HEMOCLOT QUANTI- V-L, FACTOR V-L CALIBRATOR, BIOPHEN V-L CAL (UNDILUTED), BIOPHEN ACT. PC-R CONTROL PLASMA AND BIOPHEN

K083729 · Aniara Diagnostica, LLC · GGW · May 8, 2009 · Hematology

Device Facts

• Record ID: K083729
• Device Name: HEMOCLOT QUANTI- V-L, FACTOR V-L CALIBRATOR, BIOPHEN V-L CAL (UNDILUTED), BIOPHEN ACT. PC-R CONTROL PLASMA AND BIOPHEN
• Applicant: Aniara Diagnostica, LLC
• Product Code: GGW · Hematology
• Decision Date: May 8, 2009
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7925
• Device Class: Class 2

Intended Use

HEMOCLOT Quanti VL is an in vitro diagnostic test for the quantitative determination of Factor V Leiden (FVL) activity in human citrated plasma, using automated or manual methods, by its resistance to the action of Activated Protein C ( APC). Patients with the Factor V-L (mutalion R506Q), are exposed to an increased thrombotic risk. Biophen V-L CAL (undilated), at defined Factor V-Leiden (FV-L) concentrations, for the callibration of Factor V-Leiden aclivity on human plasma, using the Hemoclot Quanti V-L kit. Factor V-L Cafibrator, pre diluted (1:20) human plasmas. at defined Factor V-Leiden (FV-L) concentrations, for the calibration of Factor V-L activity on human plasma using Hemoclot Quanti V-L Kit. Biophen Act PC-r Control Plasma contains human plasma, presenting an Activated Protein C Resistance (APC-R) is used as quality control plasma for the testing of Activated Protein C resistance (APC-R) on human plasma using Hemoclot Quarti V-L Kit. (Updated) BIOPHEN Normal Control Plasma is a set of 12 vials of normal citrated human plasma for the quality control of some coagulation factors. The following table shows parameters, which are measured using assays from HYPHEN BioMed or from other manufacturers, and according to the package inserts: Assays Reagents Manufacturers Reference ATIII Biophen ATIII Hyphen Biomed 221102/221105 Protein C Biophen Protein C Hyphen Biomed 221202/221205 aPC resistance (FV Leiden) Hemoclot Quanti V-L Hyphen Biomed CK065K Lupus Anticoagulant DVVtest®/DVVconfirm® American Diagnostica 810/815/815L DVVtest, DVVconfirm are registered trademarks from American Diagnostica, Inc. The BIOPHEN Normal Control Plasma is tested for the absence of Lupus Anticoagulant and can be used as a negative control for this investigation. This control plasma is also tested for the absence of Activated Protein C resistance (Act PC-7). When the APTT is performed with or without Activated Protein C (APC) the ratio obtained (APTT + APC/APTT) is ≥2.00.

Device Story

HEMOCLOT Quanti V-L is an in vitro diagnostic clotting assay measuring Factor V Leiden (FVL) activity in human citrated plasma. Input: patient citrated plasma. Process: plasma mixed with clotting factors (prothrombin, fibrinogen, protein S, APC) and heparin-neutralizing substance; purified Factor Xa and phospholipids added; clotting initiated by calcium. Principle: clotting time is inversely proportional to FVL concentration; resistance to APC indicates FVL mutation. Output: quantitative FVL activity level. Used in clinical laboratories; operated by technicians/lab personnel. Results interpreted by clinicians to identify APC resistance associated with Factor V R506Q mutation. Benefits: aids in diagnosis of thrombophilic conditions.

Clinical Evidence

Bench testing only. Four-site method comparison study (n=189) compared Hemoclot Quanti V-L to predicate (Coatest APCr). Results: 94.18% total agreement, 89.74% positive agreement, 97.30% negative agreement. Comparison with molecular biology diagnosis (n=63) showed 97% agreement. Precision studies (intra- and inter-assay) yielded CV < 6%. Linearity validated between 10% and 100% FVL (r² > 0.98). Specificity confirmed with factor-deficient plasmas and samples with heparin or AVK therapy.

Technological Characteristics

Lyophilized reagents (human fibrinogen, prothrombin, Protein S, APC, Factor Xa, rabbit brain phospholipids). Clotting-based assay. Quantitative measurement via calibration curves. Compatible with automated coagulation analyzers (e.g., STAR, KC10) or manual water bath. Reagents stable at 2-8°C.

