ZYMUTEST HIA IGGAM, ZYMUTEST IGG, MODEL RK040D, RK040A

K071255 · Hyphen Biomed · LCO · Sep 7, 2007 · Hematology

Device Facts

• Record ID: K071255
• Device Name: ZYMUTEST HIA IGGAM, ZYMUTEST IGG, MODEL RK040D, RK040A
• Applicant: Hyphen Biomed
• Product Code: LCO · Hematology
• Decision Date: Sep 7, 2007
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7695
• Device Class: Class 2

Indications for Use

ZYMUTEST HIA IgG and IgGAM kits are designed as a solid phase enzyme- linked immunosorbent assay (ELISA). These products are intended to be used as an in vitro diagnostics kit by Hematology, coagulation or other pathology laboratories to assist in screening patients samples for the presence of heparin- associated antibodies commonly found in patients with heparin induced thrombocytopenia or thrombosis (HIT).

Device Story

ZYMUTEST HIA is a solid-phase ELISA kit for detecting anti-heparin/Platelet Factor 4 (PF4) antibodies in human citrated plasma or serum. Used in clinical laboratories by technicians or pathologists to assist in diagnosing heparin-induced thrombocytopenia (HIT) or thrombosis. The assay utilizes a sandwich ELISA technique; patient samples are incubated in wells coated with PF4/heparin complexes; bound antibodies are detected via enzyme-linked secondary antibodies. Results are interpreted by laboratory personnel to aid clinical decision-making regarding heparin therapy management. Benefits include identifying patients at risk for or experiencing HIT, allowing for timely clinical intervention.

Clinical Evidence

Performance evaluated via method comparison studies against predicate devices (Asserachrom, GTI PF4 Enhanced) and SRA. Studies involved 174 to 345 samples. ZYMUTEST IgGAM vs. Asserachrom showed 76% agreement (64% co-positivity, 81% co-negativity). ZYMUTEST IgGAM vs. GTI PF4 Enhanced showed 74% agreement (58% co-positivity, 90% co-negativity). ZYMUTEST IgG vs. Asserachrom showed 76% agreement (44% co-positivity, 90% co-negativity). ZYMUTEST IgG vs. SRA showed 75.3% matching. Precision testing (intra/inter-assay) showed CVs ranging from 3.07% to 7.50%.

Technological Characteristics

Solid-phase enzyme-linked immunosorbent assay (ELISA) using a sandwich technique. Analyte: anti-heparin/PF4 antibodies. Sample type: citrated human plasma or serum. Connectivity: None (manual/standalone laboratory assay). Sterilization: Not applicable (in vitro diagnostic reagent).

Indications for Use

Indicated for use in hematology, coagulation, or pathology laboratories to screen patient samples for heparin-associated antibodies in patients suspected of having heparin-induced thrombocytopenia (HIT) or thrombosis.

Regulatory Classification

Identification

A platelet factor 4 radioimmunoassay is a device used to measure the level of platelet factor 4, a protein released during platelet activation by radioimmunoassay. This device measures platelet activiation, which may indicate a coagulation disorder, such as myocardial infarction or coronary artery disease.

Predicate Devices

• DIAGNOSTICA STAGO ASSERCHROM® HPIA TEST KIT (K003767)
• GTI PF4 ENHANCED SOLID PHASE ELISA (K053559)

Submission Summary (Full Text)

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

A. 510(k) Number:
K071255

B. Purpose for Submission:
clearance of a new device

C. Measurand:
IgG, IgA, and IgM heparin-dependent antibodies

D. Type of Test:
Enzyme linked immunosorbent assay

E. Applicant:
Hyphen BioMed

F. Proprietary and Established Names:
ZYMUTEST HIA IgG
ZYMUTEST HIA IgGAM

G. Regulatory Information:
1. Regulation section:
21 CFR 864.7695

2. Classification:
Class II

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3. Product code:
LCO

4. Panel:
81 Hematology

H. Intended Use:

1. Intended use(s):
The ZYMUTEST HIA IgGAM ELISA is a qualitative screening assay intended for the global detection of heparin-dependent antibodies, whether the isotype is: IgG IgM, and IgA, in human plasma, by clinical laboratories. It is intended for in vitro diagnostic use.

The ZYMUTEST HIA IgG ELISA is a qualitative assay intended for the detection of heparin-dependent antibodies of the IgG isotope, in human plasma, by clinical laboratories. It is intended for in vitro diagnostic use.

2. Indication(s) for use:
ZYMYUTEST HIA IgG AND IgGAM KITS are designed as a solid phase enzyme-linked immunosorbent assay (ELISA). These products are intended to be used as an in vitro diagnostics kit by Hematology, coagulation or other pathology laboratories to assist in screening patient samples for the presence of heparin-associated antibodies commonly found in patients with heparin induced thrombocytopenia or thrombosis (HIT).

3. Special conditions for use statement(s):

4. Special instrument requirements:

I. Device Description:
The assay kit consists of a 96 welled micro ELISA plate coated with unfractionated heparin, 3 vials each of lyophilized Positive and Negative controls, 3 vials lyophilized Platelet lysate, 3 vials of Immunoconjugate, 25 mL of TMB substrate, 1 vial of sulfuric Acid Stop solution, buffer, diluents, and wash solution.

