(203 days)
No
The summary describes a flow cytometry-based assay using monoclonal antibodies to identify and enumerate fetal red blood cells. The analysis involves standard flow cytometry methods and statistical analysis of the results (e.g., calculating percentages, standard deviations, correlation coefficients). There is no mention of AI, ML, or any advanced computational techniques for data interpretation or classification beyond standard flow cytometry gating and analysis.
No
The device is an in vitro diagnostic (IVD) test intended for the identification and enumeration of fetal red blood cells, which aids in detecting incompatible fetal-maternal hemorrhage and determining the need for immunoprophylaxis. It does not provide any direct therapeutic benefit to the patient.
Yes
The device "is intended for the identification followed by enumeration of fetal red blood cells," and "may be used as an aid in detecting incompatible fetal-maternal hemorrhage and determining the need for immunoprophylaxis with Rh immune globulin," which describes a diagnostic purpose.
No
The device description details a laboratory test involving physical blood samples, chemical reagents (monoclonal antibodies), and analysis using a flow cytometer, which is a hardware instrument. This is not a software-only device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states it is "intended for the identification followed by enumeration of fetal red blood cells" and "may be used as an aid in detecting incompatible fetal-maternal hemorrhage and determining the need for immunoprophylaxis with Rh immune globulin." This clearly describes a diagnostic purpose performed in vitro (on a blood sample).
- Device Description: The description details the process of analyzing an anticoagulated peripheral blood sample using monoclonal antibodies and flow cytometry. This is a typical in vitro diagnostic procedure.
- Predicate Device: The presence of a predicate device with the name "Sure-Tech Fetal Hemoglobin Test for in vitro diagnostic use" further confirms that this type of device falls under the IVD category.
N/A
Intended Use / Indications for Use
The Caltag Fetal Hemoglobin Test, containing monoclonal antibodies to fetal hemoglobin (hemoglobin F) conjugated with either FITC, R-PE or TRI-COLOR®, is intended for the identification followed by enumeration of fetal red blood cells in the maternal circulation. Fetal cells are identified by the presence of fetal hemoglobin by a flow cytometric method. The presence of fetal cells in the maternal circulation, resulting from fetal-maternal hemorrhage, may be attributed to a variety of causes, including fetal trauma, various obstetrical emergencies and placental trauma. Hemorrhage of Rh+ fetal blood into Rh- maternal blood may result in the formation of sensitizing Rh antibodies in the mother. This Rh immunization may be prevented by the administration to the mother of Rh immune globulin (RhIg) soon after delivery. The Caltag Fetal Hemoglobin Test may be used as an aid in identifying fetal-maternal hemorrhage and determining the need for immunoprophylaxis with immune globulin.
Product codes
GHQ
Device Description
The Caltag Fetal Hemoglobin Test consists of monoclonal antibodies to fetal hemoglobin (hemoglobin F) conjugated with either FITC, R-PE or TRI-COLOR®. An anticoagulated peripheral blood sample is drawn, erythrocyte count is determined and adjusted, followed by brief fixation in gluteraldehyde. Fixed and washed cells are permeabilized with a detergent to enable monoclonal antibodies to penetrate cellular membranes. Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR monoclonal antibodies bind to fetal hemoglobin in fetal red cells. To identify cells containing fetal hemoglobin, fixed and permeabilized cells are incubated with the monoclonal antibody, and washed to remove unbound antibody. Antibody stained cells are subsequently analyzed by flow cytometric methods. Positive and negative control samples must be used to establish reagent performance and differentiate positive fluorescence from unstained normal red blood cells, leukocytes and cellular debris. Recommended positive control samples consist of 1% and 5% fetal erythrocyte-containing placental cord blood in normal adult blood. The recommended negative control sample consists of 1% anticoagulated sample from a normal male or non-pregnant adult female.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Maternal Circulation (specifically peripheral blood sample)
Indicated Patient Age Range
Not Found (The test is for maternal circulation, implying adult women of childbearing age, but no specific age range is mentioned for the patients being tested. Expected value data was collected from adult female donors aged 19-50.)
Intended User / Care Setting
Not Found (Implied to be a clinical laboratory setting based on the nature of the test and the Clinical Laboratory Improvement Amendments (CLIA-88) reference.)
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Non-Clinical Tests:
- Expected Value Data:
- Study Type: Determination of normal range. Blood samples analyzed for expected values of FITC, R-PE, and TRI-COLOR monoclonal antibodies to HbF.
- Sample Size: 161 adult female normal donors.
- Data Source: Blood samples collected from donors aged 19-50 (mean/median 33) from three independent laboratories in geographically diverse areas (Northern, South-central, Western regions of US), with varying ethnic origins (Caucasians, Blacks, Orientals, Hispanics).
- Key Results: 95% Reference Intervals (Normal Range) for percent positive: FITC (0.00-0.15), R-PE (0.00-0.14), TRI-COLOR (0.00-0.14).
