Who is the manufacturer of V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control?

Helena Laboratories, Corp. filed the 510(k) submission for V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control (K192931).

What is the FDA product code for V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control?

GKA. It is classified under 21 CFR 864.7415 as a Class 2 device in the Hematology panel.

What is the regulatory pathway for V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control?

510(k) clearance (submission type: Traditional).

What are the predicate devices for V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control?

Sebia CAPILLARYS Hemoglobin(E) Test (K112491).

Who can help prepare an FDA 510(k) submission like V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control

K192931 · Helena Laboratories, Corp. · GKA · Apr 19, 2022 · Hematology

Device Facts

Record ID	K192931
Device Name	V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control
Applicant	Helena Laboratories, Corp.
Product Code	GKA · Hematology
Decision Date	Apr 19, 2022
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 864.7415
Device Class	Class 2
Attributes	Pediatric

Intended Use

The V8 Nexus Hemoglobin UltraScreen method is designed for the separation of normal hemoglobins (A, A2, and F) in human blood samples, and for the detection of major hemoglobins variants (S and C) by using a capillary zone electrophoresis (CZE) buffer with the V8 instrument. The V8 Nexus Hemoglobin UltraScreen test is indicated for use in patients 2 years of age and older. This test is designed for in-vitro diagnostic use only in conjunction with other laboratory and clinical findings. The V8 instrument is an automated analyzer which performs a complete hemoglobin profile for quantitative analysis of the normal hemoglobin fractions A, A2 and F and for the detection of major hemoglobin variants S and C. The assay is performed on the hemolysate of venous whole blood collected in tubes containing K2EDTA as the anticoagulant. The V8 AFSA2 Hemo Control (Cat. No. 1812) is to be used as a quantitative and/or qualitative control for the Hemoglobin UltraScreen on the V8 Capillary Electrophoresis (CE) system.

Device Story

Device uses capillary zone electrophoresis (CZE) to separate hemoglobin fractions in human venous whole blood; input is hemolysate of K2EDTA-anticoagulated blood; system uses microbore fused silica capillary under high voltage to separate charged analytes based on differential migration; instrument performs automated sample hemolysis; output is quantitative profile of Hb A, A2, F and qualitative detection of Hb S and C; used in clinical laboratories by professionals; results used in conjunction with other clinical findings to aid in hemoglobinopathy assessment.

Clinical Evidence

No clinical data; bench testing only. Performance established via precision studies (20-day, lot-to-lot, multi-site reproducibility), linearity studies, interference testing (lipids, bilirubin, K2EDTA), and method comparison against predicate (N=439 samples). Method comparison showed high correlation (R > 0.93 for all fractions) using Passing-Bablok regression.

Technological Characteristics

Free solution capillary electrophoresis; 8 parallel capillaries; 415 nm absorbance detection; automated sample hemolysis; K2EDTA anticoagulant; liquid control material; standalone instrument platform.

Indications for Use

Indicated for patients 2 years of age and older for the separation of normal hemoglobins (A, A2, F) and detection of major variants (S, C) in venous whole blood (K2EDTA) using capillary zone electrophoresis.

Regulatory Classification

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.

Special Controls

*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.

Predicate Devices

Sebia CAPILLARYS Hemoglobin(E) Test (K112491)

Related Devices

K180762 — CAPI 3 HEMOGLOBIN(E) · Sebia · Dec 14, 2018
K112491 — SEBIA CAPILLARYS HEMOGLOBIN(E) WITH CAPILLARYS 2 · Sebia · May 24, 2012
K112550 — CAPILLARYS HEMOGLOBIN, CAPILLARYS FLEX PIERCING SYSTEM · Sebia · May 25, 2012

Submission Summary (Full Text)

