(90 days)
For In Vitro Diagnostic Use
Mouse Anti-Human T-cell, CD3/FITC, UCHT1 + Mouse Anti-Human T-cell, CD8/RPE, DK25 (DAKO Anti-CD3/FITC and Anti-CD8/RPE) has been developed for use in flow cytometry for the analysis of CD3+ and CD8+ T-cells. This reagent allows simultaneous detection and quantification of CD3+CD8+ cells (CD8 positive T-lymphocytes) in normal and pathological conditions such as immunodeficiency disorders. It is one component of the suggested monoclonal antibody (MAb) combination for routine immunophenotyping of lymphocytes in peripheral blood using flow cytometry.
Purified mouse anti-human CD3, Clone UCHT1, conjugated with fluorescein isothiocyanate, isomer 1 (FITC) + purified mouse anti-human CD8, Clone DK25, conjugated with R-phycoerythrin, present in 0.05M Tris-HCl buffer, pH 7.2, 15 mM NaN3, 0.1M NaCl, stabilized with 1% carrier protein
Subpopulations of lymphocytes may be stained with fluorochrome-conjugated antibody and evaluated in peripheral blood specimens when contaminating red blood cells (RBC's) are lysed prior to flow cytometric analysis. A subpopulation of WBC's are selected for assessment based upon cell morphology.
Here's a summary of the acceptance criteria and the study proving the device meets them, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance:
| Acceptance Criteria (Implicit) | Reported Device Performance |
|---|---|
| High correlation for measurement of CD3+ cells compared to predicate DAKO CD3/FITC. | The study showed a correlation greater than 0.99 for the measurement of CD3+ T-cells by DAKO Anti-CD3/FITC and Anti-CD8/RPE reagent compared to DAKO CD3/FITC (predicate device). |
| High correlation for measurement of CD8+ cells compared to predicate DAKO CD8/RPE. | The study showed a correlation greater than 0.98 for the measurement of CD8+ T-cells by DAKO Anti-CD3/FITC and Anti-CD8/RPE reagent compared to DAKO CD8/RPE (predicate device). |
| Linearity for CD3 detection. | Linearity testing of DAKO CD3/FITC (precursor to the combined reagent, and the CD3 component of the new device) using JM cells yielded the equation: y = 0.02 + 0.98x; with a correlation coefficient (r) of 0.999. (While this is presented for the single reagent, it supports the linearity of the CD3 component in the combined product). |
| Linearity for CD8 detection. | Linearity testing of DAKO CD8/RPE (precursor to the combined reagent, and the CD8 component of the new device) using JM cells yielded the equation: y = 0.06 + 1.01x; with a correlation coefficient (r) of 0.999. (Similar to CD3, this supports the linearity of the CD8 component). |
| Reproducibility of results (implied, as part of performance assessment). | Reproducibility of DAKO reagents (implied to include the combined reagent and/or its components) was measured using replicates run on two different flow cytometers at three concentrations of each antigen. The document states this testing was done, but does not provide specific performance metrics beyond stating the results indicate the new reagent performs as well as the predicates. |
| Minimal cross-reactivity with other peripheral blood cells (implied, as part of specificity). | Cross-reactivity of Anti-CD3/FITC plus Anti-CD8/RPE with peripheral blood cells (red blood cells, monocytes, granulocytes, lymphocytes, and platelets) was measured. The document states this testing was done, but does not provide specific performance metrics beyond stating the results indicated comparable performance. |
| Performance of combined reagent is comparable to individual predicate devices for detecting and enumerating CD3+ and CD8+ lymphocytes. | The results of the testing (correlation, linearity, reproducibility, cross-reactivity) as well as information from Leukocyte Typing Workshops indicate that the DAKO Anti-CD3/FITC plus Anti-CD8/RPE reagent performs as well as DAKO CD3/FITC in the detection and enumeration of CD3+ lymphocytes, and as well as DAKO CD8/RPE in the detection and enumeration of CD8+ lymphocytes using flow cytometry. |
2. Sample Size Used for the Test Set and Data Provenance:
- Sample Size: The document does not explicitly state a numerical sample size for the test set. It mentions "peripheral blood samples obtained from apparently healthy adults."