- Specificity Data:
- Study Type: Evaluation of staining of blood elements other than fetal erythrocytes.
- Data Source: Blood samples from healthy normal donors of Caucasian, Black, Hispanic, and Oriental ethnic origins. Stained with Caltag HbF FITC, HbF R-PE, and HbF TRI-COLOR monoclonal antibodies. Cells from lymphocyte, monocyte, granulocyte regions, red blood cells, and platelets were analyzed.
- Key Results: No staining of any non-fetal erythrocyte blood elements by Caltag HbF monoclonal antibodies.
- Reproducibility Data (Intra-lab):
- Study Type: Assessed intra-laboratory reproducibility at high, medium, and low ranges for each antibody.
- Sample Size: 6 replicated determinations for each antibody (FITC, R-PE, TRI-COLOR) in each of three ranges (high, medium, low), totaling 18 determinations per antibody. Samples consisted of varying mixtures of placental cord blood in normal adult blood.
- Data Source: Performed in three independent laboratories, each obtaining, staining, and analyzing separate blood samples.
- Key Results (Representative): Low %CV values across all procedures and levels (e.g., HbF FITC: high 2.42% CV, mid 4.30% CV, low 2.26% CV).
- Reproducibility Data (Inter-lab):
- Study Type: Assessed inter-laboratory reproducibility at high, medium, and low ranges for each antibody.
- Sample Size: Similar to intra-lab: 6 replicated determinations for each antibody and range. Samples consisted of varying mixtures of placental cord blood in normal adult blood.
- Data Source: Performed in three independent laboratories, all laboratories stained and analyzed the same samples prepared by one participating lab.
- Key Results: Low %CV values indicating good inter-lab reproducibility across all procedures and levels for each site.
- Linearity of Measurement:
- Study Type: Confirmed linearity of measurement for samples with 0.0-5.0% cord blood cells.
- Data Source: Samples consisting of mixtures of cord blood cells in normal adult blood.
- Key Results: High r-squared values (HbF FITC: 99.97, HbF R-PE: 99.96, HbF Tri-Color: 99.98) and slopes near 1.0, and Y intercepts near 0.
- Saturating Conditions Study:
- Study Type: Determined the ability of the test to detect all fetal cells in 100% cord blood samples.
- Sample Size: 5 different cord blood samples.
- Data Source: Stained and analyzed in a single laboratory.
- Key Results: High percentage of fetal cells detected (HbF FITC: 95.66%, HbF R-PE: 96.70%, HbF TRI-COLOR: 95.60%), indicating sufficient concentration and potency of antibodies.
Clinical Tests:
- Correlation Data:
- Study Type: Correlation against Kleihauer-Betke (KB) microscopic staining method.
- Data Source: Conducted in 3 independent laboratories.
- Study Site #1:
- Sample Size: Patient samples (n=30) and prepared samples (n=50) analyzed with Caltag HbF monoclonal antibodies (FITC, R-PE, TC) and KB test.
- Patient Samples Data: Patient samples from women with clinical indications of fetal-maternal hemorrhage.
- Prepared Samples Data: Mixtures of fetal cord blood in normal adult blood (0-10% fetal cells), analyzed on Becton Dickinson FACscan and Coulter EPICS-XL flow cytometers.
- Key Results (Prepared Samples on FACscan): High r-squared values for correlations between Caltag tests and KB test (97.95-98.16) and between different Caltag antibodies (99.59-99.70). Mean % Positive values similar between Caltag and KB.
- Key Results (Prepared Samples on EPICS-XL): High r-squared values for correlations between Caltag tests and KB test (97.96-98.48) and between different Caltag antibodies (99.54-99.67). Mean % Positive values similar between Caltag and KB.
- Key Results (Patient Samples on FACscan): High r-squared values for correlations between Caltag tests and KB test (96.80-97.50). Mean % Positive values similar between Caltag and KB.
- Study Site #2:
- Sample Size: Patient samples (n=38) and prepared samples (n=15) for HbF FITC monoclonal antibody and KB test.
- Data Range: 0-3.0%.
- Key Results (Patient Samples on EPICS-XL): r2 value of 98.34, mean % positive of 0.22 (HbF FITC) vs 0.20 (KB Method).
- Key Results (Prepared Samples on EPICS-XL): r2 value of 85.50, mean % positive of 1.52 (HbF FITC) vs 1.61 (KB Method).
- Study Site #3:
- Sample Size: Patient samples (n=13) and prepared samples (n=15) for HbF R-PE monoclonal antibody and KB test.
- Data Range: 0-3.0%, analyzed on EPICS-XL flow cytometer.
- Key Results (Patient Samples on EPICS-XL): r2 value of 64.00, mean % positive of 0.08 (HbF R-PE) vs 0.11 (KB Method).