{0}------------------------------------------------ Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo features the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text. April 19, 2022 Helena Laboratories, Corp. Justin Padia Director of Regulatory Affairs 1530 Lingbergh Drive Beaumont, Texas 77707 Re: K192931 Trade/Device Name: V8 Nexus Hemoglobin Ultrascreen, V8 AFSA2 Hemo Control Regulation Number: 21 CFR 864.7415 Regulation Name: Abnormal Hemoglobin Assay Regulatory Class: Class II Product Code: GKA, JBD Dated: October 16, 2019 Received: October 17, 2019 Dear Justin Padia: 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 may, 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 aware 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part {1}------------------------------------------------ 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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems. For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100). Sincerely, Min Wu. Ph.D. Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health Enclosure {2}------------------------------------------------ # Indications for Use #### 510(k) Number (if known) K192931 #### Device Name V8 Nexus Hemoglobin UltraScreen (Catalog #1828 / Catalog #801100) V8 Nexus AFSA2 Hemo Control (Catalog #1812) Indications for Use (Describe) The V8 Nexus Hemoglobin UltraScreen method is designed for the separation of normal hemoglobins (A, A2, and F) in human blood samples, and for the detection of major hemoglobins variants (S and C) by using a capillary zone electrophoresis (CZE) buffer with the V8 instrument. The V8 Nexus Hemoglobin UltraScreen test is indicated for use in patients 2 years of age and older. This test is designed for in-vitro diagnostic use only in conjunction with other laboratory and clinical findings. The V8 instrument is an automated analyzer which performs a complete hemoglobin profile for quantitative analysis of the normal hemoglobin fractions A, A2 and F and for the detection of major hemoglobin variants S and C. The assay is performed on the hemolysate of venous whole blood collected in tubes containing K2EDTA as the anticoagulant. The V8 AFSA2 Hemo Control (Cat. No. 1812) is to be used as a quantitative and/or qualitative control for the Hemoglobin UltraScreen on the V8 Capillary Electrophoresis (CE) system. | Type of Use (Select one or both, as applicable) | | |----------------------------------------------------------------------------------|--------------------------------------------------------------------------------| | <div> Prescription Use (Part 21 CFR 801 Subpart D) </div> | <div> Over-The-Counter Use (21 CFR 801 Subpart C) </div> | #### CONTINUE ON A SEPARATE PAGE IF NEEDED. This section applies only to requirements of the Paperwork Reduction Act of 1995. #### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {3}------------------------------------------------ # 510(k) Summary This section includes a description of the device: including the indications for use and technology; a comparison to the predicate device; and a concise summary for any performance testing in the submission. #### Purpose for Submission: New Device #### Type of Test: Quantitative, Capillary Zone Electrophoresis #### Submitter's Name: Helena Laboratories, Corp. | Address: | 1530 Lindbergh Drive Beaumont, TX 77707 | |----------|--------------------------------------------| |----------|--------------------------------------------| Phone: (409) 842-3714 Date Summary Prepared: 4/13/2022 #### Proprietary Names: V8 Nexus Hemoglobin UltraScreen (Catalog #1828 / Cat. #801100) V8 AFSA2 Hemo Control (Catalog #1812) #### Device Classification: #### Product: V8 Nexus Hemoglobin UltraScreen Classification: Class II Panel: Hematology Product Code: GKA C. F. R. Section: 864.7415 #### Product: V8 AFSA2 Hemo Control Classification: Class II Panel: Hematology Product Code: JBD C. F. R. Section: 864.7440 {4}------------------------------------------------ #### Intended