker. The tubes are then washed to remove unbound conjugate.
386422.1
{1}------------------------------------------------
SeaLite Sciences. Inc.
The washed tubes are placed in a luminometer that is capable of reading a triggered, flash-type reaction in 12 x 75 mm tubes. An injected calcium trigger solution causes AquaLite® to oxide its self-contained luciferin molecule. This reaction produces a flash of light at 469 nm. which is measured by the luminometer. The intensity of the light is directly proportional to the concentration of the FSH in the sample. To calculate results, the light intensity (in relative light units, RLU) of the FSH calibrators is plotted against FSH concentration (in International Units per liter. IU/L) to vield a calibration curve. This curve is used to relate the light intensity generated from the samples and controls to FSH concentration in IU/L. Note that the numerical value for FSH in mIU/mL is the same as for IU/L (International System). For example, 15.6 mIU/mL equals 15.6 IU/L.
Note: Samples that generate signals greater than the signal from the highest calibrator These samples must be diluted with Calibrator A and re-assayed. are off-scale. Remember to multiply the results from diluted samples by the dilution factor used.
III. SUMMARY OF STUDIES AND TECHNOLOGICAL CHARACTERISTICS
Studies on SeaLite Sciences, Inc. AquaLite® FSH were conducted at SeaLite Sciences. The results are summarized below:
Performance Characteristics
1. Sensitivity
The sensitivity or detection limit of the AquaLite® FSH assay is 0.03 IU/L. Sensitivity is determined by adding the mean signal of twenty (20) replicates of the zero level calibrator plus two (2) standard deviations above this mean. The FSH concentration (IU/L) corresponding to this calculated signal is defined as the analytical sensitivity of the assay.
2. Specificity
The AquaLite® FSH assay measures intact FSH. The following human sialoglycoprotein hormones were supplied by the World Health Organization's National Institute for Biological Standards and Controls (London, England). Aliquots of these preparations were diluted to the following levels in zero calibrator and assayed. Percent cross-reactivity (%) is reported below:
{2}------------------------------------------------
SeaLite Sciences, Inc.
| Substance | WHO/NIBSCLot Number | Tested at | % Cross-reactivity |
|---|---|---|---|
| hCG | 3rd IS 75/537 | 2,500 IU/L | 0.02 |
| LH | 2nd IS 80/552 | 1,000 IU/L | 0.10 |
| TSH | 2nd IRP 80/558 | 1,000 mIU/L | 1.90 |
- High Dose Hook Effect No high dose hook effect occurs prior to 4,000 3. IU/L FSH.
Precision 4.
- Intra-assay precision. Tri-level commercial controls containing FSH at (a) the following concentrations were assayed to determine intra-assay precision. (Total N = 10 per solution.)
| Mean FSH Level(IU/L) | SD | % CV(calibration values) |
|---|---|---|
| 6.69 | 0.36 | 5.4 |
| 26.89 | 1.80 | 6.65 |
| 45.28 | 3.23 | 7.15 |
- Inter-assay precision. Tri-level commercial controls containing FSH at (b) the following concentrations were assayed in duplicate repetitively. Ten assays were performed using ten sets of calibration values. Interassay precision observed for the solutions (Total N = 2 x 10 = 20) is shown below.
| SD | % CV |
|---|---|
| 0.485 | 7.52 |
| 1.679 | 6.39 |
| 4.049 | તે જેવી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ |
{3}------------------------------------------------
Method Comparison 5.
The AquaLite® FSH assay was used to test patient samples (N=92) that were previously assayed by a commercially available kit. The samples ranged from 1.4 to 230 IU/L. A slope of 0.66 with a y-intercept of 0.78 was obtained. The correlation coefficient was 0.90.
Linearity and Nonparallelism 6.
Five human serum samples containing the levels of endogenous FSH shown below were diluted in parallel using Calibrator A.
| SampleID | DilutionFactor | Found(IU/L) | Expected(IU/L) | Recovery(%) |
|---|---|---|---|---|
| 2 | Undiluted1:21:41:8 | 179.692.142.124.2 | --89.944.922.45 | --10294108 |
| 3 | Undiluted1:21:41:8 | 104.848.829.412.7 | --52.426.213.1 | --9310997.6 |
| 6 | Undiluted1:21:41:8 | 54.124.514.98.1 | --2713.56.8 | --90.7110115 |
| 30 | Undiluted1:21:41:8 | 134.675.136.119.7 | --67.333.616.8 | --111106113 |
| 42 | Undiluted1:21:41:8 | 163.584.341.426.1 | --81.754120.5 | --102101118 |
{4}------------------------------------------------
7. Spike and Recovery
Eight normal human serum samples were spiked with 25 IU/L FSH using WHO FSH (2nd IRP 78/549). The spiked samples were assayed using the AquaLite® FSH assay. All values are in IU/L.