Indications for Use

Indicated for the quantitative measurement of Factor V Leiden activity in human citrated plasma to assess resistance to Activated Protein C (APC) in patients suspected of having coagulation disorders. For prescription use only.

Regulatory Classification

Identification

A partial thromboplastin time test is a device used for primary screening for coagulation abnormalities, for evaluation of the effect of therapy on procoagulant disorders, and as an assay for coagulation factor deficiencies of the intrinsic coagulation pathway.

Predicate Devices

• COATEST APC Resistance V (K963111)

Submission Summary (Full Text)

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— 8 2009

# Premarket Notification 510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

| The assigned 510(k) number is: | K08 3729 |
|-----------------------------------------|----------------------------------------------------------------------------------------------------------------------------|
| Submitted by: | Aniara Diagnostica, LLC<br>6560 Gove Ct<br>Mason, Ohio 45040 |
| Contact Person: | Mr. Ola Andersson |
| Phone Number: | [phone] |
| Fax Number: | [phone] |
| Email address: | [e-mail] |
| Date of 510(k) Preparation: | December, 2008 |
| Name of the Devices: | 1. HEMOCLOT Quanti V-L<br>2. Factor V-L Calibrator<br>3. BIOPHEN V-L CAL (Undiluted)<br>4. BIOPHEN Act PC-r Control Plasma |
| Classification Name: | Test, Partial Thrombin Time<br>Secondary Calibrator<br>Coagulation Control Plasma |
| Class: | II |
| Classification Panels: | Hematology (81) |
| Product Code: | GGW, Test, Partial Thrombin Time<br>JIT, Calibrator, Secondary<br>GGN, Plasma, Coagulation Control |
| Regulation numbers: | 864.7925<br>862.1150<br>864.5425 |
| Identification of the Predicate Device: | |

COATEST APC Resistance V (K963111) (Chromogenix)

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#### Description of the Device:

[All devices are manufactured by Hyphen-Biomed]

- 1. HEMOCLOT Quanti V-L is an in vitro diagnostic test kit containing 3 vials of each of the following 2 lyophilized reagents:
R1: Reagent 1: Clotting mixture containing human Fibrinogen, human prothrombin, Protein S at a constant concentration, optimized for the assay, and human Activated Protein C, lyophilized. It also contains a heparin neutralizing substance. Each vial is to be reconstituted with exactly 2 ml of distilled water.

R2: Reagent 2: Purified Human Factor Xa, containing rabbit brain phospholipds (cephalin), lyophilized. Each vial is to be reconstituted with exactly 1 ml of distilled water.

Calibrators and controls to be used with the test kit are supplied separately. The following calibrators and control are included in this submission:

- 2. Factor V-L Calibrator is a calibrator kit containing 3 vials each of 1.0 ml prediluted human plasma (1:20), freeze dried, at 4 different concentrations of FV-L, to cover the assay range, from 10% to 100%. Each vial is restored with 1.0 ml Owren Koller type buffer.
- 3. BIOPHEN V-L CAL (Undiluted) is a calibrator kit containing 3 vials each of 0.5 ml undiluted human plasma, freeze dried, at 3 different concentrations of FV-L, to cover the assay range, from about 10% to 100%. Each vial is restored with 0.5 ml distilled water.
- BIOPHEN Act PC-r Control Plasma is a controls kit containing 12 vials of 0.5 ml human 4. plasma, presenting an Activated Protein C Resistance (APC-R), citrated and Iyophilized. Each vial is to be reconstituted with exactly 0.5 ml of distilled water.
- BIOPHEN Normal Control Plasma manufactured by Hyphen-Biomed, is also used with the ട് test kit and is currently marketed in the US under 510(k) # K043451, It is a controls kit containing 12 vials of 1.0 ml of normal citrated human plasma, lyophilized. Each vial is to be reconstituted with exactly 1.0 ml of distilled water.

