(56 days)
Ortho-mune OKB19A PE Conjugate is intended for use in identification and enumeration of CD19+ human B lymphocytes in whole blood by flow cytometry.
Ortho-mune OKB19A Monoclonal Antibody (Murine) Phycoerythrin (PE) Conjugate contains the purified monoclonal antibody OKB19A conjugated to the fluorochrome phycoerythrin.
The provided document describes the performance characteristics of the Ortho-mune™ OKB™19A (CD19) Monoclonal Antibody (Murine) Phycoerythrin Conjugate, a diagnostic device for identifying and enumerating CD19+ human B lymphocytes.
Here's an analysis of the acceptance criteria and the study that proves the device meets them:
1. Table of Acceptance Criteria and Reported Device Performance:
The document doesn't explicitly state numerical acceptance criteria in a formal table. Instead, the acceptance is based on demonstrating "substantial equivalence" to a predicate device (Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE (Phycoerythrin)]). The performance is assessed through various studies. Below is a table summarizing the reported device performance and implicitly derived acceptance criteria based on equivalence:
| Performance Metric | Acceptance Criteria (Implicit, based on equivalence to predicate) | Reported Device Performance (Ortho-mune OKB19A) |
|---|---|---|
| Comparative Performance (Normal Donors) | Mean % CD19+ and Range % CD19+ should be comparable to the predicate device. | Mean % CD19+: 13.3, Range % CD19+: 1.3 - 31.1 |
| Comparative Performance (AIDS/ARC Patients) | Mean % CD19+ and Range % CD19+ should be comparable to the predicate device. | Mean % CD19+: 10.2, Range % CD19+: 0.4 - 41.9 |
| Linear Regression (Relative to Predicate) | High correlation (R value close to 1) and a slope close to 1, indicating similar measurements. | R = 0.909, Equation: Y = 0.711 + 0.88(X) (for combined data) |
| Within-Laboratory Reproducibility (Normal) | Coefficient of Variation (CV) should be acceptable for flow cytometry assays, similar to predicate performance (not explicitly stated, but assumed to be low). | Mean % Positive: 15.330, Site A CV: 5.082, Site B CV: 4.635, Site C CV: 4.545 |
| Within-Laboratory Reproducibility (Low) | CV should be acceptable. | Mean % Positive: 1.134, Site A CV: 24.182, Site B CV: 20.793, Site C CV: 22.106 |
| Within-Laboratory Reproducibility (High) | CV should be acceptable. | Mean % Positive: 26.760, Site A CV: 3.438, Site B CV: 3.401, Site C CV: 3.447 |
| Between-Laboratory Reproducibility (Normal) | Low CV across sites. | Overall Across Site CV: 0.976 |
| Between-Laboratory Reproducibility (Low) | Low CV across sites. | Overall Across Site CV: 1.466 |
| Between-Laboratory Reproducibility (High) | Low CV across sites. | Overall Across Site CV: 0.782 |
| Linearity (across lymphocyte count range) | Slopes indistinguishable from 1 and R values close to 1.000. | Slopes between 1.001 and 1.004, R values of 1.000 (for each donor and all combined) |
2. Sample Sizes Used for the Test Set and Data Provenance:
- Comparative Performance Study (against predicate):
- Sample Size: 205 normal donors and 87 AIDS/ARC patients.
- Data Provenance: The study was conducted at "three external, geographically distinct sites." The specific countries are not mentioned, but the distributed nature suggests a multi-center study. The data is prospective as specimens were stained and analyzed during the study.
- Reproducibility Studies:
- Sample Size: 10 normal donors for each of the three independent laboratories.
- Data Provenance: Three independent laboratories (suggesting different locations, likely within the same country as the submitter, USA, but not explicitly stated). The data is prospective as samples were processed and stained for the study.
- Linearity Study:
- Sample Size: Four normal donors.
