(269 days)
The ClearLLab reagents are intended for in vitro diagnostic use as a panel for qualitative identification of cell populations by multiparameter immunophenotyping on an FC 500 flow cytometer. These reagents are used as an aid in the differential diagnosis of hematologically abnormal patients having, or suspected of having the following hematopoietic neoplasms; chronic leukemia, acute leukemia, non-Hodgkin lymphoma, myeloma, myelodysplastic syndrome (MDS), and/or myeloproliferative neoplasms (MPN). The reagents can be used with peripheral whole blood collected in EDTA (K2 and K3EDTA), Acid Citrate Dextrose or Heparin, bone marrow collected in K2EDTA, ACD, or Heparin, and lymph node specimens. Interpretation of the results should be confirmed by a pathologist or equivalent professional in conjunction with other clinical and laboratory findings.
These reagents provide multiparameter, qualitative results for the Cluster of Differentiation (CD) parameters listed below:
- ClearLLab T1: CD2, CD56, CD7, CD5, CD7, CD5, CD45
- ClearLLab T2: CD8, CD4, CD3, CD45
- ClearLLab B1: Kappa, Lambda, CD19, CD5, CD45
- ClearLLab B2: CD20, CD10, CD10, CD19, CD38, CD45
- ClearLLab M: CD7, CD13, CD34, CD33, CD45
The Clear LLab Reagents panel is used with the following reagents with their respective indicated uses. A description of the reagents provided is described below in Table 1.
| Item | Description | Use |
|---|---|---|
| The T-Cell Lineage panels | Comprising T1 CD2- | |
| FITC/CD56-PE/CD7- | ||
| ECD/CD5-PC5.5/CD45-PC7 | ||
| and T2 CD8-FITC/CD4- | ||
| PE/CD3-PC5.5/CD45-PC7 | ||
| Monoclonal Antibody Reagents. | For use to identify lymphocytes | |
| that contain the specific cell | ||
| surface markers associated with | ||
| the T-cell lineage. | ||
| The B-Cell Lineage panels | Comprising B1 Kappa- | For use to identify lymphocytes |
| FITC/Lambda-PE/CD19- | ||
| ECD/CD5-PC5.5/CD45-PC7 | ||
| and B2 CD20-FITC/CD10- | ||
| PE/CD19-ECD/CD38- | ||
| PC5.5/CD45-PC7 Monoclonal | ||
| and Polyclonal Antibody | ||
| Reagents. | that contain the specific cell | |
| surface markers associated with | ||
| the B-cell lineage. | ||
| The Myeloid Lineage panel | Comprising M CD7-FITC/ | |
| CD13-PE/CD34-ECD//CD33- | ||
| PC5.5/CD45-PC7 Monoclonal | ||
| Antibody Reagents. | For use to identify lymphocytes | |
| that contain the specific cell | ||
| surface markers associated with | ||
| the Myeloid lineage. | ||
| Accessory Reagents Required | ||
| Flow-Check Pro Fluorospheres | Flow-Check Pro Fluorospheres | |
| are a suspension of fluorescent | ||
| microspheres used for daily | ||
| verification of the optical | ||
| alignment and fluidics for | ||
| Forward Scatter (FS) and FL1- | ||
| FL4 fluorescence parameters. | Instrument Alignment Quality | |
| Control Reagent to provide | ||
| instrument alignment Quality | ||
| Control instructions. | ||
| Flow-Set Pro Fluorospheres | Flow-Set Pro Fluorospheres are | |
| a suspension of fluorescent | ||
| microspheres used as an aid in | ||
| standardizing forward scatter, | ||
| side scatter, and fluorescence | ||
| detectors on the FC 500 (FL1- | ||
| FL5). | Auto Setup Reagent for | |
| standardization of flow | ||
| cytometer light scatter and | ||
| fluorescence intensity | ||
| instrument settings to provide | ||
| application-specific instrument | ||
| target ranges for | ||
| standardization. | ||
| QuickComp 4 Kit | The QuickComp 4 Kit consists | |
| of four single-color fluorescent | ||
| reagents comprised of one | ||
| monoclonal antibody each. Each | ||
| antibody is labeled with one of | ||
| four fluorochromes, three are | ||
| utilized for this test: CD45- | ||
| FITC, CD45-PE, and CD45- | ||
| ECD. | The QuickComp 4 Kit is used to | |
| adjust color compensation | ||
| settings on a flow cytometer | ||
| equipped with AutoSetup | ||
| software, prior to multi-color | ||
| analysis with FITC, PE, and | ||