#### Intended Use:

- 1. HEMOCLOT Quanti V-L is a clotling method for measuring the Factor V Leiden (FVL) activity in human citrated plasma, by its resistance to the action of Activated Protein C (APC). The assay is performed in the presence of Activated Protein S (one single test for each patient). In the presence of APC, the prolongation of clotting time is an inverse relationship of the concentration of Factor V-Leiden (mutation R506Q). Normal Factor V is not measured.
- 2. Factor V-L Calibrator is Iyophilized, pre-diluted (1:20) human plasmas, at defined Factor V-Leiden (FV-L) concentrations, for the calibration of Factor V-L activity quantitative clotting assay on human citrated plasma, using the HEMOCLOT Quanti. V-L Kit (ref CK065K).
- BIOPHEN V-L CAL (Undiluted) is Iyophilized, undiluted human plasmas, at defined Factor V-3. Leiden (FV-L) concentrations for the calibration curve of Factor V-L activity quantitative clotting assay on human citrated plasma, using HEMOCLOT Quanti V-L Kit (ref CK065K).
- 4. BIOPHEN Act PC-r Control Plasma kit contains human plasma, presenting an activated Protein C Resistance (APC-R), usually correlated with the genetic mutation of Factor V

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R506Q. This plasma is used as quality control plasma for the testing of Activated Protein C Resistance (APC-R).

- 5. (Updated) BIOPHEN Normal Control Plasma is a set of 12 vials of normal citrated human plasma for the quality control of some coagulation factors. The following table shows the various parameters, which are measured using assays from HYPHEN BioMed or from other manufacturers, and according to the package inserts:

| Assays | Reagents | Manufacturers | Reference |
|----------------------------|----------------------|-------------------------|---------------|
| ATIII | Biophen ATIII | Hyphen Biomed | 221102/221105 |
| Protein C | Biophen Protein C | Hyphen Biomed | 221202/221205 |
| aPC resistance (FV Leiden) | Hemoclot Quanti V-L | Hyphen Biomed | CK065K |
| Lupus<br>Anticoagulant | DVVtest®/DVVconfirm® | American<br>Diagnostica | 810/815/815L |

DVVtest, DVVconfirm are registered trade marks from American Diagnostica, Inc.

The BIOPHEN Normal Control Plasma is tested for the absence of Lupus Anticoagulant and can be used as a negative control for this investigation. This control plasma is also tested for the absence of Activated Protein C resistance (Act PC-r). When the APTT is performed with or without Activated Protein C (APC) the ratio obtained (APTT + APC/APTT) is ≥ 2.00.

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#### Statement of Technological Characteristics of the Device Compared to Predicate Device:

HEMOCLOT Quanti V-L, BIOPHEN V-L CAL (Undiluted), Factor V-L Calibrator, BIOPHEN Act PC-r Control Plasma, and BIOPHEN Normal Control Plasma are substantially equivalent in performance, intended use and safety and effectiveness to COATEST APC Resistance V (K963111).