- Data Provenance: Not explicitly stated where the donors or testing took place, but implied as part of the overall development and testing by the submitter. The data is prospective.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts:
This information is not applicable to this device. The device is a diagnostic reagent for flow cytometry, which measures cell populations based on antibody binding. The "ground truth" for the comparative study is the measurement obtained by the predicate device and the clinical classification of the patients (normal donor vs. AIDS/ARC). For reproducibility and linearity, the ground truth is statistical consistency and expected values determined by dilutions and other analytical methods, not expert consensus on an image or clinical case.
4. Adjudication Method for the Test Set:
This information is not applicable. Adjudication methods (like 2+1, 3+1) are typically used in studies where human readers are making subjective diagnoses or classifications from images/data, and discrepancies need to be resolved. This study involves objective measurements from a flow cytometer.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:
No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. MRMC studies are associated with evaluating the effectiveness of diagnostic tools where multiple human readers interpret cases. This study is comparing a new reagent's performance to an existing reagent using instrumental measurements.
6. Standalone (Algorithm Only Without Human-in-the-Loop Performance) Study:
Yes, the studies described are essentially a standalone performance evaluation of the Ortho-mune OKB19A reagent. The flow cytometer analyzes the stained cells, and the resulting data (percentage of CD19+ cells) is the output of the "algorithm" (the staining and detection process). Human involvement is in sample preparation, running the instrument, and data analysis, but the core measurement itself is automated and doesn't involve human interpretation of complex patterns in the way an AI algorithm for image analysis would. The performance is assessed purely on the device's ability to accurately identify and enumerate the target cells, comparing its output to a predicate device's output.
7. Type of Ground Truth Used:
- Comparative Performance: The ground truth was effectively the measurement obtained by the predicate device (Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE (Phycoerythrin)]) on the same samples. The clinical categorisation of "normal donor" and "AIDS/ARC patient" also serves as a clinical context for evaluating performance.
- Reproducibility Studies: The ground truth was based on the statistical expectation of consistent measurements when the same samples are tested multiple times (within-laboratory) and across different laboratories (between-laboratory). The "true" value for the percentage of CD19+ cells in the prepared samples was the reference point.
- Linearity Study: The ground truth was the expected values calculated by multiplying serial dilutions by the hematology analyzer-derived buffy coat lymphocyte count and the CYTORONABSOLUTE-derived lymphocyte subset percent positive.
8. Sample Size for the Training Set:
This information is not applicable as this is not an AI/Machine Learning device that requires a training set. The device is a monoclonal antibody conjugate; its performance is based on its biochemical properties and interaction with target cells.
9. How the Ground Truth for the Training Set Was Established:
This information is not applicable for the reason stated above.
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JAN 2 2 1997
REVISED 510(k) SUMMARY
SUBMITTER:
Ortho Diagnostic Systems 1001 U.S. Hwy 202 Raritan, NJ 08869-0606
CONTACT:
Joanne Harris Tel: (908) 218-8404 Fax: (908) 218-8168
DEVICE NAME:
Ortho-mune™ OKB™19A (CD19) Monoclonal Antibody (Murine) Phycoerythrin Conjugate
PREDICATE:
Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE Phycoerythrin)
DATE:
May 14, 1996
DEVICE DESCRIPTION
Ortho-mune OKB19A Monoclonal Antibody (Murine) Phycoerythrin (PE) Conjugate contains the purified monoclonal antibody OKB19A conjugated to the fluorochrome phycoerythrin.
INTENDED USE
Ortho-mune OKB19A PE Conjugate is intended for use in identification and enumeration of CD19+ human B lymphocytes in whole blood by flow cytometry. The intended use is the same as the intended use of the predicate device, Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE (Phycoerythrin)] commercially distributed by Becton Dickinson Immunocytometry Systems.