| Sample ID | Unspiked | FSHMeasured | FSHExpected | %Recovery |
|---|---|---|---|---|
| 1 | 4.2 | 29.2 | 29.2 | 100 |
| 12 | 4.4 | 34.3 | 29.4 | 115 |
| 14 | 7.3 | 37.4 | 32.3 | 115 |
| 17 | 4.9 | 32.6 | 29.9 | 109 |
| 20 | 7.1 | 35.5 | 32.1 | 110 |
| 54 | 7.9 | 42.4 | 32.9 | 128 |
| 57 | 7.4 | 35.1 | 32.4 | 108 |
| 81 | 7.3 | 37.2 | 32.3 | 115 |
8. Recovery in Serum and Plasma
Blood Samples from 2 normal subjects were prepared as sera (standard technique and SST tubes) as well as heparin, EDTA, oxalate, and citrate plasmas. FSH was quantified using the AquaLite® FSH assay. Recovered FSH was compared with FSH recovered in serum (standard technique). The data demonstrate that there are no significant differences among serum and SST serum nor among serum and heparin, EDTA, oxalate and citrate plasmas when using the AquaLite® FSH assay. All values are in IU/L.
| Sample | P1 | % | P2 | % |
|---|---|---|---|---|
| Serum | 9.35 | 100 | 9.59 | 100 |
| SST | 9.54 | 102 | 10.5 | 109 |
| EDTA | 9.07 | 97 | 8.62 | 90 |
| Heparin | 9.89 | 106 | 10.4 | 109 |
| Oxalate | 8.50 | 91 | 8.8 | 92 |
| Citrate | 7.75 | 83 | 9.1 | 95 |
{5}------------------------------------------------
SeaLite Sciences, Inc.
9. Effect of Common Interferents
Pooled normal male human serum was spiked with preparations of hemoglobin, bilirubin, human serum albumin and triglycerides to the levels shown below. Equal amounts of FSH were spiked into normal male serum as well as the normal male serum aliquots containing potential interferents. FSH was quantified using the AquaLite® FSH assay. Recovered FSH was compared with the FSH recovered in normal male serum. The data (in IU/L) demonstrate that the AquaLite® FSH assay is not significantly affected by hemoglobin, bilirubin, human serum albumin or triglycerides at the levels tested.
| FSH | Hemoglobin(at 500 mg/dL) | Bilirubin(at 20 mg/dL) | Albumin(at 12 mg/dL) | Triglycerides(at 3000 mg/dL) | |||||
|---|---|---|---|---|---|---|---|---|---|
| Spike | Serum | IU/L | % | IU/L | % | IU/L | % | IU/L | % |
| 0 | 9.77 | 9.83 | 100 | 9.13 | 93 | 10.2 | 105 | 8.56 | 88 |
| 25.7 | 35.5 | 36.3 | 102 | 35.6 | 94 | 37.0 | 104 | 36.6 | 103 |
CONCLUSIONS DRAWN FROM STUDIES IV.
The data from the studies conducted demonstrated that the performance of SeaLite Sciences, Inc. AquaLite® FSH is similar and substantially equivalent to that of other commercially available assays for FSH.
V. ALTERNATIVE PRACTICES AND PROCEDURES
There are several assay technologies commonly employed to measure the presence of human FSH in serum or plasma. They include: radioimmunoassay (RIA) and enzyme-linked immunosorbent immunoassay (ELISA).
POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH VI.
Use Universal Precautions. No known test method can offer complete assurance that products derived from human serum are pathogen-free; therefore, handle all materials of human origin as though they were potentially infectious.
Sodium azide is used as a preservative. This preservative may react with metallic plumbing to form explosive metal azides. Flush with large volumes of water when disposing of materials containing sodium azide.
{6}------------------------------------------------
As an in vitro diagnostic test, there are not direct adverse effects on the health of a patient from the use of this product. However, failure of the device to perform as indicated, the contamination of reagents, the use of reagents past the labeled expiration dates, the use of improper specimens, or human error during the performance of the test may lead to erroneous results and possible improper patient management.
VII. INDICATIONS FOR USE
The AquaLite® FSH Bioluminescent Immunoassay (BIA) Kit (or the AquaLite® FSH assay) is intended to be used in clinical laboratories for the quantitative determination of human FSH in sera and plasma. The AquaLite® FSH assay is for in vitro diagnostic use.
§ 862.1300 Follicle-stimulating hormone test system.
(a)
Identification. A follicle-stimulating hormone test system is a device intended to measure follicle-stimulating hormone (FSH) in plasma, serum, and urine. FSH measurements are used in the diagnosis and treatment of pituitary gland and gonadal disorders.(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.