INTROL CF PANEL I CONTROL

DEN060007 · Maine Molecular Quality Controls, Inc. · NZB · Oct 12, 2006 · Microbiology

Device Facts

• Record ID: DEN060007
• Device Name: INTROL CF PANEL I CONTROL
• Applicant: Maine Molecular Quality Controls, Inc.
• Product Code: NZB · Microbiology
• Decision Date: Oct 12, 2006
• Decision: DENG
• Submission Type: Post-NSE
• Regulation: 21 CFR 866.5910
• Device Class: Class 2

Indications for Use

INTROL™ CF Panel I Control is intended for in vitro diagnostic use as a quality control to monitor analytical performance of the extraction, amplification and detection steps of diagnostic assays used in the detection of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene mutations and variants. This product is intended to be extracted and analyzed routinely with each CFTR assay run.

Device Story

INTROL™ CF Panel I Control consists of synthetic CFTR DNA suspended in a matrix of synthetic DNA targets, carrier DNA, preservatives, dye, and stabilizers. The device mimics a human whole blood specimen; it is processed through standard clinical laboratory extraction methods. Following extraction, the released synthetic DNA is used in molecular amplification-based assays to monitor the performance of the extraction, amplification, and detection steps. The control contains 38 specific CFTR mutations, polymorphisms, and variants. It is used by laboratory technicians in clinical settings to ensure assay reliability. Healthcare providers use the resulting quality control data to validate that the diagnostic assay is functioning correctly for each run, thereby ensuring the accuracy of patient test results.

Clinical Evidence

Bench testing only. Performance evaluated at 10 external sites (8 clinical laboratories) using 11 manufacturing lots across 66 runs. Testing demonstrated 100% correct calls (1133/1133) across multiple amplification methods. Stability studies (real-time, stress, freeze/thaw, open vial) confirmed product integrity and signal consistency over 12 months and under various environmental conditions.

Technological Characteristics

Synthetic CFTR DNA in a matrix of synthetic DNA targets, non-human carrier DNA, preservatives, dye, and stabilizers. Contains 27 CFTR gene exons plus intronic borders. Form factor: 5 liquid bottles (versions G106ac or G106de). Storage: 2-8°C. No energy source. Not software-based.

Indications for Use

Indicated for use as a quality control material to monitor the analytical performance of extraction, amplification, and detection steps in diagnostic assays for 38 CFTR gene mutations and variants, including those recommended by ACMG and ACOG, in patients undergoing cystic fibrosis genetic testing.

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.

Reference Devices

• Tag-It™ Cystic Fibrosis (CF) Kit

Submission Summary (Full Text)

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### 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

- A. 510(k) Number: k060070
- B. Purpose for Submission: New device
- C. Measurand:

Controls for assays detecting Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene mutations and variants

- D. Type of Test: Assayed quality control material
- E. Applicant: Maine Molecular Quality Controls, Inc. (MMQCI)
- F. Proprietary and Established Names: INTROL™ CF Panel I Control
- G. Regulatory Information:
- 1. Regulation section: 866.5910 DNA quality control material for genetic testing
- 2. Classification: De novo, Class II
- 3. Product code: NZB, Quality control material, genetics, DNA
- 4. Panel: Immunology (82)

# H. Intended Use:

- 1. Intended use(s):
INTROL™ CF Panel I Control is intended for in vitro diagnostic use as a quality control to monitor analytical performance of the extraction, amplification and detection steps of diagnostic assays used in the detection of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene mutations and variants. This product is intended to be extracted and analyzed routinely with each CFTR assay run.

- 2. Indication(s) for use:
INTROL™ CF Panel I Control is intended for in vitro diagnostic use as a quality control to monitor analytical performance of the extraction, amplification and detection steps of diagnostic assays used in the detection of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene mutations and variants. This product is intended to be extracted and analyzed routinely with each CFTR assay run.

The INTROL™ CF Panel I Control is designed to monitor the detection of 38 CFTR mutations associated with cystic fibrosis, including the 23 mutations recommended for testing by American College of Medical Genetics (ACMG) and American College of Obstetricians and Gynecologists (ACOG). The INTROLTM CF Panel I Control also monitors 3 polymorphisms (1506V, 1507V, F508C) and

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the 5/7/9T variants.

- 3. Special conditions for use statement(s): For prescription use only.
The INTROL™ CF Panel I Control is designed to monitor the presence of 38 CFTR mutations associated with cystic fibrosis, including the 23 mutations recommended for testing by American College of Medical Genetics (ACMG) and American College of Obstetricians and Gynecologists (ACOG). The INTROLTM CF Panel I Control also monitors 3 polymorphisms (1506V, 1507V, F508C) and the 5/7/9T variants.