TECHNOLOGICAL CHARACTERISTICS
Both Ortho-mune OKB19A Monoclonal Antibody (Murine) PE Conjugate and Monoclonal Antibody Test for B Cells utilize monoclonal antibodies specific for human B cells (OKB19A/Leu-12) respectively, conjugated to the same fluorochrome, phycoerythrin.
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PERFORMANCE CHARACTERISTICS
Performance of Ortho-mune OKB19A Monocional Antibody (Murine) PE Conjugate was compared with that of Monoclonal Antibody Test For B Cells at three external, geographically distinct sites. Whole blood specimens from 205 normal donors, and 87 AIDS/ARC patients were stained and analyzed using the ORTHO CYTORONABSOLUTE™ flow cytometer, Ortho Diagnostic Systems Inc.
For each specimen, the percentage of gated cells which showed positive by each marker was calculated. The mean and range of the percent CD19+ cells for the normal donor and AIDS/ARC population are shown in Table 1 and Table 2 respectively.
TABLE 1
| PERCENT POSITIVE STAINED CELLS IN NORMAL DONORS DETECTED BYOKB19A AND LEU-12 ASSAYED ON THE CYTORONABSOLUTEN=205 | |||||
|---|---|---|---|---|---|
| Ortho-muneReagent | Mean % | Range % | BDReagent | Mean % | Range % |
| OKB19A(CD19)+ | 13.3 | 1.3 - 31.1 | LEU-12(CD19)+ | 14.1 | 0.1 - 31.3 |
TABLE 2
| PERCENT POSITIVE STAINED CELLS IN AIDS/ARC PATIENTS DETECTED BYOKB19A AND LEU-12 ASSAYED ON THE CYTORONABSOLUTEN=87 | |||||
|---|---|---|---|---|---|
| Ortho-muneReagent | Mean % | Range % | BDReagent | Mean % | Range % |
| OKB19A(CD19)+ | 10.2 | 0.4 - 41.9 | LEU-12(CD19)+ | 11.1 | 0.8 - 38.7 |
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Linear regression analysis of total percent CD19+ cells from the combined normal and AIDS/ARC populations is found in Chart 1.
Image /page/2/Figure/2 description: The image is a scatter plot titled "Ortho-mune OKB19A(PE)". The x-axis is labeled "Predicate Reagent", and the y-axis is labeled "Ortho-mune Reagent". The plot shows a positive correlation between the two reagents. The equation of the line of best fit is Y = 0.711 + 0.88(X), and the R-value is 0.909.
Ortho-mune OKB19A (CD19) PE vs Monocloual Autibody Test for B Cells (Leu-12)
This study demonstrates that the performance of Ortho-mune OKB19A (CD19) Monoclonal Antibody (Murine) PE Conjugate is equivalent to Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE (Phycoerythrin)] reagent in identification and enumeration CD19+ human lymphocytes in whole blood by flow cytometry.
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Reproducibility studies were performed at three independent laboratories using samples with low, normal, and high relative percent CD19+ cells.
Specimens from each of ten normal donors (whole blood, EDTA) were processed using monoclonal antibodies bound to microbeads to produce samples of low, normal, and high relative percent CD19+ cells. The samples were separated into aliquots for each laboratory. Samples were stained in replicates of 10 with Ortho-mune OKB19A. (CD19) PE Conjugate reagent and analyzed using the ORTHO CYTORONABSOLUTE flow cytometer.
For within laboratory reproducibility, the variance for the replicate results was calculated within site, concentration and donor. The variance was averaged across site, concentration and donor. The square root replicate variance (SD) was divided by the appropriate mean percent positive result (by site and concentration) and multiplied by 100 to obtain the CV. Within laboratory reproducibility results for determination of total percent CD19+ cells are presented in Table 3.
