HEMOSIL F11 & FV DNA CONTROL

K093737 · Instrumentation Laboratory CO · NZB · May 4, 2010 · Microbiology

Device Facts

• Record ID: K093737
• Device Name: HEMOSIL F11 & FV DNA CONTROL
• Applicant: Instrumentation Laboratory CO
• Product Code: NZB · Microbiology
• Decision Date: May 4, 2010
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5910
• Device Class: Class 2

Indications for Use

HemosIL FII & FV DNA Control is intended for in vitro diagnostic use as a heterozygous quality control to monitor analytical performance of the extraction, amplification and detection steps of the Xpert™ HemosIL® FII & FV genotyping assay on the GeneXpert® Dx System used in the detection of both the Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations.

Device Story

HemosIL FII & FV DNA Control is a synthetic, non-infectious quality control material containing Factor II and Factor V DNA sequences. It is formulated as a heterozygous control for Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations, suspended in a blood-like matrix. The device is used in clinical laboratory settings to monitor the analytical performance of the Xpert HemosIL FII & FV genotyping assay on the GeneXpert Dx System. Operators process the control material identically to whole blood patient samples through the system's extraction, amplification, and detection steps. The system provides qualitative genotype calls. Healthcare providers use the control results to verify the validity of patient test runs, ensuring the assay is performing correctly before reporting clinical results.

Clinical Evidence

No clinical data. Bench testing only. Performance was validated using the Xpert HemosIL Factor II & Factor V Assay on the GeneXpert Dx System. Precision/reproducibility studies (n=201 tests across multiple sites/operators) demonstrated 100% correct genotype calling. Stability studies confirmed 18-month shelf life and 30-day open-vial stability.

Technological Characteristics

Synthetic (recombinant) DNA with non-human carrier DNA, preservatives, dye, and stabilizers. Ready-to-use liquid format. Validated for use with the GeneXpert Dx System. Storage at 2-8°C. Mutation presence validated via bi-directional sequencing.

Indications for Use

Indicated for use as a heterozygous quality control for the Xpert HemosIL FII & FV genotyping assay on the GeneXpert Dx System to monitor analytical performance of extraction, amplification, and detection of Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations.

Regulatory Classification

Identification

Quality control material for cystic fibrosis nucleic acid assays. A quality control material for cystic fibrosis nucleic acid assays is a device intended to help monitor reliability of a test system by detecting analytical deviations such as those that may arise from reagent or instrument variation in genetic testing. This type of device includes recombinant, synthetic, and cell line-based DNA controls.

Special Controls

*Classification.* Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 866.9. The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: Quality Control Material for Cystic Fibrosis Nucleic Acid Assays.” See § 866.1(e) for the availability of this guidance document.

The device is classified as Class II under regulation 21 CFR 866.5910 with special controls. The special control guidance document "Class II Special Controls Guidance Document: Quality Control Material for Cystic Fibrosis Nucleic Acid Assays" is available at http://www.fda.gov/cdrh/oivd/guidance/1614.html.

Predicate Devices

• INTROL™ CF Panel I Control (K083171)

Submission Summary (Full Text)

{0}

1

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
k093737

B. Purpose for Submission:
New device

C. Measurand:
Controls for assays detecting Factor II (FII) and Factor V (FV) gene mutations

D. Type of Test:
Assayed Quality Control material

E. Applicant:
Instrumentation Laboratory Co.

F. Proprietary and Established Names:
HemosIL FII & FV DNA Control

G. Regulatory Information:
1. Regulation section:
21 CFR 866.5910; Quality control material for cystic fibrosis nucleic acid assays
2. Classification:
Class II
3. Product code:
NZB, Quality Control material, genetics, DNA
4. Panel:
Immunology (82)

H. Intended Use:
1. Intended use(s):
HemosIL FII & FV DNA Control is intended for in vitro diagnostic use as a heterozygous quality control to monitor analytical performance of the extraction, amplification and detection steps of the Xpert™ HemosIL FII & FV genotyping assay on the GeneXpert® Dx System used in the detection of both the Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations.
For in vitro diagnostic use.
2. Indication(s) for use:
Same as Intended Use
3. Special conditions for use statement(s):
For prescription use only.
4. Special instrument requirements:
GeneXpert® Dx System (k060540)