- 4. Special instrument requirements: Not applicable.
## I. Device Description:

INTROL™ CF Panel I Control consists of synthetic CFTR DNA suspended in a matrix of synthetic DNA targets, carrier DNA of a non-human species, preservatives, dye, and stabilizers. The synthetic DNA contains all 27 CFTR gene exons plus intronic borders, and contains specific mutations and polymorphisms which are divided among 5 bottles (bottles a, b, c, d, and e). The 5 bottles exist in two versions: Version G106ac includes bottles (a), (b), and (c), while Version G106de includes bottles (a), (b), (d), and (e). The specific mutations present in each bottle are listed below in Table 1; all other CFTR sequence is wild type. CFTR mutations that are not listed cannot be detected in the INTROL™ CF Panel I Control.

Control sequences include the mutations, wild type alleles, and polymorphisms recommended for testing by the American College of Medical Genetics (ACMG) and American College of Obstetricians and Gynecologists (ACOG). CFTR mutations that are not listed cannot be detected in the INTROL™ CF Panel I Control.

CFTR DNA is stabilized in the matrix and released when processed through common extraction methods as if it were a whole blood specimen. Following extraction, the released DNA can be used in common amplification based molecular assays techniques. Because INTROL™ CF Panel I Control is designed to mimic the whole blood sample, the resulting copy number of the artificial CFTR segment, after extraction, will be similar to that found in a processed human whole blood sample (v/v).

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Table 1. Composition of INTROLTM CF Panel I Control includes following combinations of CFTR mutations and polymorphisms (plus wild type sequence covering 27 CFTR exons).

| Allele | Genotype | Allele | Genotype | |
|--------------|----------------------------|------------|--------------------------|-------------------|
| | Bottle a (both versions) | | Bottle b (both versions) | |
| 7T* | 7T / 7T | E60X | Homozygous mutant | |
| (I507V)* | I507V / WT | G85E* | Homozygous mutant | |
| (F508C)* | F508C / WT | I148T | Homozygous mutant | |
| S549N/ S549R | Heterozygous | 621+1G>T* | Homozygous mutant | |
| S1251N | Heterozygous | 711+1G>T* | Homozygous mutant | |
| | | 1078delT | Homozygous mutant | |
| | Bottle c (Version ac only) | R334W* | Homozygous mutant | |
| 394delTT | Heterozygous | R347P* | Homozygous mutant | |
| R117H* | Heterozygous | 9T* | 9T / 9T | |
| R347H | Heterozygous | A455E* | Homozygous mutant | |
| 5T* / 7T* | Heterozygous | del F508* | Homozygous mutant | |
| (I506V)* | I506V / WT | V520F | Homozygous mutant | |
| del 1507* | Heterozygous | 1717-1G>A* | Homozygous mutant | |
| R553X* | Heterozygous | G542X* | Homozygous mutant | |
| 2183AA>G | Heterozygous | G551D* | Homozygous mutant | |
| | | R560T* | Homozygous mutant | |
| | Bottle d (Version de only) | 1898+1G>A* | Homozygous mutant | |
| 394delTT | Homozygous mutant | 2143delT | Homozygous mutant | |
| R117H* | Homozygous mutant | 2184delA* | Homozygous mutant | |
| R347H | Homozygous mutant | 2789+5G>A* | Homozygous mutant | |
| 5T* | 5T / 5T | 3120+1G>A* | Homozygous mutant | |
| del 1507* | Homozygous mutant | 3199del6 | Homozygous mutant | |
| R553X* | Homozygous mutant | D1152H | Homozygous mutant | |
| 2183AA>G | Homozygous mutant | R1162X* | Homozygous mutant | |
| | | 3659delC* | Homozygous mutant | |
| | Bottle e (Version de only) | | 3849+10kbC>T* | Homozygous mutant |
| 7T* | 7T / 7T | 3876delA | Homozygous mutant | |
| (I506V)* | I506V / I506V | 3905insT | Homozygous mutant | |
| | | W1282X* | Homozygous mutant | |
| | | N1303K* | Homozygous mutant | |

*ACMG / ACOG Panel

# J. Substantial Equivalence Information:

- 1. Predicate device name(s): Not applicable.
- 2. Predicate 510(k) number(s): Not applicable.
- 3. Comparison with predicate: Not applicable

K. Standard/Guidance Document Referenced (if applicable): None referenced.

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## L. Test Principle:

Not applicable.

