The CAPI 3 HEMOGLOBIN(E) kit is designed for the separation of the normal hemoglobins (A. A2 and F) in human venous blood samples, and for the detection of the major hemoglobin variants (S, C, E and D), by capillary electrophoresis in alkaline buffer (pH 9.4) with the SEBIA CAPILLARYS 3 TERA instrument.

The CAPILLARYS 3 TERA instrument is an automated analyzer which performs a complete for the quantitative analysis of the normal hemoglobin fractions A. A2 and F and for themoglobin variants S. C. E and D. The assay is performed on the hemolysate of whole blood samples collected in tubes containing K2EDTA or K3EDTA as anticoagulant. The CAPI 3 HEMOGLOBIN(E) is intended to be used in conjunction with other laboratory and clinical findings.

For In Vitro Diagnostic Use

Device Description

The CAPILLARYS 3 instrument uses the principle of capillary electrophoresis in free solution which is the most common form of capillary electrophoresis. With this technique, charged molecules are separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation also occurs according to the electrolyte pH and electroosmotic flow.

The CAPILLARYS 3 instrument has silica capillaries functioning in parallel allowing 12 simultaneous analyses for hemoglobin quantification in a whole blood sample. A sample dilution with hemolysing solution is prepared and injected by aspiration at the anodic end of the capillary. A high voltage protein separation is then performed and direct detection of the hemoglobins is made at the cathodic end of the capillary at 415 nm, which is the absorbance wave length specific to hemoglobins. Before each run, the capillaries are washed with a wash solution and prepared for the next analysis with buffer.

By using alkaline pH buffer, normal and abnormal (or variant) hemoglobins are detected in the following order, from cathode to anode: δΑ'2 (A2 variant). C. A2, E. S. D. F. and A.

The carbonic anhydrase is not visualized on the hemoglobin electrophoretic patterns by capillary electrophoresis, this permits to identify hemoglobin A2 variants in this migration zone.

NOTE : the name "CAPILLARYS 3" is used for the SEBIA CAPILLARYS 3 TERA automated instrument.

The hemoglobins are reported in % units along with an electrophoresis scan.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study that proves the device meets them, based on the provided FDA 510(k) summary:

Device: CAPI 3 HEMOGLOBIN(E) kit used with SEBIA CAPILLARYS 3 TERA instrument.

Intended Use: For the separation of normal hemoglobins (A, A2, F) in human venous blood and for the detection of major hemoglobin variants (S, C, E, D) by capillary electrophoresis. It provides quantitative analysis of fractions A, A2, F and detection of variants S, C, E, D.

1. Table of Acceptance Criteria and Reported Device Performance

The document doesn't explicitly state "acceptance criteria" in a separate section with specific numerical thresholds for all performance metrics. Instead, it demonstrates performance through various studies and implies that the observed performance (e.g., correlation coefficients close to 1, low CVs for precision) is acceptable for substantial equivalence to a predicate device.

However, based on the provided performance data, we can infer the implied acceptance criteria from the reported excellent results, particularly for method comparison:

Performance Metric	Implicit Acceptance Criteria (Inferred from reported data)	Reported Device Performance
Precision/Reproducibility	Low coefficients of variation (CV%) for all hemoglobin fractions (Hb A, Hb A2, Hb F, Hb S, Hb C, Hb D, Hb E) across within-run, between-run, between-day, between-instrument, and total reproducibility studies.	7-days Reproducibility (3 instruments, 1 lot):- Hb A Total CV: 0.0% - 1.3%- Hb A2 Total CV: 1.3% - 6.5%- Hb F Total CV: 2.5%- Hb S Total CV: 0.6%- Hb C Total CV: 1.4%- Hb D Total CV: 1.4%- Hb E Total CV: 1.6%20-days Reproducibility (1 instrument, 1 lot):- Hb A Total CV: 0.0% - 0.8%- Hb A2 Total CV: 1.4% - 6.0%- Hb F Total CV: 0.7%- Hb S Total CV: 1.0%- Hb C Total CV: 1.7%- Hb D Total CV: 0.6%- Hb E Total CV: 1.1%
Linearity	Demonstrated linearity across the clinically relevant range for all hemoglobin fractions.	Determined to be linear within the entire range studied for: - Hb A (1.0 - 97.3%)- HbS (1.1 - 89.7%)- Hb A2 (0.2 - 9.1%)- Hb F (0.5 - 83.1%)- Hb C (0.3 - 82.0%)- Hb D (1.1 - 43.5%)- Hb E (0.3 - 86.9%)
Limit of Blank (LOB)	Very low LOB values.	Hb A, Hb A2, Hb F, Hb S, Hb C, Hb D, Hb E: All 0.1% or 0.2%
Limit of Detection (LOD)	Low LOD values indicating sensitivity to detect low concentrations.	Hb A: 1.0%, Hb A2: 0.2%, Hb F: 0.4%, Hb S: 0.9%, Hb C: 0.3%, Hb D: 0.7%, Hb E: 0.3%
Limit of Quantitation (LOQ)	Low LOQ values indicating ability to accurately quantify at low concentrations.	Hb A: 1.0%, Hb A2: 0.2%, Hb F: 0.5%, Hb S: 1.1%, Hb C: 0.3%, Hb D: 1.1%, Hb E: 0.3%
Analytical Specificity (Interference)	Insignificant interference from common substances like bilirubin and triglycerides at elevated levels.	Interference studies showed that Bilirubin (20.6 mg/dL) and Triglycerides (2.2 g/dL) did not significantly interfere with the analytical performance. (The document states studies were "conducted" and lists maximum concentrations without giving specific results of non-interference, but the conclusion implies acceptability).
Method Comparison (Correlation)	Very high correlation coefficients (close to 1.000) for all hemoglobin fractions and variants when compared to a reference method, along with slopes near 1 and y-intercepts near 0 in regression analysis, demonstrating strong agreement.	Site 1: - Hb A: 1.000 (Slope ~1.01, Y-intercept ~-1.0 to -0.7) - Hb A2: 0.998 (Slope ~1.00, Y-intercept ~0.0 to -0.05) - Hb F: 1.000 (Slope ~1.00-1.01, Y-intercept ~-0.008 to 0.05) - Hb S, C, D, E: All 1.000 (Slopes ~1.01-1.02, Y-intercepts close to 0) Site 2: - Hb A: 1.000 (Slope ~1.01-1.02, Y-intercept ~-1.9 to -1.4) - Hb A2: 0.987 (Slope ~1.00-1.01, Y-intercept ~0.0 to -0.005) - Hb F: 0.999 (Slope ~0.96-1.00, Y-intercept ~-0.12 to 0.0) - Hb S, C, E: All 0.997 - 1.000 (Slopes ~1.00-1.06, Y-intercepts close to 0 or small variations)
False Positives (Variant Detection)	No false positives in the detection of abnormal hemoglobin bands or abnormal levels of normal bands.	"There was no case observed of false positive, i.e., detection of an abnormal band or abnormal level of a normal band where no such abnormality existed." This statement is made for both Site 1 and Site 2 studies.

2. Sample Size Used for the Test Set and Data Provenance

Sample Size for Test Set (Method Comparison): A total of 304 samples were used across two sites.
- Site 1: 153 samples (64 with hemoglobin variants)
- Site 2: 151 samples (60 with hemoglobin variants)
Data Provenance: The samples were "provided by hospitals and laboratories international and United States." The study design is implied to be retrospective, as samples were collected and then analyzed by both the candidate and reference methods.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications

The document does not explicitly state the number of experts used to establish ground truth or their specific qualifications (e.g., "radiologist with 10 years of experience").
The "reference procedure" used for comparison is described as "a commercially available capillary electrophoresis technique for hemoglobin analysis." This implies that the ground truth for quantitative values and variant identification was established by a previously validated and accepted clinical laboratory method. For abnormal hemoglobin detection, the agreement with "clinical diagnosis" is also mentioned, suggesting that expert clinical assessment contributed to the overall understanding of the ground truth.