I. Device Description:
HemosIL FII & FV DNA Control is synthetic Factor II (FII) and Factor V (FV) DNA suspended in a non-infectious blood-like matrix. Each vial includes normal and mutated Factor II and Factor V DNA sequences as follows:

{1}

Table 1. No. of bases flanking each mutation site for both FII and FV

| Construct | 5' | 3' |
| --- | --- | --- |
| FII wild type | 511 | 125 |
| FII mutant | 511 | 122 |
| FV wild type | 434 | 394 |
| FV mutant | 434 | 394 |

HemosIL FII & FV DNA Control is configured as a heterozygous control for both Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations. The ratio of FII and FV DNA to non-human carrier DNA is 1:186.

The quality control material is validated for use with Xpert™ HemosIL® FII & FV genotyping assay on the GeneXpert® Dx System and is processed exactly as whole blood samples during the extraction, amplification and detection steps.

# J. Substantial Equivalence Information:

1. Predicate device name(s):

INTROL™ CF Panel I Control

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

K083171

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Physical Composition | Synthetic (recombinant) DNA with non-human carrier DNA, preservatives, dye and stabilizers | Same |
| Physical Format | Ready-to-use liquid | Same |
| Assay Steps Monitored | Extraction, Amplification and Detection | Same |
| Recommended Storage | 2-8°C | Same |
| Method to Validate Presence Mutations | Bi-directional sequencing | Same |
| Directions for Use | Handle control in the same manner as the patient sample | Same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Gene Segment | Factor II (20210G>A) Factor V (1691G>A Leiden mutation) | CFTR (38 mutations; 4 variants) |

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

None referenced.

# L. Test Principle:

Not applicable.

{2}

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

All testing of the HemosIL FII & Factor V Control was performed using the FDA cleared Xpert HemosIL Factor II & Factor V Assay (k082118) performed on the FDA cleared GeneXpert Dx System (k060540).

a. Precision/Reproducibility:

Repeatability: Two lots of HemosIL FII & FV DNA Control were tested in replicates of 5 using a single lot of Xpert HemosIL Factor II & Factor V Assay on the GeneXpert Dx instrument. In addition, five replicates of one Control lot were tested using a second lot of HemosIL Factor II & Factor V Assay. All genotypes were called correctly and results are summarized as follows:

| | | | FII G Normal | | FII A Mutant | | FV G Normal | | FV A Mutant | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Control Lot # | Assay Lot # | Correct Results (FII Het/ FV Het) | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD |
| 1 | 1 | 100% (5/5) | 27.3 | 0.4 | 26.4 | 0.5 | 27.3 | 0.6 | 27.4 | 0.7 |
| 2 | 1 | 100% (5/5) | 27.0 | 0.5 | 25.9 | 0.4 | 26.8 | 0.4 | 27.0 | 0.3 |
| 1 | 2 | 100% (5/5) | 25.9 | 0.3 | 25.4 | 0.3 | 26.1 | 0.4 | 26.2 | 0.4 |

Lot-to-Lot Variability: Three lots of HemosIL FII & FV DNA Control were tested in replicates of five using 1 lot of Xpert HemosIL Factor II & Factor V Assay on the GeneXpert Dx instrument. All genotypes were called correctly and results are summarized as follows:

| | | | FII G Normal | | FII A Mutant | | FV G Normal | | FV A Mutant | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Control Lot # | No. of Tests | Correct Results (FII Het/ FV Het) | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD |
| 1 | 22 | 100% (22/22) | 26.4 | 0.7 | 25.9 | 0.6 | 26.8 | 0.8 | 27.0 | 0.8 |
| 2 | 17 | 100% (17/17) | 26.4 | 0.6 | 25.9 | 0.6 | 26.7 | 0.7 | 27.0 | 0.7 |
| 3 | 15 | 100% (15/15) | 26.6 | 0.7 | 26.0 | 0.7 | 26.9 | 0.8 | 27.0 | 0.8 |
| ALL | 54 | 100% (54/54) | 26.4 | 0.7 | 25.9 | 0.6 | 26.8 | 0.7 | 27.0 | 0.8 |

