(60 days)
For In Vitro Diagnostic Use
Monoclonal Mouse Anti-Human T-cell, CD3/RPE-Cy5 conjugated, Clone UCHT1 (Code No.C7067) (Anti-CD3/RPE-Cy5) and Monoclonal Mouse Anti-Human T-cell, CD3/RPE conjugated, Clone UCHT1 (Code No. R0810) (Anti-CD3/RPE) have been developed for use in flow cytometry for the analysis of T-cells in peripheral blood. These reagents allow simultaneous detection and quantification of CD3-positive cells (T-cells) in normal and pathological conditions such as immunodeficiency disorders. Each reagent is one component of the suggested monoclonal antibody (MAb) combinations for routine immunophenotyping of lymphocytes in peripheral blood.
Immunophenotyping of lymphocytes is widely applied for diagnosis of immunodeficiencies. DAKO Anti-CD3/RPE-Cy5 is one of the reagents utilized when performing immunophenotyping of lymphocytes.
Monoclonal Mouse Anti-Human T-cell, CD3, Clone UCHT1, RPE conjugated, has been developed for use in flow cytometry for the analysis of T-cells. This reagent allows simultaneous detection and quantification of CD3-positive cells (T-cells) in normal and pathological conditions such as immunodeficiency disorders. It is one component of the suggested monoclonal antibody (MAb) combinations for routine immunophenotyping of lymphocytes in peripheral blood.
Immunophenotyping of lymphocytes is widely applied for diagnosis of immunodeficiencies. DAKO Anti-CD3/RPE is one of the reagents utilized when performing immunophenotyping of lymphocytes.
Monoclonal Mouse Anti-Human T-cell, CD3/RPE-Cy5 conjugated, Clone UCHT1 (Code No. C7067) and Monoclonal Mouse Anti-Human T-cell, CD3/RPE conjugated, Clone UCHT1 (Code No. R0810) are specific for T-lymphocyte cluster determinants as evaluated by the International Workshop on Human Leukocyte Differentiation Antigens. UCHT1 CD3-specific monoclonal antibody was designated B28 antibody at the Second Workshop (Reinherz, EL, Haynes, BF, Nadler, LM, Bernstein, ID, eds. Leukocyte Typing II, Vol. 2. New York-Berlin-Heidelberg-Tokyo: Springer Verlag, 1986.) Purified monoclonal mouse anti-human CD3 is produced in tissue culture, dialyzed and conjugated with either R-phycoerythrin (RPE) covalently coupled to cyanin 5 (Cy5) or R-phycoerythrin (RPE). One ml (1.0 ml) containing the conjugated antibody is supplied in 0.05M Tris-HCl buffer, pH 7.2, 15mM NaN3, 0.1M NaCl, stabilized with 1% stabilzing protein.
Monoclonal Mouse Anti-Human T-cell, CD3/RPE-Cy5-conjugated, C7067, and, Monoclonal Mouse Anti-Human T-cell, CD3/RPE, R0810 are specific for T-lymphocyte cluster determinants as evaluated by the International Workshop on Human Leukocyte Differentiation Antigens. UCHT1 CD3-specific monoclonal antibody was designated B28 antibody at the Second Workshop (Reinherz, EL, Haynes, BF, Nadler, LM, Bernstein, ID, eds. Leukocyte Typing II, Vol. 2. New York-Berlin-Heidelberg-Tokyo: Springer Verlag, 1986.) Purified monoclonal mouse anti-human CD3 is produced in tissue culture, dialyzed and conjugated with R-phycoerythrin (RPE) covalently coupled to cyanin 5 (Cy5), or conjugated with R-phycoerythrin (RPE). One ml (1.0 ml) containing the conjugated antibody is supplied in 0.05M Tris-HCl buffer, pH 7.2, 15mM NaNg, 0.1M NaCl, stabilized with 1% carrier protein.
Here's a summary of the acceptance criteria and the studies that prove the device meets these criteria, based on the provided text:
Device: DAKO® Mouse Anti-Human T-cell, CD3/RPE-Cy5, Clone UCHT1 (C7067) and DAKO® Mouse Anti-Human T-cell, CD3/RPE, clone UCHT1 (R0810)
Intended Use: For In Vitro Diagnostic Use in flow cytometry for the analysis of T-cells in peripheral blood, allowing simultaneous detection and quantification of CD3-positive cells (T-cells) in normal and pathological conditions. It is a component of suggested monoclonal antibody combinations for routine immunophenotyping of lymphocytes in peripheral blood.