4. Adjudication Method (for the test set)

The document does not explicitly describe an adjudication method (e.g., 2+1, 3+1).
The comparison studies directly compare the quantitative results and variant detection of the candidate device against a "reference procedure." Any discrepancies would typically be investigated, but the method for their resolution is not detailed. The statement of "no observed false positives" implies a direct agreement determination.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC comparative effectiveness study was not done.
This device is an in-vitro diagnostic (IVD) for laboratory use that provides quantitative measurements and detects specific analytes. It does not involve human readers interpreting images or data for diagnosis in a way that would typically warrant an MRMC study to assess AI-assisted human performance improvement. The performance is assessed by comparison to a reference laboratory method.

6. Standalone Performance Study (algorithm only without human-in-the-loop performance)

Yes, the provided performance data primarily represents the standalone performance of the device.
The CAPI 3 HEMOGLOBIN(E) kit processes samples and provides quantitative results and variant detection via the automated CAPILLARYS 3 TERA instrument. The precision, linearity, LOB/LOD/LOQ, and analytical specificity studies, as well as the method comparison studies, evaluate the device's inherent analytical performance without direct human intervention in the result generation or interpretation (beyond standard laboratory procedures for running the instrument and reviewing results, which is inherent to any IVD). The measurements and variant identifications are generated by the instrument's software.

7. Type of Ground Truth Used

The ground truth for the test set was established primarily through:

Reference Procedure/Comparative System: A "commercially available capillary electrophoresis technique for hemoglobin analysis" (predicate device or similar accepted method). This acts as the gold standard for quantitative values and identification of hemoglobin fractions and variants.
Clinical Diagnosis: For the detection of abnormal hemoglobins, agreement with "clinical diagnosis" is also mentioned, suggesting that patient medical records and expert clinical assessment contributed to confirming the presence or absence of variants.

8. Sample Size for the Training Set

The document does not explicitly state the sample size for the training set.
As this is a 510(k) submission for an IVD kit and instrument, it's likely that extensive internal development and validation data would have been generated during the device's creation (which can be likened to a training/development phase), but this specific information is not typically required in the 510(k) summary provided. The focus of the 510(k) is often on the clinical validation/test set performance.

9. How the Ground Truth for the Training Set Was Established

The document does not explicitly describe how the ground truth for a training set was established.
Given that this is an analytical device, the "training" (development) process would involve optimizing reagents, instrument parameters, and algorithms to accurately measure hemoglobin fractions and identify variants. This would typically involve using well-characterized control materials, spiked samples, and patient samples with known hemoglobin profiles (established by reference methods, genetic testing, or clinical diagnosis) to calibrate and refine the system, but the specifics are not detailed in this summary.

Summary

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December 14, 2018

Sebia, Inc. Karen Anderson Director of Technical & Regulatory 1705 Corporate Drive, Suite 400 Norcross, Georgia 30093

Re: K180762

Trade/Device Name: CAPI 3 HEMOGLOBIN(E) Regulation Number: 21 CFR 864.7415 Regulation Name: Abnormal hemoglobin assay Regulatory Class: Class II Product Code: GKA Dated: March 23, 2018 Received: March 23, 2018

Dear Karen Anderson:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and 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) that do not require approval of a premarket approval application (PMA). You mav, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be avare that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you; however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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requirements, including, but not limited to: registration and listing (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Leonthena R. Carrington -S

Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K180762

Device Name CAPI 3 HEMOGLOBIN(E)

Indications for Use (Describe)

For In Vitro Diagnostic Use

Type of Use (Select one or both, as applicable)

Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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510K SUMMARY (Summary of Safety and Effectiveness)

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

Submitter Name	Sebia, Inc.
Address	1705 Corporate Drive Suite 400Norcross, Georgia 30093, USA
Contact	Karen Anderson, Dir of Technical and RegulatoryPhone: 1-800-835-6497, 3704Fax: 770-446-8511Email: karen.anderson@sebia-usa.comMatthew C. Wagner, Ph.D. ScientificAffairs SpecialistPhone: 1-800-835-6497Email: matthew.wagner@sebia-usa.com
Date Prepared	December 13, 2018
Manufacturing	SebiaParc Technologique Léonard de VinciRue Léonard de Vinci,CP 8010 LISSES, 91008 EVRY CedexFRANCEPhone: (33) 1 69 89 80 80Fax: (33) 1 69 89 78 78
Product Name	CAPI 3 HEMOGLOBIN(E)
Common Name	Hemoglobin by capillary electrophoresis
Product Regulation No.	21 CFR 864.7415
Product Codes , Deviceclassification and PanelClassification	GKA, Class II , Hematology (81)
Establishment Registration No.	8023024

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1. DEVICE DESCRIPTION

By using alkaline pH buffer, normal and abnormal (or variant) hemoglobins are detected in the following order, from cathode to anode: δΑ'2 (A2 variant). C. A2, E. S. D. F. and A.