| ECD conjugated monoclonal | ||
| antibody reagents. | ||
| Color Compensation Reagents | Compensation reagents CD45- | |
| PC5.5 and CD45-PC7. | Used to adjust color | |
| compensation settings on a flow | ||
| cytometer with AutoSetup | ||
| software. | ||
| Note: Color Compensation | ||
| Reagents are required. It is | ||
| recommended to use the | ||
| reagents with normal whole | ||
| blood specimens to adjust color | ||
| compensation settings on a flow | ||
| cytometer, prior to multi-color | ||
| analysis. | ||
| IOTest 3 Fixative Solution | The IOTest 3 Fixative Solution, which has a formaldehyde base, specially developed for the fixing of leukocytes. | Leukocyte fixative solution used on all leukocytic preparations to enable leukocytic preparations to be stored for several hours without deterioration after staining with a fluorescent antibody. It is used to fix leukocytes following immunofluorescent staining with the fluorochrome-conjugated antibodies and lysis of the red blood cells. |
| Accessory Reagents Recommended but Not Provided | ||
| VersaLyse Lysing Solution | Red cell lysing reagent intended for the lysis of red blood cells in the preparation of biological samples for flow cytometry analysis. | VersaLyse is intended for the lysis of red blood cells in the preparation of biological samples for flow cytometry analysis. |
| Immutrol Control Cells | Process controls for flow cytometry. | Quality control material assayed for lymphocyte, granulocyte and monocyte specific antigens and single platform absolute counts. The light scatter, population distribution, fluorescence intensity, and antigen density mimic those of normal whole blood. |
| StemTrol Control cells | An antibody-to-antigen positive control for CD34 and CD45 staining in flow cytometry studies, consisting of a preserved cell line, BK010044, | Quality control material for CD34 and CD45. |
The ClearLLab Reagents are intended for in vitro diagnostic use as a panel for qualitative identification of cell populations by multiparameter immunophenotyping on an FC 500 flow cytometer. These reagents are used as an aid in the differential diagnosis of hematologically abnormal patients having, or suspected of having hematopoietic neoplasms.
1. A table of acceptance criteria and the reported device performance:
| Study/Metric | Acceptance Criteria (Target Outcome) | Reported Device Performance (Flow Expert #1) | Reported Device Performance (Flow Expert #2) |
|---|---|---|---|
| Overall Performance (All Specimen Types) | |||
| Diagnostic Agreement (Positive) | 80% ± 10% agreement with clinical diagnosis (70-90%) | Sensitivity: 82.4% (75.3%-87.8% CI) | Sensitivity: 85.9% (79.2%-90.7% CI) |
| Diagnostic Agreement (Negative) | 90% ± 10% agreement with clinical diagnosis (80-100%) | Specificity: 94.2% (88.9%-97.0% CI) | Specificity: 92.7% (87.1%-96.0% CI) |
| Whole Blood Specimen Performance | |||
| Diagnostic Agreement (Positive) | 90% ± 10% agreement with clinical diagnosis (80-100%) | Sensitivity: 89.0% (80.4%-94.1% CI) | Sensitivity: 92.7% (84.9%-96.6% CI) |
| Diagnostic Agreement (Negative) | 90% ± 10% agreement with clinical diagnosis (80-100%) | Specificity: 91.0% (82.6%-95.6% CI) | Specificity: 89.7% (81.0%-94.7% CI) |
| Qualitative Precision/Reproducibility | |||
| Repeatability (Presence/Absence of Abnormal Phenotype) | 100% agreement between expected and actual results | 100% agreement (all sites and anticoagulants) | 100% agreement (all sites and anticoagulants) |
| Operator-to-operator & Instrument-to-instrument Imprecision | 100% agreement for presence/absence of abnormal phenotype | 100% agreement (all operators and instruments) | 100% agreement (all operators and instruments) |
2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective):
- Test Set Sample Size: A total of 279 abnormal hematologic specimens were enrolled in the clinical performance study.
