RESOLVE HEMOGLOBIN KIT, JB-2 STAINING SYSTEM, MODELS FR-9120, FR-9400, FR-9360, FR-9367

K050709 · Wallac OY · GKA · Mar 8, 2006 · Hematology

Device Facts

• Record ID: K050709
• Device Name: RESOLVE HEMOGLOBIN KIT, JB-2 STAINING SYSTEM, MODELS FR-9120, FR-9400, FR-9360, FR-9367
• Applicant: Wallac OY
• Product Code: GKA · Hematology
• Decision Date: Mar 8, 2006
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7415
• Device Class: Class 2
• Attributes: Pediatric

Indications for Use

The RESOLVE® Systems Hemoglobin kit is designed to separate whole blood, cord blood or dried blood spot specimen for detection of normal and variant hemoglobins by isoelectric focusing. The kit is designed to be run on a flat-bed electrofocusing unit. This assay is intended for use as an aid in the diagnosis of neonatal and adult hemoglobinopathies.

Device Story

The RESOLVE® Hemoglobin kit uses isoelectric focusing (IEF) to separate hemoglobin variants in whole blood, cord blood, or dried blood spot specimens. Samples are applied to a precast agarose gel containing RESOLVE Ampholytes (pH 6-8). An electrical current creates a stable pH gradient; hemoglobin variants migrate until their isoelectric point (pI) matches the local pH, where they form discrete bands. Bands are visualized using the JB-2 Staining System (o-dianisidine and hydrogen peroxide), which produces an insoluble colored precipitate proportional to hemoglobin concentration. The assay is performed on a flat-bed electrofocusing unit by professional laboratory personnel. Results are interpreted visually by comparing bands to position markers, with an optional Iso Scan imaging system. The device aids in diagnosing neonatal and adult hemoglobinopathies by identifying hemoglobins F, A, E, D, S, C, G-Philadelphia, and thalassemias (e.g., Hb Bart's, HbH).

Clinical Evidence

Method comparison study conducted at three sites using 2,718 specimens (routine and retrospective DBS samples). Compared RESOLVE® Hb kit against BioRad HPLC methods. Agreement rates were 99.8% (n=850), 97.7% (n=837), and 99.1% (n=1031). Precision studies (n=216-223) showed total within-laboratory CVs of 2.8% for Hb A1c vs. Hb C and 4.4% for Hb A vs. Hb F.

Technological Characteristics

Semi-quantitative isoelectric focusing assay using agarose gel with ampholytes (pH 6-8). Requires flat-bed electrofocusing unit. Components include anode/cathode solutions, Hb elution solution, electrode wicks, and blotting materials. Visualization via JB-2 Staining System. Densitometry used for semi-quantitative analysis. Manual/semi-automated workflow.

Indications for Use

Indicated for use as an aid in the diagnosis of neonatal and adult hemoglobinopathies.

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

• BioRad VARIANT Sickle Cell Short Program (K924813)
• BioRad VARIANT nbs Sickle Cell Program (K051072)

Submission Summary (Full Text)

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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
K050709

B. Purpose for Submission:
Seek clearance for a modification to a currently cleared device (K840011)

C. Measurand:
Normal and abnormal hemoglobin

D. Type of Test:
Semi-Quantitative isoelectric focusing assay

E. Applicant:
Wallac Oy

F. Proprietary and Established Names:
Resolve Hemoglobin Kit
JB-2 Staining System

G. Regulatory Information:
1. Regulation section:
21 CFR 864.7415
2. Classification:
Class II
3. Product code:
GKA
4. Panel:

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81 Hematology

H. Intended Use:

1. Intended use(s):

The RESOLVE Systems Hemoglobin kit is designed to separate whole blood, cord blood or dried blood spot specimen for detection of normal and variant hemoglobins by isoelectric focusing.

2. Indication(s) for use:

The assay is intended for use as an aid in the diagnosis of neonatal and adult hemoglobinopathies.

3. Special conditions for use statement(s):

4. Special instrument requirements:

The kit is designed to be run on a flat-bed electrofocusing unit.