The carbonic anhydrase is not visualized on the hemoglobin electrophoretic patterns by capillary electrophoresis, this permits to identify hemoglobin A2 variants in this migration zone.

NOTE : the name "CAPILLARYS 3" is used for the SEBIA CAPILLARYS 3 TERA automated instrument.

The hemoglobins are reported in % units along with an electrophoresis scan.

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Reagents:

CAPI 3 HEMOGLOBIN(E) KIT

ITEMS	PN 2507
Buffer (ready to use)	2 vials, 700 mL each
Hemolysing solution (ready to use)	1 vial, 700 mL
Filters	4 filters

Additional reagents and accessories not included in the CAPI 3 HEMOGLOBIN(E) KIT

ITEMS	PN	COMPONENTS
CAPICLEAN CAPILLARYS 3	2060	1 vial, 25 mL
CAPILLARYS 3 WASH SOLUTION	2062	1 vial, 75mL
CAPI 3 REAGENT CUPS	2582	24 X 14 packs of reagent cups
TEST TUBES	9214	200 of 100 mm-tubes
CAPI 3 BINS	2581	5 units
TUBES AND CAPS FOR	9202	20 units
CONTROLS	9205	500 units
CAPILLARYS 3 & MC SWITCHRACK FOR HEMOGLOBIN(E)	1373	1 unit
CAPILLARYS 3 & MC LOWVOLUME RACKS	1364	5 units
SEBIA CAPILLARYS 3	1246	1 unit

2. INDICATIONS FOR USE

CAPI 3 HEMOGLOBIN(E) kit:

The CAPI 3 HEMOGLOBIN(E) kit is designed for the separation of the normal hemoglobins (A, A2 and F) in human venous blood samples, and for the detection of the major hemoglobin variants (S. C. E and D), by capillary electrophoresis in alkaline buffer (pH 9.4) with the SEBIA CAPILLARYS 3 TERA instrument.

The CAPILLARYS 3 TERA instrument is an automated analyzer which performs a complete hemoglobin profile for the quantitative analysis of the normal hemoglobin fractions A, A2 and F and for the detection of major hemoglobin variants S, C, E and D. The assay is performed on the hemolysate of whole blood samples collected in tubes

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containing K₂EDTA or K₃EDTA as anticoagulant. The CAPI 3 HEMOGLOBIN(E) is intended to be used in conjunction with other laboratory and clinical findings.

For In Vitro Diagnostic Use.

3. TECHNOLOGICAL CHARACTERISTICS

The CAPILLARYS 3 instrument in combination with the CAPI3 HEMOGLOBIN(E) kit uses the principle of capillary electrophoresis in free solution which is the most common form of capillary electrophoresis. With this technique, charged molecules are separated by their electrophoretic mobility in an alkaline buffer with a specific pH. Separation also occurs according to the electrolyte pH and electroosmotic flow.

Direct detection provides accurate relative quantification of individual hemoglobin fraction, and the resulting electrophoregrams are also evaluated visually for pattern abnormalities. In addition, the high resolution of this procedure should allow the identification of hemoglobin variants, in particular, to differentiate hemoglobins S from D, and E from C. The hemoglobin A2 quantification can also be performed when hemoglobin E is present. A2 hemoglobin quantification may be used with other clinical and laboratory findings for ß thalassemia detection.. By using alkaline pH buffer, normal and abnormal (or variant) hemoglobins are detected in the following order, from cathode to anode: δΑ΄2 (A2 variant), C, A2, E, S, D, F and A. The carbonic anhydrase is not visualized on the hemoglobin electrophoretic patterns by capillary electrophoresis, this permits to identify hemoglobin A2 variants in this migration zone.