- 137 hematologically abnormal but no malignancy.
- 142 with hematolymphoid malignancy (23 Acute Leukemia, 36 Chronic Leukemia, 62 Lymphoma, 8 Plasma Cell Neoplasm, 13 Others: MDS/MPN).
- An additional analysis was performed on 160 whole blood specimens (78 hematologically abnormal with no malignancy and 82 with hematolymphoid malignancy) from this larger set.
- Data Provenance: The study was a multi-center, retrospective study conducted at four sites. While the document does not explicitly state the country of origin, the reference to "WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues" suggests an international context for clinical guidelines, but the specific locations of the four sites are not detailed.
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):
- Number of Experts: Two qualified flow experts evaluated the ClearLLab flow cytometry results.
- Qualifications of Experts: The document states "Two qualified flow experts evaluated the ClearLLab flow cytometry results independently of each other..." However, specific details on their years of experience or exact professional titles (e.g., "radiologist with 10 years of experience") are not provided. It does state that "The device labeling states that interpretation of specimens should be performed by a pathologist or equivalent professional who has the appropriate training." This implies the experts would hold such qualifications.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- Adjudication Method: The two flow experts evaluated the ClearLLab results independently of each other and were blinded to the final diagnosis. There is no mention of an adjudication method like 2+1 or 3+1 to resolve discrepancies between the two experts' interpretations of the ClearLLab device output. However, the expert interpretations of the ClearLLab results were then compared against the clinical outcome ("malignant" or "non-malignant") established by the clinical sites' final patient diagnosis.
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:
- This study did not appear to be an MRMC comparative effectiveness study in the context of human readers improving "with AI vs without AI assistance." The ClearLLab Reagents are a device for qualitative identification of cell populations using flow cytometry for aid in diagnosis. The study focused on the performance of these reagents (a diagnostic tool), interpreted by human experts, against clinical diagnosis. There is no mention of AI assistance in the interpretation of the device results or a comparison of human reader performance with and without such assistance.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:
- No, a standalone (algorithm only) performance was not done. The ClearLLab Reagents are a diagnostic tool that produces flow cytometry data. The interpretation of this data by human "flow experts" was an integral part of the clinical performance study. The device's output itself isn't a definitive diagnosis but an aid, requiring expert interpretation.
7. The type of ground truth used (expert concensus, pathology, outcomes data, etc):
- The ground truth used was the clinical outcome of "malignant" or "non-malignant" based on the clinical sites' final patient diagnosis. This diagnosis was established by a pathologist or equivalent professional in conjunction with other clinical and laboratory findings, adhering to WHO guidelines. This can be categorized as a form of outcomes data or established clinical diagnosis, rather than simply expert consensus on the device's output.
8. The sample size for the training set:
- The document focuses on the performance of the ClearLLab Reagents and associated interpretive processes. There is no mention of a "training set" in the context of an algorithm or AI model development, as this device submission is for reagents and their clinical performance when interpreted by human experts, not an AI diagnostic algorithm.
9. How the ground truth for the training set was established:
- As there is no mention of a "training set" for an algorithm or AI model in this document, the method for establishing its ground truth is not applicable or described.
§ 864.7010 Flow cytometric test system for hematopoietic neoplasms.
(a)
Identification. A flow cytometric test for hematopoietic neoplasms is a device that consists of reagents for immunophenotyping of human cells in relation to the level of expression, antigen density, and distribution of specific cellular markers. These reagents are used as an aid in the differential diagnosis or monitoring of hematologically abnormal patients having or suspected of having hematopoietic neoplasms. The results should be interpreted by a pathologist or equivalent professional in conjunction with other clinical and laboratory findings.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include the following information:
(i) The indications for use must indicate the clinical hematopoietic neoplasms for which the assay was designed and validated, for example, chronic leukemia or lymphoma.