I. Device Description:

The RESOLVE Hemoglobin kit is available in 3 configurations (120 tests, 360 tests, 3600 tests). Each kit contains:

| Component | 135 Tests | 360 tests | 3600tests |
| --- | --- | --- | --- |
| Agarose IEF Gel | 5 | 5 | 50 |
| Anode Solution | 30 ml | 120 ml | 480ml |
| Cathode Solution | 30 ml | 60 ml | 240 ml |
| HB Elution Solution | 30 ml | 30 ml | 240 ml |
| IEF Electrode Wicks | 12 | 15 | 2 X 80 |
| Blotting Papers | 6 | 6 | 60 |
| Blotting Strips | 6 | - | - |
| Sample Application Templates | 5 | Order separately | Order separately |

J. Substantial Equivalence Information:

1. Predicate device name(s):

BioRad VARIANT Sickle Cell Short Program

BioRad Variant nbs Sickle Cell Program

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2. Predicate 510(k) number(s):

K924813

K051072

3. Comparison with predicate:

| Similarities | | | |
| --- | --- | --- | --- |
| Item | Device | VARIANT Sickle Cell Short Program | VARIANT nbs Sickle Cell Program |
| Intended Use | To separate whole blood, cord blood or dried blood spot specimen for detection of normal and variant hemoglobins | Qualitative screen for the presence of hemoglobin's F, A, S, D, C and E in eluates of neonatal blood collected on filter paper | Qualitative screen for the presence of hemoglobins F, A, S, D, C and E in eluates of neonatal blood collected on filter paper |
| Analytes identified | Hemoglobins F,A,E,D,S,C. In addition G and α- and β-thalassemia | Hemoglobins F,A,E,S,S and C | Hemoglobins F,A,E,D,S and C |
| Differences | | | |
| --- | --- | --- | --- |
| Item | Device | VARIANT Sickle Cell Short Program | VARIANT nbs Sickle Cell Program |
| Sample | Neonatal dried blood spots, whole blood and cord blood | Dried blood spots | Dried blood spots |
| Target Population | Neonates and adults | Neonates | Neonates |

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| Differences | | | |
| --- | --- | --- | --- |
| Item | Device | VARIANT Sickle Cell Short Program | VARIANT nbs Sickle Cell Program |
| Principle | IEF | HPLC | HPLC |

## K. Standard/Guidance Document Referenced (if applicable):

## L. Test Principle:

Whole blood or cord blood, on a filter spot or collected into an EDTA or heparinized tube, is treated with a Hb Elution Solution to inhibit methemoglobin formation. The solution is then placed on an agarose gel containing RESOLVE Ampholytes pH 6-8, and the gel is placed on the electrofocusing unit.

When an electrical current is applied to the gel, the hemoglobin variants possessing individual isoelectric points (pl's), migrate through the gel. When an individual variant's pl equals the pH in the gel, it ceases migration and forms a discrete band. When all hemoglobin bands have focused, the gel is fixed in trichloroacetic acid.

Hemoglobin bands can be visualized by using the JB-2 Staining System. Stained or unstained gels can be scanned by a densitometer to estimate the percentages of normal and variant hemoglobins.

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

Variation was estimated by measuring the distance of certain Hb bands of FASC controls used in the comparison studies. The analysis of variance was used to calculate the following:

| | n | Average distance (mm) | Within-gel precision (CV%) | Between-gel Precision (%CV) | Total precision Within laboratory (%CV) |
| --- | --- | --- | --- | --- | --- |
| HbA_{1c} vs. Hb C | 216 | 17.8 | 2.2 | 1.8 | 2.8 |

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| Hb A vs. Hb F | 223 | 2.1 | 3.3 | 2.9 | 4.4 |
| --- | --- | --- | --- | --- | --- |

b. Linearity/assay reportable range:
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
d. Detection limit:
e. Analytical specificity:
f. Assay cut-off:

# 2. Comparison studies:

a. Method comparison with predicate device:

The RESOLVE Hemoglobin system was compared to HPLC. Samples included $60\%$ from normal patients, $20\%$ from hemoglobin combinations having Hb S and $20\%$ from other hemoglobin variants to include Hb C, D, E, G-Philadelphia, $\alpha$ -thalassemia and $\beta$ -thalassemia.

| | n | HPLC | % Agreement | Lower 95% CI |
| --- | --- | --- | --- | --- |
| Site 1 | 850 | 1 | 99.8 | 99.2 |
| Site 2 | 837 | 2 | 97.7 | 96.7 |
| Site 3 | 1031 | 1 | 99.1 | 98.4 |

b. Matrix comparison:

# 3. Clinical studies:

a. Clinical Sensitivity:

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b. Clinical specificity:

c. Other clinical supportive data (when a. and b. are not applicable):

4. Clinical cut-off:

5. Expected values/Reference range:

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

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

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