- Data Provenance: The data is from "peripheral blood samples obtained from apparently healthy adults." The country of origin is not specified, but the manufacturer is DAKO Corporation, located in Carpinteria, CA, USA, suggesting the study was likely conducted in the USA or a region where their products are distributed. The study appears to be prospective as it involves clinical evaluation comparing the new combined reagent to existing predicate devices.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of those Experts:
- This information is not provided in the summary. The ground truth appears to be established by the performance of the predicate devices themselves, which are already FDA-cleared. The comparison is against these established methods.
4. Adjudication Method:
- This information is not provided. The study design focuses on direct comparison of the new combined reagent with predicate single reagents.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done:
- No, an MRMC comparative effectiveness study was not done. This document describes a comparison between a new combined reagent and existing single reagents for flow cytometry, not a study evaluating human reader performance with or without AI assistance.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done:
- Yes, a standalone performance assessment was done. The device itself is a reagent (Mouse Anti-Human T-cell, CD3/FITC, UCHT1 + Mouse Anti-Human T-cell, CD8/RPE, DK25) used in flow cytometry. The study evaluates the performance of this reagent quantitatively (e.g., correlation coefficients for cell measurement, linearity), independent of human interpretation of the final flow cytometry data. The "performance characteristics have been established by clinical evaluation... compared to the individual single reagent predicate devices."
7. The Type of Ground Truth Used:
- The ground truth is based on the performance of previously FDA-cleared predicate devices. Specifically, DAKO Monoclonal Mouse Anti-Human T-cell, CD3/FITC, UCHT1 (K942797) and DAKO Monoclonal Mouse Anti-Human suppressor/cytotoxic T-cell, CD8/RPE, Clone DK25 (K944253). The implicit assumption is that these predicate devices provide an accurate measure of CD3+ and CD8+ cells.
8. The Sample Size for the Training Set:
- This information is not applicable as the device is a reagent, not a machine learning algorithm that requires a training set. The "training" in this context refers to the development and validation of the reagent itself.
9. How the Ground Truth for the Training Set Was Established:
- This information is not applicable for the same reason as point 8. If "training" refers to the development of the reagent, the establishment of its characteristics would have involved laboratory methods, chemical analysis, and potentially early comparison tests, but not in the sense of a machine learning algorithm's "training set ground truth." The document does mention "Leukocyte Typing Workshops" where antibody clones were clustered, indicating a consensus-based approach to defining these cell markers.
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MAR 28 8 1996
| 510(k )Summary K955909 | |
|---|---|
| Submitter: | DAKO Corporation6392 Via RealCarpinteria, CA 93013 |
| (805)566-6655 | |
| Contact: | Gretchen M. Murray, Ph.D., Regulatory Affairs Asst. Manager |
| Date Summary | |
| Prepared: | November 27, 1995 |
| Device Name: | Mouse Anti-Human T-cell, CD3/FITC, UCHT1 +Mouse Anti-Human T-cell, CD8/RPE, DK25 |
| DeviceClassification: | Class II according to 21 CFR 864.5220, on the basis that monoclonal antibodies |
| are accessories for automated differential cell counters. | |
| Panel: | This device classification is under the Hematology and Pathology devices panel,Division of Clinical Laboratory Devices. |
| Product Code: | GKZ |
| Predicate Device(s): | DAKO Monoclonal Mouse Anti-Human T-cell, CD3/FITC, UCHT1 (DAKO CodeNumber F0818, FDA K942797) andDAKO Monoclonal Mouse Anti-Human suppressor/cytotoxic T-cell, CD8/RPE, |