Use: The V8 Nexus Hemoglobin UltraScreen method is designed for the separation of normal hemoglobins (A, A2, and F) in human blood samples, and for the detection of major hemoglobins variants (S and C) by using a capillary zone electrophoresis (CZE) buffer with the V8 instrument. The V8 Nexus Hemoglobin UltraScreen test is indicated for use in patients 2 years of age and older. This test is designed for in-vitro diagnostic use only in conjunction with other laboratory and clinical findings. The V8 instrument is an automated analyzer which performs a complete hemoglobin profile for quantitative analysis of the normal hemoglobin fractions A, A2 and F and for the detection of major hemoglobin variants S and C. The assay is performed on the hemolysate of venous whole blood collected in tubes containing K2EDTA as the anticoagulant. The V8 AFSA2 Hemo Control (Cat. No. 1812) is to be used as a quantitative and/or qualitative control for the Hemoglobin UltraScreen on the V8 Capillary Electrophoresis (CE) system. #### Predicate Comparison for Submission: #### Comparison with Predicate Device: These devices have the same intended use and indications for use and similar technological characteristics and principles of operation. Sample hemolysis is performed automatically by an instrument on EDTA anti-coagulant whole blood specimens. They are run on different instrumentation platforms; however, both are based on similar technological principles. The table below provides a description of Helena's device, conveying similarities and differences between the assay and the predicate device: | | Device | Predicate | |---------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | | Helena Laboratories V8 Nexus Hemoglobin UltraScreen Test | Sebia CAPILLARYS Hemoglobin(E) Test | | 510(k) Number | K192931 | K112491 | | Type of Test | Quantitative, Capillary Zone Electrophoresis | Quantitative, Capillary Electrophoresis | | Measurand | Hemoglobin A, F, A2, S, C | Hemoglobin A, F, A2, S, C, E, D | | Intended User | Clinical Laboratory Professional | Clinical Laboratory Professional | | Intended Use | In vitro diagnostic hemoglobin test for the quantitative detection of individual hemoglobin fractions A, A2 and F and qualitative detection of major hemoglobin variants S and C from EDTA anti- coagulated human whole blood specimens. | In vitro diagnostic hemoglobin test for the quantitative detection of individual hemoglobin fractions A, A2 and F and qualitative detection of major hemoglobin variants S, C, E and D from EDTA anti- coagulated human whole blood specimens. | #### V8 Nexus Hemoglobin UltraScreen Substantial Equivalence – Comparison to Predicate Device {5}------------------------------------------------ | Specimen Type | Hemolysate of venous whole blood | Hemolysate of whole blood | |---------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Anticoagulant | K2EDTA | K2EDTA and K3EDTA | | Sample Preparation | Sample hemolysis performed automatically by an instrument | Sample hemolysis performed automatically by an instrument | | Technological Detection Principles | Free solution capillary electrophoresis – protein separation occurs, in an electrolyte medium, according to their electrophoretic mobilities. Electropherograms show separated fractions based on mass to charge ratio. | Free solution capillary electrophoresis (FSCE): protein separation in an alkaline buffer (pH 9.4) according to their charge to the electrolyte pH and electroosmotic flow. Electropherograms show separated fractions according to their charge. | | Control for Migration / Other | V8 AFSA2 Hemo Control | Normal Hb A2 control; HB AFSC | | Absorbance Wavelength | 415 nm | 415 nm | | Instrument | V8 Instrument | CAPILLARYS 2 Instrument | #### Sample Hemolysis V8 Nexus Hemoglobin UltraScreen - Performed automatically by the system Sebia CAPILLARYS Hemoglobin(E) - same #### Collection Tubes V8 Nexus Hemoglobin UltraScreen - EDTA anticoagulant Sebia CAPILLARYS Hemoglobin(E) - same #### Separation System