(ii) A detailed device description including the following:
(A) A detailed description of all test components, all required reagents, and all instrumentation and equipment, including illustrations or photographs of nonstandard equipment or methods.
(B) Detailed documentation of the device software including, but not limited to, standalone software applications and hardware-based devices that incorporate software.
(C) A detailed description of methodology and assay procedure.
(D) A description of appropriate internal and external quality control materials that are recommended or provided. The description must identify those control elements that are incorporated into the testing procedure, if applicable.
(E) Detailed specifications for sample collection, processing, and storage.
(F) Detailed specification of the criteria for test results interpretation and reporting including pre-established templates.
(G) If applicable, based on the output of the results, a description of the specific number of events to collect, result outputs, and analytical sensitivity of the assay that will be reported.
(iii) Information that demonstrates the performance characteristics of the test, including:
(A) Device performance data from either a method comparison study comparing the specific lymphocyte cell markers to a predicate device or data collected through a clinical study demonstrating clinical validity using well-characterized clinical specimens. Samples must be representative of the intended use population of the device including hematologic neoplasms and the specific sample types for which the test is indicated for use.
(B) If applicable, device performance data from a clinical study demonstrating clinical validity for parameters not established in a predicate device of this generic type using well-characterized prospectively obtained clinical specimens including all hematologic neoplasms and the specific sample types for which the device is indicated for use.
(C) Device precision data using clinical samples to evaluate the within-lot, between-lot, within-run, between run, site-to-site and total variation using a minimum of three sites, of which at least two sites must be external sites. Results shall be reported as the standard deviation and percentage coefficient of variation for each level tested.
(D) Reproducibility data generated using a minimum of three lots of reagents to evaluate mean fluorescence intensity and variability of the recovery of the different markers and/or cell populations.
(E) Data from specimen and reagent carryover testing performed using well-established methods (
e.g., CLSI H26-A2).(F) Specimen and prepared sample stability data established for each specimen matrix in the anticoagulant combinations and storage/use conditions that will be indicated.
(G) A study testing anticoagulant equivalency in all claimed specimen type/anticoagulant combinations using clinical specimens that are representative of the intended use population of the device.
(H) Analytic sensitivity data using a dilution panel created from clinical samples.
(I) Analytical specificity data, including interference and cross-contamination.
(J) Device stability data, including real-time stability of reagents under various storage times and temperatures.
(K) For devices that include polyclonal antibodies, Fluorescence Minus One (FMO) studies to evaluate non-specific binding for all polyclonal antibodies. Each FMO tube is compared to reagent reference to demonstrate that no additional population appears when one marker is absent. Pre-specified acceptance criteria must be provided and followed.
(L) For devices indicated for use as a semi-quantitative test, linearity data using a dilution panel created from clinical samples.
(M) For devices indicated for use as a semi-quantitative test, clinically relevant analytical sensitivity data, including limit of blank, limit of detection, and limit of quantification.
(iv) Identification of risk mitigation elements used by the device, including a detailed description of all additional procedures, methods, and practices incorporated into the instructions for use that mitigate risks associated with testing the device.
(2) The 21 CFR 809.10 compliant labeling must include the following:
(i) The intended use statement in the 21 CFR 809.10(a)(2) and (b)(2) compliant labeling must include a statement that the results should be interpreted by a pathologist or equivalent professional in conjunction with other clinical and laboratory findings. The intended use statement must also include information on what the device detects and measures, whether the device is qualitative, semi-quantitative, and/or quantitative, the clinical indications for which the device is to be used, and the specific population(s) for which the device is intended.
(ii) A detailed description of the performance studies conducted to comply with paragraph (b)(1)(iii) of this section and a summary of the results.
(3) As part of the risk management activities performed under 21 CFR 820.30 design controls, product labeling and instruction manuals must include clear examples of all expected phenotypic patterns and gating strategies using well-defined clinical samples representative of both abnormal and normal cellular populations. These samples must be selected based upon the indications described in paragraph (b)(1)(i) of this section.