TABLE 3
| Concerner Color Color N=10 donors N=10 donors - N=10 donors - 10 donors - 10 done | HHUN LAHa martin min 2018 12:22 2 : 9 : 9 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 : 2 | OF A FOR VIREPRODUCIBILITY | |||||
|---|---|---|---|---|---|---|---|
| ortho-mune OKB19A(PF) | ALESTICSSMeanPercentPositive | Site ACV | A # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # # #Rens I | Site Rite Rite Real | CV Rens CV Ren | ||
| TOTAL CD19 Low CareerTOTAL CD19' Normal | 1.13415.330 | 24.1825.082 | 100100 | 20.7934.635 | 10098 | 22.1064.545 | 100100 |
| TOTAL CD19 High | 26.760 | 3.438 | ਰੇਰੇ | 3.401 | 100 | 3.447 | 100 |
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The between laboratory CV was computed as follows. The mean percent positive for The between faboratory of This computed. The SD was computed on the three site means within concentration was on the CV was obtained by dividing the SD by the overall means within concentration and multiplying by 100. Between laboratory reproducibility mean within consembly of total percent CD19+ cells are presented in Table 4.
| 6.4 | ••• | ||
|---|---|---|---|
| C | and the | C |
| BETWEEN LABORATORY REPRODUCIBILITY | |||||
|---|---|---|---|---|---|
| Ortho-mune OKB19A(PE) | |||||
| N=10 donors | |||||
| SITE A | SITE B | SITE C | ACROSS SITE | ||
| Ortho-mune OKB19A(PE) | Mean Percent Positive (All Donors) | Mean Percent Positive (All Donors) | Mean Percent Positive (All Donors) | Coefficient of Variation | # Reps |
| TOTAL CD19+ Low | 1.132 | 1.152 | 1.119 | 1.466 | 300 |
| TOTAL CD19+ Normal | 15.395 | 15.410 | 15.144 | 0.976 | 298 |
| TOTAL CD19+ High | 26.518 | 26.787 | 26.930 | 0.782 | 299 |
Ortho-mune OKB19A (CD19) Monoclonal Antibody (Murine) PE Conjugate immunophenotyping reagent shows acceptable within and between laboratory reproducibility for determination of CD19+ lymphocyte percentages
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A linearity study was performed using an automated hematology analyzer to determine total lymphocyte count, and the CYTORONABSOLUTE flow cytometer to determine the percent positive CDx cells.
Specimens from four normal donors (whole blood, EDTA) were processed to produce samples with low, normal and high numbers of lymphocyte subsets. Each whole blood specimen was concentrated by harvesting the buffy coat to obtain a white blood cell count between 20,000 and 40,000 cells/uL and then diluting to produce samples of high, normal and low numbers of lymphocyte subsets. A portion of each sample was stained in triplicate using Ortho-mune OKB19A (CD19) PE Conjugate immunophenotyping reagent and analyzed using the CYTORONABSOLUTE flow cytometer. The total lymphocyte count of the concentrated sample for each donor was obtained using an automated hematology analyzer.
Linear regression analyses were performed as follows. The expected (X axis ) values were calculated by multiplying the corresponding serial dilutions by the hematology analyzer derived buffy coat lymphocyte count and by the CYTORONABSOLUTE derived Jymphocyte subset percent positive. The observed (Y axis) values were determined as the total lymphocyte count calculated from the hematology derived value of the concentrated sample times the CYTORONABSOLUTE derived lymphocyte subset percent positive at each dilution.
The Ortho-mune OKB19A (CD19) PE Conjugate reagent demonstrated linear performance for total CD19+ lymphocyte subsets across a lymphocyte count range of 20 cells/uL to 9000 cells/uL as demonstrated with slopes indistinguishable from 1 and R values of 1.000.
Linear regression analyses of observed versus expected values for total percent CD19+ cells for each donor specimen are shown in. Regression analysis statistics are provided in Table 5.
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CHART 2
Image /page/6/Figure/1 description: The image is a scatter plot with the x-axis labeled "Expected CD18+ Counts" and the y-axis labeled "Observed CD18+ Counts". The plot shows a strong positive correlation between the expected and observed counts, with data points clustered tightly around a diagonal line. The x and y axis range from 0 to 1400. There are several data points plotted on the graph.