V8 Nexus Hemoglobin UltraScreen – Free solution capillary electrophoresis – protein separation occurs, in an electrolyte medium, according to their electrophoretic mobilities. Electropherograms show separated fractions based on mass to charge ratio. Sebia CAPILLARYS Hemoglobin(E) – Free solution capillary electrophoresis (FSCE): protein separation in an alkaline buffer (pH 9.4) according to their charge to the electrolyte pH and electroosmotic flow. Electrophoretograms show separated fractions according to their charge. {6}------------------------------------------------ #### Number of Separation Units V8 Nexus Hemoglobin UltraScreen - 8 parallel capillaries Sebia CAPILLARYS Hemoglobin(E) - 7 parallel capillaries #### Automated Sample Introduction V8 Nexus Hemoglobin UltraScreen - Continuous loading with sample racks Sebia CAPILLARYS Hemoglobin(E) - Continuous loading with sample racks #### Sample Processing V8 Nexus Hemoglobin UltraScreen – Aspiration of whole blood from open tube Sebia CAPILLARYS Hemoglobin(E) – Aspiration of hemolysate of packed red blood cells from uncapped tube #### Assay Throughput V8 Nexus Hemoglobin UltraScreen - 32 samples / hour Sebia CAPILLARYS Hemoglobin(E) - 33 samples/ hour #### Kit Components V8 Nexus Hemoglobin UltraScreen - All components supplied together Sebia CAPILLARYS Hemoglobin(E) - All components supplied together #### Sample Identification V8 Nexus Hemoglobin UltraScreen – Barcode reader racks and tubes Sebia CAPILLARYS Hemoglobin(E) - Barcode reader racks and tubes #### Absorbance Wavelength V8 Nexus Hemoglobin UltraScreen – 415 nm Sebia CAPILLARYS Hemoglobin(E) - 415 nm {7}------------------------------------------------ ## Controls for Test: The V8 AFSA2 Hemo Control (Cat. No. 1812) is to be used as a quantitative and/or qualitative control for the Hemoglobin UltraScreen on the V8 Capillary Electrophoresis (CE) system. ## Predicate Controls: Sebia Normal Hb A2 Control/ Migration control Sebia Hb AFSC Control ## Comparison Table | Item | Device | Predicate | |-------------------------|--------------------------------------------------------------------------------------------------------|------------------------------------------------------------------| | Product | V8 AFSA₂ Hemo Control | Normal Hb A2 Control / Migration Control & HB AFSC Control | | Intended use | Control material derived from whole blood with a quantitative range to ensure system performance | Same | | Matrix | Human serum based | Same | | Form | Liquid | Lyophilized | | Visualized fractions | Four fractions (Hb A, F, S, A₂) | Same | | Storage | 2-8°C | Same | {8}------------------------------------------------ #### Summary of Performance Testing - Bench: Performance Testing - Precision/Reproducibility Studies The precision/reproducibility of the V8 Nexus UltraScreen procedure was evaluated following CLSI EP05-A3 guidelines. The means, standard deviations and coefficients of variation were calculated for each hemoglobin fraction. #### 20-day Precision/Reproducibility Study using one instrument, one kit lot with AFSA2 and AFSC hemoglobin controls AFSA2 and AFSC hemoglobin controls were run using a single V8 Nexus instrument and a single UltraScreen kit for 20 days, twice per day with 8 reps per run. Within capillary, Between capillary, Between run, Between day and Total reproducibility estimates were calculated (mean fraction %, SD and CV). | Fraction | N | Mean % | Within Capillary SD, CV | Between Capillary SD, CV | Between Run SD, CV | Between Day SD, CV | Total SD, CV | |----------|-----|-----------|-------------------------------|--------------------------------|--------------------------|--------------------------|-----------------| | Hb A | 320 | 43.2 | 0.13, 0.3 | 1.64, 5.0 | 0.39, 1.2 | 0.39, 1.2 | 1.77, 5.3 | | Hb F | 320 | 33.1 | 0.10, 0.3 | 1.09, 3.3 | 0.26, 0.8 | 0.26, 0.8 | 1.13, 3.4 | | Hb S | 320 | 21.7 | 0.15, 0.7 | 0.09, 0.4 | 0.22, 1.0 | 0.22, 1.0 | 0.37, 1.7 | | Hb A2 | 320 | 1.9 | 0.07, 3.7 | 0.03, 1.6 | 0.07, 3.7 | 0.07, 