- Key Results (Prepared Samples on EPICS-XL): r2 value of 84.01, mean % positive of 1.38 (HbF R-PE) vs 1.40 (KB Method).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Specificity: Specificity study indicated no staining of any blood elements other than fetal erythrocytes by the Caltag HbF monoclonal antibodies.
- Reproducibility (Intra-lab and Inter-lab): Reported as %CV (coefficient of variation). For intra-lab, %CV ranged from 2.26 to 4.75 for FITC, R-PE, and TC across high, mid, and low levels. For inter-lab, %CV ranged from 1.37 to 7.40 across sites, antibodies, and levels.
- Linearity (r2 value): HbF FITC: 99.97, HbF R-PE: 99.96, HbF Tri-Color: 99.98.
- Correlation (r2 value):
- Between Caltag (FITC, R-PE, TC) and KB on prepared samples (FACscan): 97.95 - 98.16.
- Between Caltag (FITC, R-PE, TC) and KB on prepared samples (EPICS-XL): 97.96 - 98.48.
- Between Caltag (FITC, R-PE, TC) and KB on patient samples (FACscan) in Site 1: 96.80 - 97.50.
- Between Caltag (FITC) and KB on patient samples (EPICS-XL) in Site 2: 98.34.
- Between Caltag (FITC) and KB on prepared samples (EPICS-XL) in Site 2: 85.50.
- Between Caltag (R-PE) and KB on patient samples (EPICS-XL) in Site 3: 64.00.
- Between Caltag (R-PE) and KB on prepared samples (EPICS-XL) in Site 3: 84.01.
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 864.7455 Fetal hemoglobin assay.
(a)
Identification. A fetal hemoglobin assay is a device that is used to determine the presence and distribution of fetal hemoglobin (hemoglobin F) in red cells or to measure the amount of fetal hemoglobin present. The assay may be used to detect fetal red cells in the maternal circulation or to detect the elevated levels of fetal hemoglobin exhibited in cases of hemoglobin abnormalities such as thalassemia (a hereditary hemolytic anemia characterized by a decreased synthesis of one or more types of hemoglobin polypeptide chains). The hemoglobin determination may be made by methods such as electrophoresis, alkali denaturation, column chromatography, or radial immunodiffusion.(b)
Classification. Class II (special controls). A fetal hemoglobin stain intended for use with a fetal hemoglobin assay is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 864.9.
0
SEP 1 7 1999
510(K) SUMMARY SUMMARY OF SAFETY AND EFFECTIVENESS DATA
BORATORIE:
Caltag Fetal Hemoglobin Test for the Detection of Fetal Red Blood Cells in the Maternal Circulation (Fetal-Maternal Hemorrhage)
NAME AND LOCATION OF THE MANUFACTURER: Caltag Laboratories, Inc. 1849 Old Bayshore Highway Suite 200 Burlingame, CA 94010 (800) 874-4007
NAME OF CONTACT PERSON: Robert C. Johnson Executive Vice President Caltag Laboratories, Inc.
DATE OF PREPARATION OF THE SUMMARY: July 09, 1999
TRADE NAME OF THE DEVICE:
Caltag Fetal Hemoglobin Test for the Detection of Fetal Red Blood Cells in the Maternal Circulation (Fetal-Maternal Hemorrhage) ・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・
İ
- 1
COMMON NAME:
Caltag Fetal Hemoglobin Test
CLASSIFICATION NAME:
Fetal Hemoglobin Assay; 21CFR 864.7455 Product Code: GHQ
LEGALLY MARKETED (PREDICATE) DEVICE TO WHICH THE MANUFACTURER IS CLAIMING SUBSTANTIAL EQUIVALENCE:
Caltag Fetal Hemoglobin Test is substantially equivalent to Sure-Tech Fetal Hemoglobin Test for in vitro diagnostic use (K892241), manufactured by Sure-Tech Diagnostic Associates, Inc., 11012 Un Valle, Suite D, St. Louis, Mo. 63123,
1
An anticoagulated peripheral blood sample is drawn from an appropriate donor. The erythrocyte count is determined and adjusted, followed by brief fixation of the cells in gluteraldehyde.
Fixed and washed cells are permeabilized with a detergent in a manner that is frequently used to enable macromolecules such as monoclonal antibodies to penetrate cellular membranes. Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR monoclonal antibodies bind to fetal hemoglobin in fetal red cells. To identify cells containing fetal hemoglobin, fixed and permeabilized cells are incubated with the monoclonal antibody, and washed to remove unbound antibody. Antibody stained cells are subsequently analyzed by flow cytometric methods.
Positive and negative control samples must be used with sample analysis, to establish that all reagents are performing in a consistent manner and that the positive fluorescence attributed to antibody-stained fetal red cells is differentiated from unstained normal red blood cells, leukocytes and any cellular debris. If cord blood is not available for the performance of positive controls, the assay cannot be performed reliably. The recommended positive control samples consist of both 1% and 5% fetal erythrocyte-containing placental cord blood in normal adult blood. The recommended negative control sample consists of 1% anticoagulated sample from a normal male or non-pregnant adult female.