J. Substantial Equivalence Information:

1. Predicate device name(s):

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1. DIAGNOSTICA STAGO ASSERCHROM® HPIA TEST KIT
2. GTI PF4 ENHANCED SOLID PHASE ELISA

2. Predicate 510(k) number(s):
1. K003767
2. K053559

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Intended Use | A qualitative in vitro diagnostic screening assay intended for the global detection of IgG, IgM, and IgA heparin-dependent antibodies in human plasma, by clinical laboratories. | same |
| test principle | ELISA | same |
| sample requirements | citrated plasma | same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| materials | Microtiter plate coated with unfractionated heparin | microtiter plate coated with purified PF4 complexed to polyvinly sulfonate |

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

L. Test Principle:
When a diluted plasma sample and platelet lysate is added to one of the microwells of the coated plate, if present, heparin-dependent antibodies present in the sample will form complexes with unfractionated heparin immobilized on the plate. Following a washing step, bound antibodies are mixed with the immunoconjuuagate, which will bind to IgG, IgM, and IgA isotypes. Following another washing step, the TMB peroxidase substrate is added, and a blue color develops which turns yellow upon the addition of the stop

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solution. The color that develops is directly proportional to the amount of heparin-dependent antibodies present in the tested sample.

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

### 1. Analytical performance:

#### a. Precision/Reproducibility:

Intra-assay- ZYMUTEST HIA IgG and ZYMUTEST HIA IgGAM were tested in duplicate using positive control material.

| Positive control | N | Mean A450 | CV% |
| --- | --- | --- | --- |
| Anti PF4-IgG Lot 061027D | 6 | 1.31 | 3.07 |
| Anti PF4-IgG Lot 061214A | 9 | 1.10 | 4.46 |
| Anti PF4-IgGAM Lot 061214D | 9 | 1.74 | 4.75 |

Inter-assay-

| Positive control | N | Mean A450 | CV% |
| --- | --- | --- | --- |
| Anti PF4-IgG Lot 061027D | 7 | 1.34 | 7.11 |
| Anti PF4-IgGAM Lot 061027G | 7 | 1.84 | 7.50 |

#### b. Linearity/assay reportable range:

n/a

#### c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Negative controls are derived from normal human plasma with an A450 of mean ± 2 SD.

The positive control is derived from chimeras made by coupling anti-PF4 polyclonal antibodies with human Igs (IgG, IgA or IGM, or the mixture of the three), with an A450 of mean ± 5 SD.

#### d. Detection limit:

n/a

#### e. Analytical specificity:

The effect of varying concentrations of heparin in the assay was tested on 3 pathological plasmas with and without heparin addition,

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ranging from 0 to 5 IU/ml. After heparin addition, plasma was diluted 1:100 and tested. Results demonstrated no heparin interference up to 1 IU/ml.

f. Assay cut-off:

The assay was evaluated in hospital patients with no HIT, ACC patients with circulating anticoagulant antibodies with no HIT, and normal healthy donors with no autoimmunity background. Cut-off was established as mean + 3SD for the normal hospital patients and ACC patients, which corresponded to + 5 SD for normal healthy donors.

2. Comparison studies:

a. Method comparison with predicate device:

ZYMUTEST IgGAM compared with Asserachrom. N= 44.

| In-house Study | | Asserachrom | |
| --- | --- | --- | --- |
| | | Positive | Negative |
| Zymutest | Positive | 28 | 2 |
| IgGAM | Negative | 0 | 14 |
| Agreement | | 100% | 88% |

2 site clinical study ZYMUTEST IgGAM compared with Asserachrom

| Combined | | Asserachrom | |
| --- | --- | --- | --- |
| Site 1 & 2 | | Positive | Negative |
| Zymutest | Positive | 48 | 32 |
| IgGAM | Negative | 27 | 136 |
| Agreement | | 76% | |
| Co-positivity | | 64% | |
| Co-negativity | | 81% | |
| Sample Size | | 243 | |

3 site clinical study ZYMUTEST IgGAM compared with GTI PF4 Enhanced

| Combined | | GTI PF4-Enhanced | |
| --- | --- | --- | --- |
| Site 1, 2, &3 | | Positive | Negative |
| Zymutest | Positive | 101 | 17 |
| IgGAM | Negative | 74 | 153 |
| Agreement | | 74% | |
| Co-positivity | | 58% | |
| Co-negativity | | 90% | |
| Sample Size | | 345 | |

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2 site clinical study- ZYMUTEST IgG compared with Asserachrom

| Combined | | Asserachrom | |
| --- | --- | --- | --- |
| Site 1 & 2 | | Positive | Negative |
| Zymutest IgG | Positive | 33 | 17 |
| | Negative | 42 | 151 |
| Agreement | | 76% | |
| Co-positivity | | 44% | |
| Co-negativity | | 90% | |
| Sample Size | | 243 | |

ZYMUTEST IgG compared with Serotonin Release Assay (SRA) n=174

| # Matches | 131 |
| --- | --- |
| % Matching | 75.3% |

b. Matrix comparison:

n/a

3. Clinical studies:

a. Clinical Sensitivity:

n/a

b. Clinical specificity:

n/a

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

n/a

4. Clinical cut-off:

n/a

5. Expected values/Reference range:

n/a

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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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