4. SUBSTANTIAL EQUIVALENCE INFORMATION:

Predicate Device Name	Predicate Device 510(k) number	Product Code	Regulation No.
CAPILLARYS HEMOGLOBIN(E) using theCAPILLARYS 2 FLEX-PIERCINGinstrument,	K112550	GKA	864.7415

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Similarities between the candidate device (CAPI 3 HEMOGLOBIN(E)) and the predicate device (CAPILLARYS HEMOGLOBIN(E)), K112550 (Table A).

Similarities
Table A	Sebia CAPI 3 HEMOGLOBIN(E)Candidate Device	Sebia CAPILLARYSHEMOGLOBIN(E)Predicate Device (K112550)
Intended use	The CAPI 3 HEMOGLOBIN(E) kit isdesigned for the separation of the normalhemoglobins (A, A2 and F) in human venousblood samples, and for the detection of themajor hemoglobin variants (S, C, E and D), bycapillary electrophoresis in alkaline buffer (pH9.4) with the SEBIA CAPILLARYS 3 TERAinstrument.The CAPILLARYS 3 TERA instrument is anautomated analyzer which performs acomplete hemoglobin profile for thequantitative analysis of the normalhemoglobin fractions A, A2 and F and for thedetection of major hemoglobin variants S, C,E and D. The assay is performed on thehemolysate of whole blood samples collectedin tubes containing K2EDTA or K3EDTA asanticoagulant. The CAPI 3 HEMOGLOBIN(E)is intended to be used in conjunction withother laboratory and clinical findings.For In Vitro Diagnostic Use.	The CAPILLARYS HEMOGLOBIN(E)kit is designed for the separation ofthe normal hemoglobins (A, A2 and F)in human blood samples, and for thedetection of the major hemoglobinvariants (S, C, E and D), by capillaryelectrophoresis in alkaline buffer (pH9.4) with the SEBIA CAPILLARYS 2FLEX-PIERCING instrument.The CAPILLARYS 2 FLEXPIERCINGinstrument is an automated analyzerwhich performs a completehemoglobin profile for the quantitativeanalysis of the normal hemoglobinfractions A, A2 and F and for thedetection of major hemoglobinvariants S, C, E and D. The assay isperformed on the hemolysate of wholeblood samples collected in tubescontaining K2EDTA or K3EDTA asanticoagulant.For In Vitro Diagnostic Use.
Specimen Type	Venous Human Whole Blood	Same
Technology	CAPILLARYS ELECTROPHORESIS	Same
DETECTIONAbsorbanceWavelength	415 nm	Same
Software	PHORESIS	Same
BarcodeIdentification ofSame	On-board	Same
Controls formigration	Sebia Normal A2 Control ( sold separately)	Same
Buffer andComposition	CAPILLARYS HEMOGLOBIN(E)	Same
Use of BufferSolution	On-Board	Same
Wash Solutionand Composition	Same	Same
Use of the WashSolution	On-Board	Same
HemolysingSolution andComposition	On-Board	Same
Use ofHemolysingSolution	On-Board	Same
Reference Range	Hb A 96.7-97.8%HbF ≤ 0.5 %Hb A2 2.2-3.2 %	Same
Hb variants library(on-board)	Yes, displayed by the software and indicatedin the package insert)	Same

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Table B. Differences between the candidate device (CAPI 3 HEMOGLOBIN(E)) and the predicate device (CAPILLARYS HEMOGLOBIN(E), K112550 in (Table B).

Differences
Table B	Sebia CAPI 3 HEMOGLOBIN(E)Candidate Device	Sebia CAPILLARYS HEMOGLOBIN(E)Predicate Device (K112550)
Number of Separationunits ( Capillaries)	12	8
Bottle for reagents	RFID tag	None
Reagent Cups	Supplied separate packaging	Supplied in the kit
Wash Solution	Supplied separate packaging	Supplied in the kit

5. Performance Data:

a. Precision / Reproducibility:-

The precision of the CAPI 3 HEMOGLOBIN(E) procedure was evaluated in studies based on the Clinical and Laboratory Standards Institute (CLSI - USA) EP5-A3 guideline "Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline – Third Edition". The means, standard deviations (SD) and coefficients of variation (CV %) were calculated for percentage (%) of hemoglobin fractions for each sample.