INTENDED USE OF THE DEVICE:
The Caltag Fetal Hemoglobin Test, containing monoclonal antibodies to fetal hemoglobin (hemoglobin F) conjugated with either FITC, R-PE or TRI-COLOR®, is intended for the identification followed by enumeration of fetal red blood cells. Fetal cells are identified by the presence of fetal hemoglobin by a flow cytometric method. The presence of fetal cells in the maternal circulation, resulting from fetalmaternal hemorrhage, may be attributed to a variety of causes, including fetal trauma, various obstetrical emergencies and placental trauma. Hemorrhage of Rh+ fetal blood into Rh- maternal blood may result in the formation of sensitizing Rh antibodies in the mother. This Rh immunization may be prevented by the ---administration to the mother of Rh immune globulin (RhIg) soon after delivery. The Caltag Fetal Hemoglobin Test may be used as an aid in identifying fetal-maternal hemorrhage and determining the need for immunoprophylaxis with immune globulin.
SUMMARY OF TECHNICAL CHARACTERISTICS OF THE DEVICE COMPARED TO THE PREDICATE DEVICE:
| No. | Item | Caltag Test | Predicate Test | Comparison |
|---|---|---|---|---|
| 1. | Intended Use | Enumeration of | ||
| Fetal cells | Enumeration of | |||
| Fetal cells | Substantially | |||
| equivalent | ||||
| 2. | Specificity | Fetal hemoglobin | Fetal hemoglobin | Substantially |
| equivalent | ||||
| 3. | Target cell | Fetal erythrocyte | Fetal erythrocyte | Substantially |
| equivalent |
Comparisons of Caltag and Predicate Fetal Hemoglobin Tests
2
| Continued.. | ||||
|---|---|---|---|---|
| 4. | Method of | |||
| Detection | Flow Cytometry | Light Microscopy | Different | |
| methods used | ||||
| 5. | Chemical form | Monoclonal | ||
| antibody | Pink staining with | |||
| erythrocin | Substantially | |||
| equivalent | ||||
| 6. | Device | |||
| Configuration | Diagnostic Kit | |||
| with Reagents | Diagnostic Kit | |||
| with Reagents | Substantially | |||
| equivalent | ||||
| 7. | Sample prep. | |||
| methods | Fixed, Permeabilized | |||
| Whole Blood | Whole Blood | Substantially | ||
| equivalent | ||||
| 8. | Correlation of | |||
| Methods (FACscan & EPICS-XL Site) | ||||
| Prepared Samples (n=50) | Substantially | |||
| Mean % Positive | FITC 4.54 | KB 4.51 | equivalent | |
| r squared value | 97.95 | |||
| Mean % Positive | R-PE 4.50 | KB 4.41 | ||
| r squared value | 98.16 | |||
| Mean % Positive | TC 4.49 | KB 4.41 | ||
| r squared value | 97.98 | |||
| Patient Samples (n=30) | Substantially | |||
| Mean % Positive | FITC 0.51 | KB 0.51 | equivalent | |
| r squared value | 96.80 | |||
| Mean % Positive | R-PE 0.50 | KB 0.51 | ||
| r squared value | 96.84 | |||
| Mean % Positive | TC 0.47 | KB 0.51 | ||
| r squared value | 97.50 |
NON-CLINICAL TESTS SUPPORTING A DETERMINATION OF SUBSTANTIAL EQUIVALENCE:
Expected Value Data:
Blood samples were collected from a total of 161 adult female normal donors in an age range of 19-50, with a mean and median age of 33. These consisted of ------approximately 50 female normal donors in each of three independent laboratories. In each laboratory, normal donors were distributed approximately equally among the three age ranges of 19-29, 30-39 and 40-50. A single sample from each donor was analyzed for the determination of expected values of the Caltag FITC. R-PE and TRI-COLOR® monoclonal antibodies to HbF.
The normal donor population consisted of members of differing ethnic origins, including adult Caucasians, Blacks, Orientals and Hispanics. The three independent laboratories were located in geographically diverse areas within the United States, including the Northern, South-central and Western regions.
Normal ranges are expressed as the 95% reference intervals (17), consisting of the mid-95% of values for all donors when rank ordered from lowest to highest percent of fetal cells detected, as indicated on the following table;
3
EXPECTED VALUES IN ALL ADULT FEMALE NORMAL DONORS
| procedure | mean
% positive | S.D. | 95% Reference
Interval
(Normal Range) | n |
|-----------|--------------------|------|---------------------------------------------|-----|
| FITC | 0.03 | 0.04 | 0.00--0.15 | 153 |
| R-PE | 0.04 | 0.05 | 0.00--0.14 | 153 |
| TRI-COLOR | 0.03 | 0.04 | 0.00--0.14 | 153 |
Specificity Data:
This study was conducted to evaluate any staining of blood elements other than fetal erythrocytes by the Caltag HbF monoclonal antibodies. Blood samples were obtained from healthy normal donors of Caucasian, Black, Hispanic and Oriental ethnic origins.