3.7 | 0.11, 5.8 | | Fraction | N | Mean % | Within Capillary SD, CV | Between Capillary SD, CV | Between Run SD, CV | Between Day SD, CV | Total SD, CV | |----------|-----|-----------|-------------------------------|--------------------------------|--------------------------|--------------------------|-----------------| | Hb A | 320 | 29.3 | 0.09, 0.3 | 0.38, 1.3 | 0.18, 0.6 | 0.18, 0.6 | 0.47, 1.6 | | Hb F | 320 | 24.1 | 0.07, 0.3 | 0.51, 2.1 | 0.17, 0.7 | 0.19, 0.8 | 0.60, 2.5 | | Hb S | 320 | 25.0 | 0.10, 0.4 | 0.13, 0.5 | 0.15, 0.6 | 0.18, 0.7 | 0.33, 1.3 | | Hb C | 320 | 21.6 | 0.11, 0.5 | 0.04, 0.2 | 0.15, 0.7 | 0.15, 0.7 | 0.22, 1.0 | {9}------------------------------------------------ #### 5-day reproducibility study with three instruments and one kit Precision/reproducibility studies were performed using fresh and refrigerated venous K2-EDTA patient samples with known hemoglobin variants. Normal Hb AA2, Hb AF (elevated F), Hb ASA2 (elevated S and A2) and Hb AA2C (elevated C and A2) were analyzed over the course of the 5 day x 2 runs/day x 8 reps per run study. One UltraScreen lot, three V8 Nexus instruments and a single operator were employed. Within run, Between run, Between day, Between instrument and Total reproducibility estimates were calculated (mean fraction %, SD and CV). | HbA Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between instrument SD, CV | Total SD, CV | |-----------------|-----|-----------|-------------------------|--------------------------|--------------------------|---------------------------------|-----------------| | 1 | 240 | 96.9 | 0.0, 0.0 | 0.0, 0.0 | 0.10, 0.1 | 0.10, 0.1 | 0.10, 0.1 | | 2 | 240 | 34.8 | 0.35, 1.0 | 0.66, 1.9 | 0.42, 1.2 | 0.42, 1.2 | 0.87, 2.5 | | 3 | 240 | 61.0 | 0.37, 0.6 | 0.12, 0.2 | 0.43, 0.7 | 0.67, 1.1 | 0.79, 1.3 | | 4 | 240 | 58.0 | 0.35, 0.6 | 0.29, 0.5 | 0.58, 1.0 | 0.70, 1.2 | 0.93, 1.6 | | Hb F Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between instrument SD, CV | Total SD, CV | | 2 | 240 | 65.2 | 0.33, 0.5 | 0.65, 1.0 | 0.46, 0.7 | 0.39, 0.6 | 0.85, 1.3 | | Hb A2 Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between instrument SD, CV | Total SD, CV | | 1 | 240 | 3.1 | 0.05, 1.5 | 0.00, 0.1 | 0.05, 1.6 | 0.05, 1.6 | 0.07, 2.2 | | 3 | 240 | 3.6 | 0.05, 1.3 | 0.01, 0.4 | 0.05, 1.3 | 0.05, 1.3 | 0.07, 1.9 | | 4 | 240 | 5.5 | 0.07, 1.2 | 0.06, 1.1 | 0.07, 1.2 | 0.07, 1.2 | 0.12, 2.1 | | Hb S Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between instrument SD, CV | Total SD, CV | | 3 | 240 | 34.5 | 0.35, 1.0 | 0.17, 0.5 | 0.38, 1.1 | 0.59, 1.7 | 0.76, 2.2 | | Hb C Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between instrument SD, CV | Total SD, CV | | 4 | 240 | 36.5 | 0.37, 1.0 | 0.29, 0.8 | 0.58, 1.6 | 0.73, 2.0 | 0.95, 2.6 | {10}------------------------------------------------ #### Three site precision/reproducibility study using three instruments and one kit lot Site to site precision of the UltraScreen Hemoglobin assay was assessed at three different sites. Nine samples of varying levels of hemoglobin A, A2, F, S and C were prepared by controlled proportion mixing to generate hemoglobin percentages for each fraction in the normal and pathologic range, where applicable. One V8 instrument, one lot of V8 Hemoglobin UltraScreen reagent kit (same at all 3 sites (1 internal and 2 external)) and a single operator per site were utilized. The tests were run at 3 Sites x 5 days x 2 runs/day x 4 reps/run. Within run, Between run, Between day, Between site and Total reproducibility estimates were calculated (mean fraction %, SD and CV). | Hb A Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Site SD, CV | Total SD, CV | |----------------|-----|-----------|-------------------------|--------------------------|--------------------------|---------------------------|-----------------| | 1 | 120 | 97.2 | 0.00, 0.0 | 0.00, 0.0 | 0.10, 0.1 | 0.10, 0.1 | 0.10, 0.1 | | 2 | 120 | 74.7 | 0.30, 0.4 | 0.52, 0.7 | 0.37, 0.5 | 0.45, 0.6 | 0.82, 1.1 | | 3 | 120 | 87.4 | 