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7-days reproducibility study with three instruments and one lot of kit

Seven (7) different native blood samples were run using the CAPI 3 HEMOGLOBIN(E) procedure performed with three CAPILLARYS 3 TERA instruments.

The 7-days reproducibility performed with three CAPILLARYS 3 instrument and one lot of CAPI 3 HEMOGLOBIN(E) kit is summarized in the following within-run, between-run, between-day, between-instrument and total reproducibility precision estimates (SD and % CV ranges) for the percentages (%) of each hemoglobin fraction from all samples.

Ranges of % tested			Within-run				Between-run				Between-day
Fraction	Min Value	Max Value	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max
Hb A	56,5	97,5	0,03	0,39	0,0%	0,6%	0,00	0,22	0,0%	0,4%	0,00	0,73	0,0%	1,3%
Hb A2	1,9	4,8	0,03	0,14	1,1%	5,2%	0,00	0,05	0,0%	1,6%	0,00	0,11	0,0%	3,8%
Hb F		34,9	0,15		0,4%		0,08		0,2%		0,84		2,4%
Hb S		40,1	0,14		0,4%		0,06		0,1%		0,10		0,2%
Hb C		34,3	0,36		1,1%		0,11		0,3%		0,28		0,8%
Hb D		38,6	0,15		0,4%		0,11		0,3%		0,50		1,3%
Hb E		23,5	0,18		0,8%		0,00		0,0%		0,32		1,4%

CAPI 3 HEMOGLOBIN(E) & CAPILLARYS 3 TERA : Reproducibility study on native whole blood samples (2018/10)

Ranges of % tested			Between-instrument				Total reproducibility(*)
Fraction	Min Value	Max Value	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max
Hb A	56,5	97,5	0,00	0,14	0,0%	0,2%	0,03	0,84	0,0%	1,3%
Hb A2	1,9	4,8	0,00	0,03	0,0%	0,5%	0,03	0,17	1,3%	6,5%
Hb F		34,9	0,00		0,0%			0,86		2,5%
Hb S		40,1	0,16		0,4%			0,24		0,6%
Hb C		34,3	0,00		0,0%			0,47		1,4%
Hb D		38,6	0,00		0,0%			0,54		1,4%
Hb E		23,5	0,00		0,0%			0,37		1,6%

(*) Total reproducibility includes : within-run, between-run, between-day and between-instrument.

20-days reproducibility study with one instrument and one lot of kit

Five (5) different samples were run using the CAPI 3 HEMOGLOBIN(E) procedure performed with one CAPILLARYS 3 instrument and one lot on CAPI 3 HEMOGLOBIN(E) kit. Each sample was analyzed in duplicate on twelve capillaries per run, two runs per day over 20 days yielding a total of 960 results per sample.

The 20-days reproducibility performed with one CAPILLARYS 3 instrument and one lot of CAPI 3 HEMOGLOBIN(E) kit is summarized in the following tables including within-capillary, betweencapillary, between-run, between-day and total reproducibility precision estimates (SD and % CV ranges) for the percentages (%) of each hemoglobin fraction from all samples

Fraction	Ranges of % tested		Within-capillary				Between-capillary				Between-run
	Min Value	Max Value	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max
Hb A	44,2	97,3	0,04	0,31	0,0%	0,5%	0,01	0,16	0,0%	0,4%	0,00	0,14	0,0%	0,3%
Hb A2	2,6	6,5	0,04	0,09	0,8%	3,1%	0,02	0,12	0,9%	4,4%	0,00	0,06	0,0%	2,4%
Hb F		26,8		0,12		0,4%		0,02		0,1%		0,14		0,5%
Hb S		17,5		0,06		0,4%		0,08		0,5%		0,02		0,1%
Hb C		8,9		0,08		0,9%		0,07		0,8%		0,04		0,4%
Hb D		40,6		0,19		0,5%		0,06		0,2%		0,05		0,1%
Hb E		22,6		0,23		1,0%		0,02		0,1%		0,09		0,4%