Ortho'-mune OKB19A(PE)
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| 0)All can a diffeA(C511 1119.10 | 11Children |
|---|---|
| ------------------------------------------------------------- | -------------------- |
| LINEARITY | ||||||
|---|---|---|---|---|---|---|
| Ortho-mune OKB19A(PE) | ||||||
| N=4 | ||||||
| Ortho-muneOKB19A(PE) | Donor | SLOPE | CI | INTERCEPT | CI | R |
| TOTAL CD19+ | 1 | 1.003 | 0.01 | -6.012 | 5.33 | 1.000 |
| TOTAL CD19+ | 2 | 1.004 | 0.00 | -4.017 | 2.59 | 1.000 |
| TOTAL CD19+ | 3 | 1.003 | 0.00 | -4.126 | 2.20 | 1.000 |
| TOTAL CD19+ | 4 | 1.001 | 0.00 | 0.311 | 1.36 | 1.000 |
| TOTAL CD19+ | All | 1.002 | 0.00 | -3.308 | 1.50 | 1.000 |
CONCLUSION
Performance of Ortho-mune OKB19A (CD19) Monoclonal Antibody (Murine) Phycoerythrin Conjugate is substantially equivalent to Monoclonal Antibody Test for B Cells [Anti-Leu-12 (CD19) PE (Phycoerythrin)] in the identification and enumeration of CD19+ human T lymphocytes in whole blood by flow cytometry.
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Reproducibility studies were performed at three independent laboratories using samples with low, normal, and high relative percent CD19+ cells.
Specimens from each of ten normal donors (whole blood, EDTA) were processed using monoclonal antibodies bound to microbeads to produce samples of low, normal, and high relative percent CD19+ cells. The samples were separated into aliquots for each laboratory. Samples were stained in replicates of 10 with Ortho-mune OKB19A (CD19) PE Coniugate reagent and analyzed using the ORTHO CYTORONABSOLUTE flow cytometer.
For within laboratory reproducibility, the variance for the replicate results was calculated within site, concentration and donor. The variance was averaged across site, concentration and donor. The square root replicate variance (SD) was divided by the appropriate mean percent positive result (by site and concentration) and multiplied by 100 to obtain the CV. Within laboratory reproducibility results for determination of total percent CD19+ cells are presented in Table 3.
TABLE 3
| WITHIN LABORATORY REPRODUCIBILITY | |||||||
|---|---|---|---|---|---|---|---|
| Ortho-mune OKB19A(PE) | |||||||
| N = 10 donors | |||||||
| Ortho-mune OKB19A(PE) | All SITESMeanPercentPositive | Site A | Site B | Site C | |||
| TOTAL CD19+ Low | 1.134 | 24.182 | 100 | 20.793 | 100 | 22.106 | 100 |
| TOTAL CD19+ Normal | 15.330 | 5.082 | 100 | 4.635 | 98 | 4.545 | 100 |
| TOTAL CD19+ High | 26.760 | 3.438 | 99 | 3.401 | 100 | 3.447 | 100 |
§ 864.5220 Automated differential cell counter.
(a)
Identification. An automated differential cell counter is a device used to identify one or more of the formed elements of the blood. The device may also have the capability to flag, count, or classify immature or abnormal hematopoietic cells of the blood, bone marrow, or other body fluids. These devices may combine an electronic particle counting method, optical method, or a flow cytometric method utilizing monoclonal CD (cluster designation) markers. The device includes accessory CD markers.(b)
Classification. Class II (special controls). The special control for this device is the FDA document entitled “Class II Special Controls Guidance Document: Premarket Notifications for Automated Differential Cell Counters for Immature or Abnormal Blood Cells; Final Guidance for Industry and FDA.”