0.26, 0.3 | 0.26, 0.3 | 0.26, 0.3 | 0.44, 0.5 | 0.61, 0.7 | | 4 | 120 | 59.3 | 0.18, 0.3 | 0.24, 0.4 | 0.18, 0.3 | 0.18, 0.3 | 0.36, 0.6 | | 5 | 120 | 76.6 | 0.23, 0.3 | 0.00, 0.0 | 0.23, 0.3 | 0.38, 0.5 | 0.54, 0.7 | | 6 | 120 | 84.9 | 0.25, 0.3 | 0.00, 0.0 | 0.34, 0.4 | 0.51, 0.6 | 0.76, 0.9 | | 7 | 120 | 57.1 | 0.34, 0.6 | 0.06, 0.1 | 0.34, 0.6 | 0.34, 0.6 | 0.51, 0.9 | | 8 | 120 | 78.9 | 0.24, 0.3 | 0.32, 0.4 | 0.32, 0.4 | 0.32, 0.4 | 0.51, 0.6 | | 9 | 120 | 87.9 | 0.18, 0.2 | 0.26, 0.3 | 0.18, 0.2 | 0.26, 0.3 | 0.44, 0.5 | | Hb F Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Site SD, CV | Total SD, CV | |-----------------|-----|-----------|-------------------------|--------------------------|--------------------------|---------------------------|-----------------| | 2 | 120 | 23.3 | 0.33, 1.4 | 0.58, 2.5 | 0.35, 1.5 | 0.42, 1.8 | 0.82, 3.5 | | 3 | 120 | 10.1 | 0.23, 2.3 | 0.25, 2.5 | 0.27, 2.7 | 0.39, 3.9 | 0.62, 6.1 | | Hb A2 Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Site SD, CV | Total SD, CV | | 1 | 120 | 2.8 | 0.03, 1.1 | 0.02, 0.7 | 0.03, 1.1 | 0.03, 1.1 | 0.05, 1.8 | | 2 | 120 | 2.0 | 0.03, 1.5 | 0.01, 0.5 | 0.03, 1.5 | 0.03, 1.5 | 0.05, 2.5 | | 3 | 120 | 2.5 | 0.05, 2.0 | 0.06, 2.4 | 0.05, 2.0 | 0.05, 2.0 | 0.09, 3.6 | | 4 | 120 | 3.5 | 0.03, 0.9 | 0.0, 0.0 | 0.03, 0.9 | 0.03, 0.9 | 0.04, 1.1 | | 5 | 120 | 3.0 | 0.04, 1.3 | 0.01, 0.4 | 0.04, 1.3 | 0.04, 1.3 | 0.06, 2.0 | | 6 | 120 | 3.2 | 0.05, 1.6 | 0.04, 1.3 | 0.05, 1.6 | 0.05, 1.6 | 0.08, 2.5 | | 7 | 120 | 5.6 | 0.06, 1.1 | 0.07, 1.3 | 0.06, 1.1 | 0.06, 1.1 | 0.11, 2.0 | | 8 | 120 | 4.0 | 0.04, 1.0 | 0.01, 0.3 | 0.04, 1.0 | 0.04, 1.0 | 0.06, 1.5 | | 9 | 120 | 3.5 | 0.05, 1.4 | 0.02, 0.6 | 0.05, 1.4 | 0.05, 1.4 | 0.07, 2.0 | | Hb S Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Site SD, CV | Total SD, CV | | 4 | 120 | 37.2 | 0.19, 0.5 | 0.26, 0.7 | 0.19, 0.5 | 0.19, 0.5 | 0.37, 1.0 | {11}------------------------------------------------ | 5 | 120 | 20.2 | 0.20, 1.0 | 0.04, 0.2 | 0.24, 1.2 | 0.36, 1.8 | 0.51, 2.5 | |----------------|-----|-----------|-------------------------|--------------------------|--------------------------|---------------------------|-----------------| | 6 | 120 | 12.1 | 0.21, 1.7 | 0.05, 0.4 | 0.31, 2.6 | 0.56, 4.6 | 0.74, 6.1 | | Hb C Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Site SD, CV | Total SD, CV | | 7 | 120 | 37.3 | 0.34, 0.9 | 0.07, 0.2 | 0.34, 0.9 | 0.37, 1.0 | 0.52, 1.4 | | 8 | 120 | 17.1 | 0.24, 1.4 | 0.29, 1.7 | 0.31, 1.8 | 0.32, 1.9 | 0.53, 3.1 | | 9 | 120 | 8.6 | 0.15, 1.7 | 0.33, 3.8 | 0.21, 2.4 | 0.24, 2.8 | 0.46, 5.3 | ## Reagent Kit Lot Precision/Reproducibility Using AFSA2 Reagent lot precision/reproducibility studies were performed using one lot of AFSA2 control, three lots of V8 Hemoglobin UltraScreen reagent kits, one V8 Nexus instrument and one operator. The tests were run 5 days x 2 runs/day x 8 reps/run. Within run, Between run, Between day, Between lot and Total reproducibility estimates were calculated (mean fraction %, SD and CV). | Sample | N | Mean % | Within run SD, CV | Between run SD, CV | Between day SD, CV | Between Lot SD, CV | Total SD, CV | |--------|-----|-----------|-------------------------|--------------------------|--------------------------|--------------------------|-----------------| | A | 240 | 42.9 | 0.26, 0.6 | 1.63, 3.8 | 0.26, 0.6 | 0.43, 1.0 | 1.72, 4.0 | | F | 240 | 33.3 | 0.17, 0.5 | 1.03, 3.1 | 0.20, 0.6 | 0.27, 0.8 | 1.10, 3.3 | | S | 240 | 21.8 | 0.26, 1.2 | 0.57, 2.6 | 0.37, 1.4 | 0.57, 2.6 | 0.92, 4.2 | | A2 | 240 | 2.0 | 0.07, 3.5 | 0.07, 3.4 | 0.09, 4.4 | 0.09, 4.5 | 0.14, 7.0 | #### Performance Testing Summary - Linearity The linearity of the V8 Nexus UltraScreen procedure was evaluated following CLSI EP06-A guidelines. Linearity studies were performed using the a single V8 Nexus Hemoglobin UltraScreen reagent kit, a single V8 Nexus and the indicated venous K2-EDTA samples containing different levels of each hemoglobin fraction (9 levels for each fraction). The tests were