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Fraction	Ranges of % tested		Between-day				Total reproducibility(*)
	Min Value	Max Value	SD min	SD max	CV min	CV max	SD min	SD max	CV min	CV max
Hb A	44,2	97,3	0,02	0,22	0,0%	0,5%	0,05	0,40	0,0%	0,8%
Hb A2	2,6	6,5	0,02	0,05	0,7%	1,3%	0,05	0,16	1,4%	6,0%
Hb F		26,8	0,07		0,2%		0,19		0,7%
Hb S		17,5	0,13		0,7%		0,17		1,0%
Hb C		8,9	0,10		1,1%		0,15		1,7%
Hb D		40,6	0,12		0,3%		0,24		0,6%
Hb E		22,6	0,06		0,3%		0,26		1,1%

(*) Total reproducibility includes : within-capillary, between-capillary, between-run and between

b. Linearity

A linearity study was performed per CLSI EP06-A: Evaluation of Quantitative Measuring Procedures; A Statistical Approach. The results for percentage (%) of hemoglobin fractions were analyzed using statistical tools recommended by CLSI.

Mixtures of two different blood samples were mixed at different proportions and tested in triplicate and analyzed using the CAPI 3 HEMOGLOBIN(E) procedure on the CAPILLARYS 3 instrument. The tests were determined to be linear within the entire range studied for each of the following hemoglobins fractions:

Hb A ( 1.0 -97.3%), HbS (1.1-89.7%), Hb A2 (0.2-9.1%), Hb F (0.5-83.1%), Hb C (0.3-82.0%), Hb D (1.1-43.5%), Hb E (0.3-86.9%).

c. Limit of Blank (LOB), Limit of Detection (LOD), Limit of Quantitation (LOQ)

Per CLSI guidelines , EP17-A, Protocols for Determination of Limits of Detection and Limits of Quantitation , studies were conducted using the CAPI 3 HEMOGLOBIN(E) procedure using the CAPILLARYS 3 for each hemoglobin fraction using five (5) different blood samples. Results are as follows:

Fraction	LOB %	LOD %	LOQ %
Hb A	0,1	1,0	1,0
Hb A2	0,1	0,2	0,2
Hb F	0,2	0,4	0,5
Hb S	0,1	0,9	1,1
Hb C	0,1	0,3	0,3
Hb D	0,1	0,7	1,1
Hb E	0,1	0,3	0,3

d. Analytical Specificity

Interference studies were conducted following CLSI, EP7-A2, Interference Testing in Clinical Chemistry. Three (3) blood samples (one blood sample with normal Hb A2 level, one blood sample with increased Hb A2 level and one blood sample with Hb S). Each sample was analyzed 3 times for reproducibility using CAPI 3 HEMOGLOBIN(E) procedure and CAPILLARYS 3 TERA instrument :

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Interferents	Maximum Concentration
Bilirubin	20.6 mg/dL , or 352 µmol/L
Triglycerides	2.2 g/dL , or 25.1 mmol/L

e. Comparison Studies

Method comparison studies were preformed using CAPI 3 HEMOGLOBIN(E) assay using the CAPILLARYS 3 TERA instrument. The studies were conducted following CLSI, EP09-A2-IR, Method Comparison and Bias Estimation Using Patient Samples-200 edition.

The samples were provided by hospitals and laboratories international and United States. A total of 304 samples (180 without hemoglobin variant /124 with hemoglobin variant) were analyzed.

The measured values of hemoglobin fractions from both procedures were analyzed by three regression statistical procedures. The results of regression analysis are tabulated below:

Site 1:

The levels of hemoglobin fractions were measured in 153 blood samples, including 64 samples with hemoqlobin variants, both by electrophoretic separations obtained with the CAPI 3 HEMOGLOBIN(E) procedure performed with the CAPILLARYS 3 instrument and a commercially available capillary electrophoresis technique for hemoglobin analysis (reference).