Reproducibility: Three lots of HemosIL FII & FV DNA Control were tested using 3 different lots of Xpert HemosIL Factor II & Factor V Assay on different GeneXpert Dx instruments at multiple sites by multiple operators. A total of 201 tests were performed. All genotypes were called correctly and results are summarized as follows:

{3}

| | | | FII G Normal | | FII A Mutant | | FV G Normal | | FV A Mutant | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Control Lot # | No. of Tests | Correct Results (FII Het/ FV Het) | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD | Mean Ct | SD |
| 1 | 45 | 100% (45/45) | 26.5 | 0.9 | 25.9 | 0.8 | 26.8 | 0.9 | 27.0 | 0.9 |
| 2 | 122 | 100% (122/122) | 26.1 | 0.7 | 25.3 | 0.7 | 26.2 | 0.8 | 26.2 | 0.8 |
| 3 | 34 | 100% (34/34) | 26.8 | 0.6 | 26.0 | 0.6 | 26.9 | 0.7 | 26.9 | 0.7 |
| ALL | 201 | 100% (201/201) | 26.3 | 0.8 | 25.6 | 0.7 | 26.5 | 0.8 | 26.6 | 0.8 |

b. Linearity/assay reportable range:

Not applicable.

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

Traceability

Validation was performed by bidirectional sequencing of HemosIL FII & FV DNA Control.

Stability

Upon receipt and after opening, the material should be stored at $2^{\circ} - 8^{\circ}\mathrm{C}$ . HemosIL FII & FV DNA Control material is shipped with a "Do not freeze" warning in the device labeling.

Unopened HemosIL FII & FV DNA Control material is stable for 18 months when stored refrigerated $(2^{\circ} - 8^{\circ}\mathrm{C})$ . Opened material returned to the refrigerator $(2^{\circ} - 8^{\circ}\mathrm{C})$ shortly after use is stable for thirty (30) days from the date of opening.

Real-Time Stability Study

Three different lots of HemosIL FII & FV DNA Control were tested at defined time intervals for up to 570 days, 565 days and 64 days, respectively, using the Xpert HemosIL Factor II & Factor V Assay. Data from this ongoing real-time stability study supports a shelf-life of 18 months.

Open-Vial Stability Study

Vials from two different lots of HemosIL FII & FV DNA Control were opened, tested in duplicate at Day 0 and stored at $2 - 8^{\circ}\mathrm{C}$ for 34 days. Normal laboratory use was simulated over the test period by periodically warming the vials to room temperature and pipetting the control material, but then returning it to the vial. At Day 34, the open vials were tested in duplicate and the Ct (Crossing Threshold) results compared to the Ct results from time zero. The study demonstrated no loss of signal when used in Xpert HemosIL Factor II & Factor V Assay performed on the GeneXpert Dx Systemassay at the end of the test period.

Stress Testing Study

Shipping Study:

Two different lots of HemosIL FII & FV DNA Control were shipped in March via US Postal Service across the U.S. from Maine to California without ice packs. After receipt in California, the controls were left at room temperature for 3 days before being shipped back to Maine, again

{4}

without ice packs. The total transit time was 13 days. The returned controls were tested using Xpert HemosIL Factor II & Factor V Assay on the GeneXpert Dx System. The data demonstrates product stability during prolonged shipping without refrigeration.

Freeze/Thaw Study:
Two different lots of HemosIL FII & FV DNA Control were frozen at -20°C for 48 hours and then thawed for testing using Xpert HemosIL Factor II & Factor V Assay on the GeneXpert Dx System. The data demonstrates one freeze/thaw cycle will not degrade the product.

Expected values
The expected results with the HemosIL FII & FV DNA Control using the FDA-cleared Xpert HemosIL FII & FV genotyping assay on the GeneXpert Dx System are heterozygous genotype for both Factor II 20210G>A and Factor V 1691G>A (Leiden) mutations.

d. Detection limit:
Not applicable

e. Analytical specificity:
Not applicable

f. Assay cut-off:
Not applicable

2. Comparison studies:
a. Method comparison with predicate device:
Not applicable
b. Matrix comparison:
Not applicable

3. Clinical studies:
a. Clinical Sensitivity:
Not applicable
b. Clinical specificity:
Not applicable
c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable

4. Clinical cut-off:
Not applicable

5. Expected values/Reference range:
Not applicable

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.