determined to be linear over the following ranges: HbA: 3.7-97.2%, HbF: 1.1-68.7%, HbS: 5.8-78.8%, HbA2: 1.7-7.6%, HbC: 1.4-42.6%. {12}------------------------------------------------ #### Performance Testing Summary - Limit of Detection (LOD), Limit of Quantitation (LOQ): Limit of Detection (LOD) and Limits of Quantitation (LOQ) were assigned based on the determined lower limits of linearity for each hemoglobin fraction. | Fraction | LOD/LOQ % | |----------|--------------| | Hb A | 3.7 | | Hb A2 | 1.7 | | Hb F | 1.1 | | Hb S | 5.8 | | Hb C | 1.4 | #### Performance Testing Summary - Analytical Specificity: Interference studies were conducted on venous K2-EDTA whole blood samples: 5 Hb A levels, 4 Hb A2 levels (2 normal and 2 elevated), 2 levels of Hb F, 1 level of Hb S and Hb C. Lipids (Sigma Intralipid #I141), Unconjugated Bilirubin (CalBiochem #2011-1GM) and Conjugated Bilirubin (CalBioChem #201102) and were analyzed with respect to interfering with hemoglobin quantitation. Lipids were tested at 0-25 g/L. Unconjugated and Conjugated bilirubin were tested at 0-25 mg/dL. Each hemoglobin fraction level listed above were analyzed four times per interferent level. No interference was observed. | Interferent | Maximum concentration | |------------------------|-----------------------| | Lipids | 25 g/L | | Unconjugated Bilirubin | 25 mg/dL | | Conjugated Bilirubin | 25 mg/dL | Interference: K2-EDTA Interference studies were conducted on fresh venous K2-EDTA whole blood samples: 3 Hb A levels, 3 Hb A2 levels (1 normal and 2 elevated), 1 level of Hb S and Hb C, respectively.. K2-EDTA concentrations were analyzed with respect to interfering with hemoglobin quantitation. K2-EDTA was tested at 3.6 mg/mL (normal) and 7.2 mg/mL (elevated). No interference was observed. | Interferent | Maximum concentration | |-------------|-----------------------| | K2-EDTA | 7.2 mg/mL | {13}------------------------------------------------ #### Summary of Performance Testing - Comparison Studies: Clinical testing was done at three external sites using the V8 Nexus Hemoglobin UltraScreen test to verify the performance characteristics of the system. Testing was done in accordance with the instructions specified in the V8 Operator Manual and the package insert for the assay. Three external sites compared V8 Nexus Hemoglobin UltraScreen to Sebia's Capillarys Hemoglobin(E) Test (K112491). Each site received a different lot of the V8 Nexus Hemoglobin UltraScreen reagent kits (3 in total). Fresh venous K2-EDTA-anticoagulated whole blood was used for the analysis. From a total of 439 patient samples-320 hemoglobin A, 412 hemoglobin A2, and 175 hemoglobin F quantitation's were performed. 143 presumptive hemoglobin S and 33 presumptive hemoglobin C. A single data point was collected per platform utilized. UltraScreen data points were accepted only if the data points were within the UltraScreen linearity range established for each hemoglobin fraction. Passing-Bablok regression was utilized to generate regression statistics. Site 1: The patient population for Site 1 was 40 normal samples and 141 pathologic samples were analyzed. The following individual determinations were performed: 122 hemoglobin A, 179 hemoglobin A2, 71 hemoglobin F, 70 hemoglobin S and 30 hemoglobin C. Two Hb F differences were >20%. However, these were associated with phenotypes where F is not the diagnosing variant. | Fraction | N | Range | R | Slope | 95% CI | Intercept | 95% CI | |----------|-----|-----------|-------|-------|-------------|-----------|------------| | Hb A | 122 | 8.7-97.2 | 0.999 | 1.007 | 1.000,1.017 | -0.74 | -1.67,0.20 | | Hb A2 | 179 | 1.6-6.1 | 0.995 | 1.000 | 1.000,1.000 | 0.00 | 0.00,0.00 | | Hb F | 71 | 1.1-44.3 | 0.999 | 1.000 | 0.996,1.003 | 0.10 | 0.06,0.11 | | Hb S | 70 | 19.7-76.8 | 0.998 | 0.953 | 0.941,0.965 | 2.53 | 1.89,3.04 | | Hb C | 30 | 23.6-42.3 | 0.975 | 1.00 | 0.923,1.114 | 0.85 | -2.84,3.30 | Site 2: The patient population for Site 2 was 54 normal samples and 95 pathologic samples were