Samples of each donor were stained with Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR monoclonal antibodies. Cells contained in the lymphocyte, monocyte and granulocyte regions were selected for analysis. Separate sample were prepared for analysis of red blood cells and platelets and were stained with each of the Caltag monoclonal antibodies.
The specificity study indicated no staining of any of the indicated blood elements by the Caltag HbF monoclonal antibodies in samples from normal donors.
Reproducibility Data: intra-lab
Intra-lab reproducibility for the Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR (TC) conjugated monoclonal antibodies was determined by performing 6 replicated determinations for each antibody in each of three ranges: high, medium and low. Thus, a total of 18 determinations were performed for each HbF antibody. In this manner, reproducibility was demonstrated throughout the entire measurement range for the Caltag Fetal Hemoglobin Test.
The 6 determinations for each range were performed by the staining, processing and analysis of 6 separate samples consisting of varying mixtures of placental cord blood in normal adult blood. Fetal cells were selected for the analysis of percent cells stained in each of the three ranges.
The study was performed in each of three independent laboratories, in the manner that each laboratory obtained, stained and analyzed separate blood samples.
The following data are representative:
| procedure | Level | mean
% positive | S.D. | % CV | n |
|-----------|-------|--------------------|------|------|---|
| HbF FITC | high | 9.51 | 0.22 | 2.42 | 6 |
| | mid | 5.10 | 0.22 | 4.30 | 6 |
| | low | 2.19 | 0.05 | 2.26 | 6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF R-PE | high | 9.61 | 0.40 | 4.25 | 6 |
| | mid | 5.10 | 0.14 | 2.66 | 6 |
| | low | 2.14 | 0.10 | 4.75 | 6 |
4
| procedure | Level | mean
% positive | S.D. | % CV | n |
|-----------|-------|--------------------|------|------|---|
| HbF TC | high | 9.64 | 0.37 | 3.84 | 6 |
| | mid | 5.13 | 0.21 | 4.16 | 6 |
| | low | 2.23 | 0.10 | 4.58 | 6 |
Reproducibility Data, inter-lab
Inter-lab reproducibility for the Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR (TC) conjugated monoclonal antibodies was determined by performing 6 replicated determinations for each antibody in each of three ranges; high, medium and low. In this manner, reproducibility was demonstrated throughout the entire measurement range for the Caltag Fetal Hemoglobin Test.
The 6 determinations for each range were performed by the staining, processing and analysis of 6 separate samples consisting of varying mixtures of placental cord blood in normal adult blood. Fetal cells were selected for the analysis of percent cells stained in each of the three ranges.
The study was performed in each of three laboratories. All laboratories stained and analyzed the same samples.
Unstained and unfixed samples containing mixtures of cord blood in normal adult blood representing the appropriate ranges were prepared by one of the participating laboratories for staining and analysis by each of the participating laboratories. The following data were obtained:
| SITE 1
procedure | Level | mean
% positive | S.D. | % CV | n |
|---------------------|-------|--------------------|------|------|---|
| HbF FITC | high | 6.34 | 0.09 | 1.48 | 6 |
| | mid | 4.05 | 0.08 | 2.09 | 6 |
| | low | 1.48 | 0.04 | 2.91 | 6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF R-PE | high | 7.90 | 0.29 | 3.64 | 6 |
| | mid | 5.35 | 0.30 | 5.60 | 6 |
| | low | 2.84 | 0.21 | 7.34 | 6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF TC | high | 7.43 | 0.10 | 1.37 | 6 |
| | mid | 4.37 | 0.11 | 2.57 | 6 |
| | low | 1.60 | 0.07 | 4.55 | 6 |
| SITE 2
procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF FITC | high | 8.84 | 0.21 | 2.32 | 6 |
| | mid | 5.69 | 0.13 | 2.36 | 6 |
| | low | 2.44 | 0.08 | 3.28 | 6 |
-5-
5
| procedure | Level | mean
% positive | S.D. | % CV | n |
|---------------------|--------------------|----------------------|----------------------|----------------------|-------------|
| HbF R-PE | high
mid
low | 8.81
5.70
2.36 | 0.22
0.14
0.04 | 2.51
2.45
1.89 | 6
6
6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF TC | high
mid
low | 8.82
5.68