1. Table of Acceptance Criteria and Reported Device Performance
| Acceptance Criteria | Device Performance (C7067) | Device Performance (R0810) |
|---|---|---|
| Linearity: Demonstrate a linear relationship between expected and recovered antigen concentrations. | y = -0.02 + 1.0x, R² = 0.999 (excellent linearity) | y = 2.84 + 1.0x, R² = 0.999 (excellent linearity) |
| Reproducibility: Consistent measurement of CD3+ cells across different antigen levels, instruments, and within a single day. (No explicit quantitative acceptance criteria for %CV or SD were stated, but low values are generally expected for good reproducibility.) | High Level: Mean % CD3+ 71.51 (SD 0.53, CV 0.01) on FACScan; 71.44 (SD 0.74, CV 0.01) on Profile II | High Level: Mean % CD3+ 72.07 (SD 1.11, CV 0.02) on FACScan; 71.53 (SD 0.74, CV 0.01) on Profile II |
| Medium Level: Mean % CD3+ 37.54 (SD 0.58, CV 0.02) on FACScan; 36.21 (SD 0.86, CV 0.02) on Profile II | Medium Level: Mean % CD3+ 38.50 (SD 0.87, CV 0.02) on FACScan; 36.01 (SD 0.49, CV 0.01) on Profile II | |
| Low Level: Mean % CD3+ 10.76 (SD 0.40, CV 0.04) on FACScan; 12.35 (SD 0.32, CV 0.03) on Profile II | Low Level: Mean % CD3+ 11.03 (SD 0.51, CV 0.05) on FACScan; 10.79 (SD 0.53, CV 0.05) on Profile II | |
| Specificity: Antibodies should specifically bind to lymphocytes (T-cells) with minimal binding to other cell types (RBCs, granulocytes, monocytes, platelets). | Average 71.90% positive Lymphocytes (range 60.3-78.6). Low binding to other cell types (e.g., 1.82% monocytes, 0.02% RBCs). | Average 69.92% positive Lymphocytes (range 58.4-75.9). Low binding to other cell types (e.g., 8.52% monocytes, 0.08% RBCs). |
| Predicate Correlation: High correlation and comparable performance to the predicate device (DAKO Anti-CD3/FITC, F0818) in peripheral blood samples from healthy individuals and patients with illnesses. (Implicitly, a high R² value close to 1 and a slope close to 1 with a small intercept are desired). | Healthy (n=150): Y = 6.22 + 0.92X, R² = 0.9208 | |
| Total (n=177): Y = 2.43 + 0.98X, R² = 0.9750 | Healthy (n=150): Y = 9.85 + 0.87X, R² = 0.8040 | |
| Total (n=176): Y = 1.20 + 0.99X, R² = 0.9870 |
2. Sample Sizes Used for Test Set and Data Provenance
- Linearity Test Set: 5 serial dilutions of a cell line expressing the antigen (JM cells) diluted with a cell line without antigenic sites (Raji cells). Data provenance not specified (likely in-house lab, prospective).
- Reproducibility Test Set: 10 replicates from peripheral blood of "three donors" at three concentrations (high, medium, low). Data provenance not specified (likely in-house lab, prospective).
- Specificity Test Set: 5 apparently healthy adult donors of various races. Data provenance: DAKO Corporation (in-house lab, prospective).
- Predicate Correlation Test Set:
- Healthy Individuals: 153 normal, apparently healthy individuals.
- Patient Samples: An unspecified number of samples from "patients with illnesses" were added to the healthy individual data.
- Total for C7067: 177 samples (150 healthy + 27 patient samples).
- Total for R0810: 176 samples (150 healthy + 26 patient samples).
- Data Provenance: Three geographically separate laboratories (likely prospective, multi-site study).
3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications
The studies described are for the performance characteristics of an in vitro diagnostic reagent (antibody). The "ground truth" in this context is based on the inherent molecular specificity of the antibody and its performance in flow cytometry, rather than expert interpretation of complex images or clinical cases.
- No "experts" in the sense of clinicians or radiologists establishing a diagnostic ground truth were used.
- The ground truth for antigen expression was established by using known cell lines (JM cells for expression, Raji cells for no expression) in the linearity study, and by the established methodology of flow cytometry for reproducibility and specificity.
- The predicate correlation study compares the new reagents to a previously validated predicate, implicitly using the predicate's results as a reference.
4. Adjudication Method for the Test Set
Not applicable. This is not a study requiring adjudication of interpretations (e.g., of images or clinical reports) by human readers. The measurements are quantitative outputs from flow cytometry.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done
No, an MRMC comparative effectiveness study was not done. This device is a reagent for flow cytometry, not an algorithm or system that directly aids human readers in interpretation or diagnosis. The study assessed the reagent's performance against established laboratory standards and a predicate device.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the studies conducted (linearity, reproducibility, specificity, predicate correlation) assess the standalone performance of the reagents themselves in a laboratory setting when used in flow cytometry. While flow cytometry involves human operation, the performance metrics reported are for the reagent's ability to bind to target cells and be quantified, not for human interpretation aided by an algorithm.
7. The Type of Ground Truth Used
- Linearity: Known concentrations of antigen-expressing cells (JM cells) mixed with non-expressing cells (Raji cells).
- Reproducibility: Target population of CD3+ lymphocytes within peripheral blood samples, as measured by flow cytometry. The reference values are the means of the repeated measurements.
- Specificity: Known cell populations (RBCs, granulocytes, monocytes, lymphocytes, platelets) obtained from healthy donors. The expected "ground truth" is that the antibody should primarily bind to lymphocytes (T-cells).
- Predicate Correlation: The results obtained using the legally marketed predicate device (DAKO Mouse anti-human T-cell, CD3/FITC, Clone UCHT1 Code No. F0818) serve as the reference standard.
8. The Sample Size for the Training Set
The provided text does not explicitly mention a "training set" in the context of machine learning or algorithm development. The device is a diagnostic reagent (antibody) for flow cytometry, and its development typically involves laboratory characterization rather than iterative training on large datasets like an AI algorithm. If "training" refers to the development and initial characterization of the antibody itself:
- The antibody (UCHT1) was designated at the "Second Workshop" (International Workshop on Human Leukocyte Differentiation Antigens), implying a historical development and characterization process that predates this submission.
- The production of purified monoclonal mouse anti-human CD3 is described, but the "training set" size for its initial development or characterization is not specified in this document.
9. How the Ground Truth for the Training Set Was Established
As noted above, a "training set" in the machine learning sense is not directly applicable here. The "ground truth" for the antibody's properties (specificity to CD3, binding characteristics) would have been established through:
- Immunological assays: Early characterization during antibody development to confirm its binding to the CD3 antigen on T-cells.
- International Workshops: The antibody's designation (B28 antibody at the Second Workshop) indicates that its specificity and reactivity were validated and recognized by the scientific community, likely through extensive testing against various cell types and patient samples, establishing its utility as a marker for CD3. This historical validation forms the basis of its "ground truth" for specificity.
§ 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.”