| Clone DK25 (DAKO Code Number R0806, FDA K944253) | |
| Device Description: | Purified mouse anti-human CD3, Clone UCHT1, conjugated with fluoresceinisothiocyanate, isomer 1 (FITC) + purified mouse anti-human CD8, Clone DK25,conjugated with R-phycoerythrin, present in 0.05M Tris-HCl buffer, pH 7.2, 15mM NaN3, 0.1M NaCl, stabilized with 1% carrier protein |
| Subpopulations of lymphocytes may be stained with fluorochrome-conjugatedantibody and evaluated in peripheral blood specimens when contaminating redblood cells (RBC's) are lysed prior to flow cytometric analysis. A subpopulationof WBC's are selected for assessment based upon cell morphology. | |
| Intended Use: | For In Vitro Diagnostic Use |
| Mouse Anti-Human T-cell, CD3/FITC, UCHT1 + Mouse Anti-Human T-cell,CD8/RPE, DK25 (DAKO Anti-CD3/FITC and Anti-CD8/RPE) has been developedfor use in flow cytometry for the analysis of CD3+ and CD8+ T-cells.Thisreagent allows simultaneous detection and quantification of CD3+CD8+ cells(CD8 positive T-lymphocytes) in normal and pathological conditions such asimmunodeficiency disorders. It is one component of the suggested monoclonalantibody (MAb) combination for routine immunophenotyping of lymphocytes inperipheral blood using flow cytometry. | |
| Comparison ofTechnologicalCharacteristics | Performance characteristics have been established by clinical evaluation ofcompared to the individual single reagent predicate devices that quantitativelymeasure CD3+ and CD8+ T-cells that have been previously cleared by FDA(DAKO CD3/FITC, Code No. F0818 and DAKO CD8/RPE, Code No. R0806).When flow cytometric tests of peripheral blood samples obtained fromapparently healthy adults were completed, correlation of Anti-CD3, UCHT1 with |
Nov. 27, 1995
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DAKO Anti-CD3/FITC and Anti-CD8/RPE approached a direct 1 : 1 comparison for measurement of CD3+ cells. Correlation of Anti-CD8, DK25 with DAKO Anti-CD3/FITC and Anti-CD8/RPE approached a direct 1 : 1 comparison for measurement of CD8 + cells. Data for the measurement of CD3 + T-cells by DAKO Anti-CD3/FITC and Anti-CD8/RPE reagent compared to DAKO CD3/FITC gave a correlation greater than 0.99 using the whole blood method for flow Data for the measurement of CD8+ T-cells by DAKO Anticytometry. CD3/FITC and Anti-CD8/RPE reagent compared to DAKO CD8/RPE gave a correlation greater than 0.98 usinq the whole blood method for flow cytometry.
The CD3 antibody clone. UCHT1, was clustered at the First Leukocyte Typing Workshop. Paris, France, 1982. The CD8 antibody clone, DK25, was clustered at the Third Leukocyte Typing Workshop, Oxford, England, 1986, under another clone designation.
Linearity testing of DAKO CD3/FITC using JM cells gave the following linear equation:
y = 0.02 + 0.98x; r = 0.999 Linearity testing of DAKO CD8/RPE using JM cells gave the following linear equation:
y = 0.06 + 1.01x; r = 0.999
In addition, reproducibility of DAKO reagents using replicates (from peripheral blood) run on two different flow cytometers was measured at three concentrations of each antigen. Cross-reactivity of Anti-CD3/FITC, plus Anti-CD8/RPE with peripheral blood cells (red blood cells, monocytes, granulocytes, lymphocytes, and platelets) was measured.
Results of the above testing as well as the information provided by the First and Third Leukocyte Typing Workshops indicate that the DAKO Anti-CD3/FITC plus Anti-CD8/RPE reagent performs as well as DAKO CD3/FITC in the detection and enumeration of CD3* lymphocytes while the DAKO Anti-CD3/FITC plus Anti-CD8/RPE reagent performs as well as DAKO CD8/RPE in the detection and enumeration of CD8 ' lymphocytes using flow cytometry. Safety of the DAKO Anti-CD3/FITC plus Anti-CD8/RPE reagent and its individual predicate devices is high as all reagents are used for in vitro testing.
Conclusions:
§ 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.”