| Characteristics | HEMOCLOT Quanti V-L, Factor<br>V-L Calibrator, Biophen V-L<br>CAL (undiluted), Biophen Act<br>PC-r Control Plasma, Biophen<br>Normal Control Plasma | COATEST APC Resistance V<br>(K963111). Control Plasmas Level 1<br>and Level 2 are included in this<br>device. |
|------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Similarities | | |
| Intended Use | Measurement of Factor V Leiden<br>activity in human citrated plasma<br>by its resistance to the action of<br>Activated Protein C. | Same |
| Assay Type | Clotting Method | Same |
| Test Sample | Human Citrated Plasma | Same |
| Assay Principle | Measurement of Factor V Leiden<br>coagulation activity based on<br>insensitivity of Factor V Leiden to<br>the action of Activated Protein C. | Same |
| Stability of<br>Unopened<br>Reagents | Stable at 2-8°C until expiry date | Same |
| Controls,<br>Normal | Citrated human plasma,<br>lyophilized, to provide quality<br>control in the normal range. APC V<br>Ratio > 2 | Same |
| Controls,<br>Abnormal (APC<br>Resistant) | Citrated human plasma,<br>lyophilized, to provide quality<br>control for APC resistant range of<br>the assay. APC V Ratio < 1.80 | Same. |
| Differences | | |
| Assay Principle | Assay is performed in the<br>presence of Activated Protein C<br>and Protein S (one single test for<br>each patient). In the presence of<br>APC, prolongation of clotting time<br>is inversely related to the amount<br>of Factor V Leiden. Normal Factor<br>V is not measured. APC<br>resistance due to Factor V Leiden<br>is indicated when the %FVL value<br>is above or equal to the cut-off<br>value. | Assay is performed in the presence<br>and absence of Activated Protein C<br>(two tests for each patient). In the<br>presence of APC, prolongation of<br>clotting time is directly related to the<br>concentration of Normal Factor V, and<br>inversely related to the amount of<br>Factor V Leiden. APC resistance duethe APC-V ratio is below or equal to<br>the cut-off value. |
| Assay<br>Calibration | Assay of calibrator plasma at<br>defined Factor V Leiden | Not Applicable |
| | | |
| | available in prediluted (FVL<br>Calibrator) and undiluted<br>(BIOPHEN V-L Cal) form,<br>lyophilized. | |
| Cut-Off Values | Normal plasmas measure ≤10%<br>FVL. APCr plasmas measure<br>>25% FVL. | Normal cut-off value (ratio) determined<br>by assay of normal patient plasmas<br>and statistical analysis of the results. |
| Test Sample<br>Dilution | Test sample diluted in Owren<br>Koller type buffer (not included in<br>kit). | Test sample diluted with V-Def Plasma<br>(included in kit). |
| Controls,<br>Normal | Biophen Normal Control Plasma<br>sold separately from kit. | Control Plasma Level 1 included in kit. |
| Controls,<br>Abnormal (APC<br>Resistant) | Biophen Act. PC-r Control Plasma<br>sold separately from kit. | Control Plasma Level 2 included in kit. |
| Controls,<br>Normal &<br>Abnormal (APC<br>Resistant) | %FVL < 10% (Normal Control);<br>%FVL between 25% and 75%<br>(APCr Control) | Value of APC-R provided as ratio only. |
| Stability of<br>Reconstituted<br>Reagents | R1 and R2 reagents: 24 hrs at 2-8°C;<br>12 hrs at 18-25°C; 1 month<br>frozen at -20°C or below.<br>Control Plasmas and Calibrators:<br>8 hrs at room temperature; 24h at<br>2-8°C; Do Not Freeze. | When stored in original vials: V-Def<br>Plasma: 8 hrs at 15-25°C; 24 hrs at 2-<br>8°C; 3 months at -20°C or below.<br>CaCl₂ and APTT reagent: 1 month at<br>2-8°C; 1 week at 15-25°C. APC/CaCl₂<br>reagent: 2 hrs at 37°C; 8 hrs at 15-25°<br>C; 5 days at 2-8°C; 3 months at -20°C<br>or below. Control Plasmas: 6 hrs at 2-<br>25°C or 3 months at -20°C or below. |
| | | |

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Aniara Diagnostica, LLC
Premarket Notification Section 510(k)
Hemoclot Quanti V-L; Factor V-L Calibrator: Biophen V-L Calibrator: Biophen V-L Calibration Actives: A Controlle

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#### Summary of Performance Data:

#### Hemoclot Quanti V-L

The HEMOCLOT Quanti V-L (Hyphen-Biomed) assay shows good consistency with Coatest APCr kit (Chromogenix), as shown from data combined from 4 studies (2 in US, 2 in Europe):

| All Sites | | Coatest APCr | |
|------------------------|---------------|--------------|----------|
| | | Normal | Abnormal |
| Hemoclot<br>Quanti V-L | Normal | 108 | 7 |
| | Abnormal | 1 | 70 |
| | Inconclusive* | 2 | 1 |
| Agreement | | 94.18% | |
| Co-positivity | | 97.30% | |
| Co-negativity | | 89.74% | |
| Sample Size | | 189 | |

*Outside of the range of the assay, between 10% and 25% FVL.

The Hemoclot Quanti VL Assay shows good consistency with Molecular Biology results:

| | | Molecular<br>Biology | |
|---------------|----------|----------------------|-----------|
| | | Normal | Abnormal. |
| Hemoclot | Normal | 15 | 0 |
| FVL-Q | Abnormal | 0 | 6 |
| Agreement | | 100.00% | |
| Co-positivity | | 100.00% | |
| Co-negativity | | 100.00% | |
| Sample Size | | 21 | |