Fraction	Number of samples	Correlation coefficient	Ordinary linear regression		Weighted Deming regression		Passing-Bablok regression		Range of Hb % valuesCAPI 3 HEMOGLOBIN(E)
			y-intercept	Slope	y-intercept	Slope	y-intercept	Slope
Hb A	150	1.000	-0.993	1.010	-0.703	1.007	-0.994	1.010	16.9 - 98.7
Hb A2	148	0.998	0.005	0.986	-0.032	1.000	-0.050	1.000	0.5 - 9.2
Hb F	22	1.000	-0.008	1.009	0.049	0.999	0.027	1.009	0.8 - 83.1
Fraction	Number of samples	Correlation coefficient	Ordinary linear regression		Weighted Deming regression		Passing-Bablok regression		Range of Hb % valuesCAPI 3 HEMOGLOBIN(E)
			y-intercept	Slope	y-intercept	Slope	y-intercept	Slope
Hb S	13	1,000	-0,025	1,010	-0,127	1,013	-0,122	1,013	1,8 - 89,7
Hb C	13	1,000	0,099	1,008	0,009	1,009	-0,163	1,018	2,0 - 89,5
Hb D	9	1,000	-0,068	1,015	0,063	1,008	-0,032	1,015	3,3 - 43,7
Hb E	13	1,000	0,183	1,001	-0,054	1,015	-0,080	1,020	5,0 - 86,9

Normal hemoglobins

This study demonstrated a perfect correlation between the 2 analysis procedures for the Hb A, Hb A2, Hb F, Hb S, Hb C, Hb D and Hb E quantitative determination.

All abnormal hemoglobins or abnormal levels of normal hemoglobins detected with the CAPI 3 HEMOGLOBIN(E) procedure performed with the CAPILLARYS 3 instrument were in agreement with the reference procedure. There was no case observed of false positive, i.e., detection of an abnormal band or abnormal level of a normal band where no such abnormality existed.

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Site 2

The levels of hemoglobin fractions were measured in 151 blood samples, including 60 samples with hemoglobin variants, both by electrophoretic separations obtained with the CAPI 3 HEMOGLOBIN(E) procedure performed with the CAPILLARYS 3 instrument and a commercially available capillary electrophoresis technique for hemoglobin analysis (reference).

The measured values of hemoglobin fractions from both procedures were analyzed by three regression statistical procedures.

Normal hemoglobins

Fraction	Number ofsamples	Correlationcoefficient	Ordinary linearregression		Weighted Demingregression		Passing-Bablokregression		Range of Hb % valuesCAPI 3 HEMOGLOBIN(E)
Hb A	148	1,000	-1,928	1,020	-1,553	1,015	-1,379	1,014	15,7 - 98,3
Hb A2	151	0,987	-0,005	1,017	0,004	1,012	0,000	1,000	0,9 - 6,1
Hb F	30	0,999	-0,127	1,009	-0,021	0,965	0,000	1,000	0,5 - 34,3

Hemoglobin variants

Fraction	Number ofsamples	Correlationcoefficient	Ordinary linearregression		Weighted Demingregression		Passing-Bablokregression		Range of Hb % valuesCAPI 3 HEMOGLOBIN(E)
Hb S	33	0,999	0,475	1,006	0,105	1,015	0,272	1,011	25,8 - 78,3
Hb C	11	0,997	-1,431	1,069	-1,332	1,066	-1,480	1,067	25,2 - 37,3
Hb E	4	1,000	0,625	1,002	0,630	1,002	0,660	1,001	22,1 - 91,9

This study demonstrated a perfect correlation between the 2 analysis procedures for the Hb A, Hb A2, Hb F, Hb S, Hb C and Hb E quantitative determination.

All abnormal hemoglobins or abnormal levels of normal hemoglobins detected with the CAPI 3 HEMOGLOBIN(E) procedure performed with the CAPILLARYS 3 instrument were in agreement with the reference procedure. There was no case observed of false positive. i.e., detection of an abnormal band or abnormal level of a normal band where no such abnormality existed

The combined number of abnormal hemoglobin variants detected in the combined comparison studies is as follows: HbS=46, HbC = 24, HbD=9 HbE= 17

All abnormal hemoglobins and abnormal levels of normal hemoglobins detected were in agreement with the comparative system and clinical diagnosis. There were no observed false positives (i.e. detection of an abnormal band or abnormal level of a normal band where no such abnormal existed).

6. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Regulation Number and Section

§ 864.7415 Abnormal hemoglobin assay.

(a)
Identification. An abnormal hemoglobin assay is a device consisting of the reagents, apparatus, instrumentation, and controls necessary to isolate and identify abnormal genetically determined hemoglobin types.(b)
Classification. Class II (special controls). A control intended for use with an abnormal 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.