analyzed. The following individual determinations were performed: 119 hemoglobin A, 128 hemoglobin A2, 65 hemoglobin F, and 41 hemoglobin S. Six Hb A differences were >20%. However, these were associated with phenotypes where A is not the diagnosing variant. Five Hb A2 differences were >20%. Four were associated with hemoglobin variants where A2 is not diagnostic. The other Hb A2 difference >20% was within or below normal range for both analytes. Thirty-one Hb F differences of >20% were associated with twenty-two AFA2 phenotypes with elevated F. No change in diagnosis would result. The other nine were associated with hemoglobin phenotypes where F is not the diagnostic factor. | Fraction | N | Range | R | Slope | 95% CI | Intercept | 95% CI | |----------|-----|-----------|-------|-------|-------------|-----------|------------| | Hb A | 119 | 6.8-97.2 | 0.999 | 0.973 | 0.954,0.991 | 2.40 | 0.71,4.17 | | Hb A2 | 128 | 1.8-5.9 | 0.958 | 1.000 | 1.000,1.000 | 0.20 | 0.20,0.20 | | Hb F | 65 | 1.1-68.0 | 0.993 | 1.063 | 1.019,1.091 | 0.82 | 0.74,1.06 | | Hb S | 41 | 13.1-78.6 | 0.995 | 0.921 | 0.897,0.952 | 1.09 | -0.91,2.53 | | Hb C | ND | ND | ND | ND | ND | ND | ND | {14}------------------------------------------------ #### Site 3: The patient population for Site 3 was 38 normal samples and 71 pathologic samples were analyzed. The following individual determinations were performed: 79 hemoglobin A, 105 hemoglobin A2, 39 hemoglobin F, and 32 hemoglobin S. Three Hemoglobin C determinations were performed but not analyzed by individual site. These data points were included in the compiled analysis. Twelve Hb A2 differences were >20%. Nine were associated with hemoglobin variants where A2 is not diagnostic. Two were Hb A2 difference >20% were within or below normal range for both analytes. The other Hb A2 difference was associated with an AA2 thalassemia condition where diagnosis would not be changed. Twenty-three Hb F differences of >20% were associated with thirteen AFA2 phenotypes with elevated F. No change in diagnosis would result. The other ten were associated with hemoglobin phenotypes where F is not the diagnostic factor. | Fraction | N | Range | R | Slope | 95% CI | Intercept | 95% CI | |----------|-----|-----------|-------|-------|-------------|-----------|-----------| | Hb A | 79 | 10.5-97.2 | 0.998 | 0.973 | 0.949,0.989 | 2.55 | 1.28,4.74 | | Hb A2 | 105 | 1.9-6.9 | 0.931 | 0.963 | 0.893,1.000 | -0.09 | -2.0,0.17 | | Hb F | 39 | 1.2-31.9 | 0.988 | 1.082 | 1.000,1.149 | 0.86 | 0.57,1.10 | | Hb S | 32 | 9.6-73.9 | 0.995 | 0.908 | 0.864,0.939 | 2.37 | 1.04,3.91 | | Hb C | ND | ND | ND | ND | ND | ND | ND | Sites 1-3 Combined: | Fraction | N | Range | R | Slope | 95% CI | Intercept | 95% CI | |----------|-----|-----------|-------|-------|-------------|-----------|------------| | Hb A | 320 | 6.8-97.2 | 0.999 | 0.990 | 0.984,0.998 | 0.78 | 0.12,1.40 | | Hb A2 | 412 | 1.6-6.9 | 0.957 | 1.000 | 1.000,1.000 | 0.05 | 0.05,0.05 | | Hb F | 175 | 1.1-68.0 | 0.993 | 1.000 | 0.985,1.020 | 0.70 | 0.52,0.73 | | Hb S | 143 | 9.6-78.6 | 0.994 | 0.929 | 0.909,0.948 | 2.40 | 1.67,3.22 | | Hb C | 33 | 23.6-42.3 | 0.975 | 1.000 | 0.925,1.103 | 0.60 | -2.70,3.06 | #### Reference Range Assignment To establish a reference range, 132 normal hemoglobin samples in total were analyzed at 3 sites for Hemoglobin A and Hemoglobin A2 levels using the V8 Nexus Hemoglobin UltraScreen assay. The resulting 95% confidence limits reference ranges are as follows: #### Hb A: 96.9-97.2% Hb A2: 2.1-3.4% Note: The UltraScreen assay does not report quantitation of hemoglobin F below 0.9%. The level of Hemoglobin F in a non-neonatal population is below the reportable limit of the assay. {15}------------------------------------------------ # Conclusion: The information provided in this premarket submission is complete and has demonstrated safety and efficacy. Helena Laboratories has concluded that this submission supports a substantial equivalence decision, as compared to the predicate device.