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Reproducibility of the Hemoclot Quanti V-L assay for 2 plasmas at different FVL concentrations, using the KC10, STAR or Water Bath instrument:

| Sample | Intra Assay CV% | | | Inter Assay CV% | | |
|---------------------|-----------------|------|----|-----------------|------|----|
| | CT (sec) | %FVL | N | CT (sec) | %FVL | N |
| Sample 1 (100% FVL) | 4.4% | 5.9% | 10 | 2.2% | 3.3% | 10 |
| Sample 2 (50% FVL) | 4.2% | 8.2% | 10 | 2.3% | 4.7% | 10 |

| Instrument | Sample | Inter Assay CV% | | |
|------------|--------------------|-----------------|---------|----|
| | | On CT (sec) | On %FVL | N |
| KC10 | Sample 1 (25% FVL) | 7.3% | 17.4% | 5 |
| | Sample 2 (10% FVL) | 6% | 29.1% | 5 |
| STAR | Sample 1 (25% FVL) | 5.2% | 13.1% | 10 |
| | Sample 2 (10% FVL) | 5.6% | 23.7% | 10 |
| WATER BATH | Sample 1 (25% FVL) | 2.7% | 10.0% | 5 |
| | Sample 2 (10% FVL) | 5.6% | 15.2% | 5 |

#### Conclusion:

The performance studies demonstrate that the device is substantially equivalent to the predicate device in safety and effectiveness.

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## DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/7/Picture/1 description: The image shows the logo for the Department of Health & Human Services (USA). The logo features a stylized eagle with three stripes representing the department's mission. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - (USA)" is arranged in a circular pattern around the eagle.

#### Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

MAY -8 2009

Aniara Diagnostica, LLC c/o Ms. Ola Anderson 6560 Gove Ct. Mason, OH 45040

Re: k083729

Trade/Device Name: HEMOCLOT Quanti V-L, Factor V-L Calibrator, Biophen V-L Cal (Undiluted), Biophen ACT PC-r Control Plasma and Biophen Normal Control

Regulation Number: 21 CFR 864.7925 Regulation Name: Partial thromboplastin time tests Regulatory Class: Class II Product Code: GGW, GGN, JIT Dated: April 07, 2009 Received: April 20, 2009

Dear Ms. 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 may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice

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### Page 2 - Ms. Ola Anderson

requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at [phone]. For questions regarding the reporting of device adverse events (Medical Device Reporting (MDR)), please contact the Division of Surveillance Systems at [phone]. For more information regarding the reporting of adverse events, please go to http://www.fda.gov/cdrh/mdr/.

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

m. Chen

Maria M. Chan, PhD Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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K083729

#### Indications for Use

HEMOCLOT Quanti VL is an in vitro diagnostic test for the quantitative determination of Factor V Leiden (FVL) activity in human citrated plasma, using automated or manual methods, by its resistance to the action of Activated Protein C ( APC).

Patients with the Factor V-L (mutalion R506Q), are exposed to an increased thrombotic risk.

Biophen V-L CAL (undilated), at defined Factor V-Leiden (FV-L) concentrations, for the callibration of Factor V-Leiden aclivity on human plasma, using the Hemoclot Quanti V-L kit.

Factor V-L Cafibrator, pre diluted (1:20) human plasmas. at defined Factor V-Leiden (FV-L) concentrations, for the calibration of Factor V-L activity on human plasma using Hemoclot Quanti V-L Kit.

Biophen Act PC-r Control Plasma contains human plasma, presenting an Activated Protein C Resistance (APC-R) is used as quality control plasma for the testing of Activated Protein C resistance (APC-R) on human plasma using Hemoclot Quarti V-L Kit.

(Updated) BIOPHEN Normal Control Plasma is a set of 12 vials of normal citrated human plasma for the quality control of some coagulation factors. The following table shows parameters, which are measured using assays from HYPHEN BioMed or from other manufacturers, and according to the package inserts:

| Assays | Reagents | Manufacturers | Reference |
|-------------------------------|----------------------|-------------------------|---------------|
| ATIII | Biophen ATIII | Hyphen Biomed | 221102/221105 |
| Protein C | Biophen Protein C | Hyphen Biomed | 221202/221205 |
| aPC resistance<br>(FV Leiden) | Hemoclot Quanti V-L | Hyphen Biomed | CK065K |
| Lupus<br>Anticoagulant | DVVtest®/DVVconfirm® | American<br>Diagnostica | 810/815/815L |

DVVtest, DVVconfirm are registered trademarks from American Diagnostica, Inc.

The BIOPHEN Normal Control Plasma is tested for the absence of Lupus Anticoagulant and can be used as a negative control for this investigation. This control plasma is also tested for the absence of Activated Protein C resistance (Act PC-7). When the APTT is performed with or without Activated Protein C (APC) the ratio obtained (APTT + APC/APTT) is ≥2.00.

Demellax

**Division Sign-Off**

Office of In Vitro Diagnostic
Device Evaluation and Safety

510(k) K083729