2.41 | 0.12
0.15
0.05 | 1.39
2.69
1.90 | 6
6
6 |
| SITE 3
procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF FITC | high
mid
low | 8.95
5.85
2.57 | 0.36
0.23
0.14 | 4.04
3.86
5.32 | 6
6
6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF R-PE | high
mid
low | 8.43
5.60
2.38 | 0.12
0.41
0.07 | 1.44
7.40
3.16 | 6
6
6 |
| procedure | Level | mean
% positive | S.D. | % CV | n |
| HbF TC | high
mid
low | 8.70
5.65
2.41 | 0.46
0.18
0.17 | 5.24
3.12
7.13 | 6
6
6 |
Linearity of measurement was determined for samples consisting of mixtures of cord blood cells in normal adult blood in the range of 0.0-5.0% cord blood cells. This range was selected to represent the entire range of values for the percent of fetal cells that are likely to be experienced in fetal-maternal hemorrhage. The linear regression method was used to plot the known expected values versus the observed values for the percent of fetal cells determined by the flow cytometric method for the Caltag FITC, R-PE and TRI-COLOR® monoclonal antibodies to HbF:
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
LINEARITY OF MEASUREMENT
| procedure | r2 value | slope | Y intercept | n |
|---|---|---|---|---|
| HbF FITC | 99.97 | 1.01 | -0.04 | 10 |
| HbF R-PE | 99.96 | 1.02 | -0.03 | 10 |
| HbF Tri-Color | 99.98 | 1.02 | -0.02 | 10 |
6
An additional study was conducted to identify saturating conditions for each of the Caltag monoclonal antibodies, consisting of preparations of cord blood alone (100% cord blood). Five different cord blood samples were stained and analyzed in a single laboratory, to determine the ability of the Caltag Fetal Hemoglobin Test to detect all fetal cells present, and to assure that the concentration and potency of the HbF monoclonal antibodies is sufficient to detect all fetal cells in a test sample. Therefore, samples employed in this study contained a greater than tenfold higher content of fetal cells than would be expected to occur in clinical specimens.
Summary of values obtained for the percent of fetal cells detected by the Caltag HbF FITC, HbF R-PE and HbF TRI-COLOR antibodies in 100% cord blood samples:
| procedure | mean
% positive | S.D. | % CV | n |
|---------------|--------------------|------|------|---|
| HbF FITC | 95.66 | 3.88 | 4.05 | 5 |
| HbF R-PE | 96.70 | 2.70 | 2.80 | 5 |
| HbF TRI-COLOR | 95.60 | 2.58 | 2.70 | 5 |
CLINICAL TESTS SUPPORTING A DETERMINATION OF SUBSTANTIAL EQUIVALENCE
Correlation Data:
The Correlation study was conducted in 3 independent laboratories. Samples were analyzed with the Caltag Fetal Hemoglobin Test and a commercially available test based on the Kleihauer-Betke (KB) microscopic staining method for the detection of fetal hemoglobin in fetal cells (4) in each site. In all studies, confidence intervals are expressed at the 95% limit.
In study site #1, the percent of fetal cells detected by each of the HbF monoclonal antibodies to fetal hemoglobin was correlated with the percent of fetal cells detected by the KB test on patient samples (n=30) and prepared samples (n=50). Patient samples were obtained from women having clinical indications that were consistent with fetal-maternal hemorrhage and prepared samples consisted of mixtures of fetal cord blood in normal adult blood prepared in differing cell ratios in the range of 0-10% fetal cells. This range included up to the equivalent of 500. ml of fetal blood in maternal blood and greatly exceeded the 150--300 ml of fetal blood encountered in the most severe cases of fetal-maternal hemorrhage (3).
In study sites #2, the percent of fetal cells detected by the HbF FITC monoclonal antibody to fetal hemoglobin was correlated with the percent of fetal cells detected by the KB test in patient samples (n = 38) and prepared samples (n = 15) in the range of 0-3.0%. The Caltag test was propared Sumploo (n ----------------------------------------------------------------------------------------------------------------------------------------------------------
In study site #3, the percent of fetal cells detected by the HbF R-PE monoclonal to fetal hemoglobin was correlated with the percent of fetal cells detected by the KB test in patient samples (n = 13) and prepared samples (n = 15) in the range of 0-3.0% and analyzed on the EPICS-XL flow cytometer.
7
In study site #1, the same prepared samples (n =50) were analyzed on both the Becton Dickinson Facscan and Coulter EPICS-XL flow cytometers, as follows
Summary of Correlations on FACscan Flow Cytometer Prepared Samples
| Comparisons | Mean %
Positive | Confidence
Interval (95%) | r2
value | n |
|----------------------|--------------------|------------------------------|-------------|----|
| HbF FITC
KB | 4.54
4.41 | 3.63
3.52 | 97.95 | 50 |
| HbF R-PE
KB | 4.50
4.41 | 3.59
3.52 | 98.16 | 50 |
| HbF TC
KB | 4.49
4.41 | 3.58
3.52 | 97.98 | 50 |
| HbF FITC
HbF R-PE | 4.54
4.50 | 3.63
3.59 | 99.59 | 50 |
| HbF FITC
HbF TC | 4.54
4.49 | 3.63
3.58 | 99.63 | 50 |
| HbF R-PE
HbF TC | 4.50
4.49 | 3.59
3.58 | 99.70 | 50 |
Summary of Correlations on EPICS-XL Flow Cytometer Prepared Samples
| Comparisons | Mean %
Positive | Confidence
Interval (95%) | r2 Value | n |
|----------------------|--------------------|------------------------------|----------|----|
| HbF FITC
KB | 4.22
4.40 | 3.38
3.52 | 98.48 | 50 |
| HbF R-PE
KB | 4.24
4.40 | 3.40
3.52 | 98.41 | 50 |
| HbF TC
KB | 4.20
4.40 | 3.36
3.52 | 97.96 | 50 |
| HbF FITC
HbF R-PE | 4.22
4.24 | 3.38
3.40 | 99.61 | 50 |
| HbF FITC
HbF TC | 4.22
4.20 | 3.38
3.36 | 99.54 | 50 |
| HbF R-PE
HbF TC | 4.24
4.20 | 3.40
3.36 | 99.67 | 50 |
In the following linear regression analyses, the correlations of the Caltag Fetal Hemoglobin test and KB test are presented separately for each study site with prepared and patient samples identified within each site. However, prepared samples analyzed in study site #1 are as indicated in the above tables.
8
| Site 1
| HAF FITC | Patient Samples | FACscan Flow Cytometer | |||
|---|---|---|---|---|---|
| procedure | mean % | ||||
| positive | Confidence | ||||
| Interval (95%) | |||||
| ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | r 2 | ||||
| value | ന | ||||
| HbF FITC | |||||
| KB Method | 0.51 | ||||
| 0.51 | 0.04 | ||||
| 0.03 | 96.80 | 30 | |||
| HbF R-PE | Patient Samples | ||||
| procedure | positive | mean % Confidence | |||
| Interval (95%) | r2 | ||||
| value | റ | ||||
| HbF R-PE | |||||
| KB Method | 0.50 | ||||
| 0.51 | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | ||||
| 0.05 | |||||
| 0.03 | 96.84 | 30 | |||
| HPF TC | |||||
| procedure | Patient Samples | ||||
| mean % | |||||
| positive | Confidence | ||||
| Interval (95%) | r2 | ||||
| value | n | ||||
| HbF TC | |||||
| KB Method | 0.47 | ||||
| 0.51 | ============================================================================================================================================================================== | ||||
| 0.05 | |||||
| 0.03 | 97.50 30 | ||||
| Site 2 | |||||
| HbF FITC | Patient Samples | EPICS-XL Flow Cytometer | and and the comments of the comments of the comments of | ||
| procedure | mean % | ||||
| positive | Confidence | ||||
| Interval (95%) | r2 | ||||
| value | n | ||||
| HbF FITC | |||||
| KB Method | 0.22 | ||||
| 0.20 | +============================================================================================================================================================================ | ||||
| 0.07 | |||||
| 0.05 | 98.34 | 38 | |||
| HPF FITC | Prepared Samples | EPICS-XL Flow Cytometer | |||
| procedure | mean % | ||||
| positive | Confidence | ||||
| Interval (95%) | 12 | ||||
| value | n | ||||
| HBF FITC | |||||
| KB Method | 1.52 | ||||
| 1.61 | ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------- | ||||
| 1.04 | |||||
| 1 . 1 1 | 85.50 15 | ||||
| . Site 3 | |||||
| HPF R-PE | Patient Samples | EPICS-XL Flow Cytometer | |||
| procedure | mean % | ||||
| positive | Confidence | ||||
| Interval (95%) | r2 | ||||
| value | n | ||||
| HbF R-PE | 0.08 | ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | |||
| 9.43 | 64.00 | 13 | |||
| KB Method | 0.11 | 0.01 | ﺎﻧ | ||
| HbF R-PE | Prepared Samples | EPICS-XL Flow Cytometer | |||
| procedure | mean % | ||||
| positive | Confidence | ||||
| Interval (95%) | r2 | ||||
| value | រា | ||||
| HbF R-PE | |||||
| KB Method | 1.38 | ||||
| 1.40 | ============================================================================================================================================================================== | ||||
| 0.93 | |||||
| 0.81 | 84.01 | 15 |
.
9
BIBLIOGRAPHY
- Freedman J., Lazarus A.H., Applications of flow cytometry in transfusion medicine, Transfusion 1. Medicine Reviews 9:87-109, 1995.
- Davis B. H., Flow Cytometric Analysis of Red Blood Cells in Clinical Flow Cytometry: Principles and 2. Application, pp 373-387, Williams & Wilkins, 1993.
- Sebring E.S., Polesky H.F., Fetomaternal hemorrhage: insk factors, ume of occurrence and 3. clinical effects, Transfusion 30:344-357, 1990.
- Kleihauer E., Braun H., Betke K., Demonstration of fetal hemoglobin in erythrocyces of a blood smear. 4. Klin. Wochenschr. 35:637-638, 1957.
- Emery C.L., Morway L.F., Chung-Park M. et. al., The Kleihauer-Betke Test, Clinical utility, న్. indication, and correlation in patients with placental abruption and cocaine use, Arch. Pathol. Lab. Med. 119:1032-1037, 1995.
- Towery R., English T.P., Wisner D., Evaluation of pregnant women after blunt injury, J. Trauma 6. 35:731-736, 1993.
- Corsetti J.P., Cox C., Leary J.F. et. al., Comparison of quantitative acid-elution technique and flow cytometry for 7. detecting fetal maternal hemorrhage, Ann. Clin. Lab Sci., 17:197-206. 1987.
- Nance S.J., Nelson J.M., Arnot P.A., Quantiation of fetal-maternal hemorrhage by flow cytomerry, Am. J. Clin. 8. Path. 91:288-292, 1989.
-
- Nance S.J., Garraty G., Application of flow cytometry to immunohematology, J. Immunol. Meth. 101:127-131. 1987.
-
- Medaris A.L., Hensleigh P.A., Parks D.R. et. al., Detection of fetal erythrocytes in maternal blood post partum with the fluorescence-activated cell sorter, Am. J. Obstet. Gynecol. 148(3):290-295, 1984.
- Thorne S.J., Thein S.L., Sampierro M. et. al., Immunochemical estimation of hemoglobin types in red blood cells 11. by FACS analysis, Brit. J. Hemat. 87:125-132, 1994.
- Davis B.H., Olsen S., Bigelow N.C. et. al., Detection of fetal red cells in fetomaternal hemorrhage using an ani-12. hemoglobin F monoclonal antibody by flow cytometry, Transfusion 38:749-756,1998.
- Stamatoyannopoulos G., Nienhuis A.W., Hemoglobin switching, in The Molecular Basis of Blood Diseases, pp. 67-13. 105. Saunders & Co., Philadelphia, 1987.
-
- Nicholson J.K.A., Green T.A. and Collaborative Laboratories, Selection of anticoagulants for Ivmphocyte immunophenotyping. J. Immunol, Methods 165:31-35. 1993.
-
- Brown M.C., Hoffman R.A., Kirchanski S., Controls for flow cytometers in hematology and cellular immunology, Ann. N.Y. Acad. Sci. 468:93-103, 1986.
-
- Durrand R.E., Calibration of flow cytometer detector systems, Cytometry 2:192-193, 1981.
10
Image /page/10/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized caduceus symbol, which is a staff with two snakes coiled around it, representing medicine and healing. The symbol is surrounded by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" in a circular arrangement. The text is in all capital letters and is evenly spaced around the symbol.
Food and Drug Administration 2098 Gaither Road Rockville MD 20850
SEP 1 7 1999
Caltag Laboratories c/o David C. Bishop, Ph.D. 605 Dilworth Road Downingtown, Pennsylvania 19335
Re: K990641 Trade Name: Caltag Fetal Hemoglobin Test Regulatory Class: II Product Code: GHQ Dated: July 13, 1999 Received: July 14, 1999
Dear Dr. Bishop:
We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.
If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.
11
Page 2
Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770) 488-7655.
This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled. "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597, or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".
Sincerely yours.
Steven Butman
Steven I. Gutman, M.D. M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health
Enclosure
12
| Page | of |
|---|---|
| Comments of the consisted on any and the consisted to the contribution of the contribution of the contribution of the contribution of the contribution of the contribution of | |
| ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ |
510(k) Number (if known): _ K 990641
Ocvice Name:_CALIAG_FETAL HEMOGGOON_TEST
Indications For Use:
The Caltag Fetal Hemoglobin Test, containing either FITC, R-PE or TRI-COLOR® The Cattag Petal Hemoglobin Toot, Contractor Coating Cobin F), is intended for confugation followed by enumeration of fetal red blood cells. Fetal red cells are identified by the presence of feal hemoglobin by a flow cytometric method. Fetal cells, when found in the maternal circulation, may be an indication of fetal or maternal Cells, when round in the nateromplications. The hemorrhage of Rh+ fetal blood into trauna of various obsited complex of sensitizing Rh antibodies in the RIF materilar biood may readle in the edministration to the mother of Rh mother. Sellsluzation may or proveneed of a The Caltag Fetal Hemoglobin Test may millianc globanii (ruirg) soon all incompatible fetal-maternal hemorrhage and be used as an all in detecting "immunoprophylaxis with Rh immune globulin.
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)
Concurrence of CORH, Office of Device Evaluation (ODE)
(Division Sign-Off)
Division of Clinical Laboratory Devices K990641
510(k) Number
Prescription Use
(Per 21 CFR 801.109)
OR
Over-The-Counter Use_
(Optional Format 1-2-96)