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

- 1. Analytical performance:
All INTROL™ CF Panel I Control products are tested by an FDA-cleared CFTR mutation detection method before being released for market distribution. Any mutations not tested by the FDA-cleared method are sequenced bidirectionally before product is released. All mutations must be detected.

- a. Precision/Reproducibility:
The clinical study performed at the external sites evaluated reproducibility of INTROL™ CF Panel I quality control material with respect to within run, between run, between sites, between lots, and between methods. External site evaluations:

The clinical laboratory study performed at the external sites evaluated reproducibility of INTROL™ CF Panel I Control quality control material with respect to within run, between run, between sites, between lots, and between methods.

Evaluation using different extraction methods:

| # extraction<br>methods | #<br>laboratories | # successful laboratory<br>extractions | percent<br>successful |
|-------------------------|-------------------|----------------------------------------|-----------------------|
| 21 | 134 | 129 | 96% * |

* Five laboratories didn't continue after DNA extraction because DNA quantitation method they used indicated that no DNA was extracted/isolated. Considering that the level of synthetic CFTR DNA present in the extracted control may not be detectable with certain quantitation methods, there is a possibility that extractions in these 5 laboratories may have been successful; however this could not be assessed because the assays were not performed.

INTROL™ CF Panel I quality control material has been tested using CFTR assays at 10 external sites. 8 of which were clinical laboratories representing intended user. Samples from 11 different manufacturing lots were tested at minimally 3 external sites in at least 3 separate runs. Results are summarized in the Table 2.

| Method | Site | # of<br>Lots1 | # of<br>Runs | Total<br>Calls | Percent<br>Correct<br>Calls |
|--------------------------------|------|---------------|--------------|----------------|-----------------------------|
| Tag-ItTM | 1 | 10 | 9 | 223 | 100% |
| Tag-ItTM | 2 | 3 | 9 | 138 | 100% |
| Tag-ItTM | 3 | 1 | 1 | 6 | 100% |
| Tag-ItTM | 4 | 1 | 1 | 7 | 100% |
| Tag-ItTM | 5 | 1 | 1 | 4 | 100% |
| eSensor | 6 | 5 | 1 | 30 | 100% |
| eSensor | 7 | 5 | 1 | 38 | 100% |
| Other Amplification<br>methods | 8 | 3 | 1 | 31 | 100% |
| Other Amplification<br>methods | 9 | 1 | 2 | 7 | 100% |
| Other Amplification<br>methods | 10 | 5 | 40 | 649 | 100% |
| 6 methods | | 11 Lots | 66<br>Runs | 1133<br>Calls | 100% |

Table 2. External site evaluations.

Each bottle is processed independently and has its own lot number.

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- b. Linearity/assay reportable range: Not applicable.
- Traceability, Stability, Expected values (controls, calibrators, or methods): C. Bidirectional sequencing of INTROL™ CF Panel I DNA is used to validate the presence of mutant or wild type sequence.

Product acceptance criteria and testing: All INTROL™ CF Panel I Control products are tested by an FDA-cleared CFTR mutation detection method before being released for market distribution. Any mutations not tested by the FDA-cleared method are sequenced bidirectionally before product is released. All mutations must be detected.

Upon receipt and after opening, the material should be stored at 2° - 8°C. INTROL™ CF Panel I Control material is shipped with a "Do not freeze" warning in the device labeling.

Unopened INTROLTM CF Panel I Control material is stable through the expiration date printed on each bottle when stored refrigerated (2° - 8°C). Opened material returned to the refrigerator (2° - 8°C) shortly after use is stable for thirty (30) days from the date of opening.

### Real time stability study:

Results of the real time studies are in the tables below. Stability was tested using 4 different methods, including cleared Tag-It™ Cystic Fibrosis (CF) Kit from Tm Bioscience, and bidirectional sequencing. Testing was performed at 1, 2, 3, 4, and 12 months. In addition, real time testing was performed with 5 different lots of quality control material. At 12 months, all mutations and variants were tested using either the FDA cleared assay or bidirectional sequencing. Data from this ongoing study currently supports a shelf-life of 12 months.

| Stability Testing | Number of<br>Lots | Percent<br>Correct Calls |
|---------------------|-------------------|--------------------------|
| Date of Manufacture | 5 | 100% |
| 1 month | 5 | 100% |
| 2 months | 3 | 100% |
| 3 months | 1 | 100% |
| 4 months | 2 | 100% |
| 12 months | 3 | 100% |

#### Stress testing:

Elevated Temperature Studies:

The study goal was to demonstrate product stability during prolonged shipping without refrigeration. A bottle of INTROLTM CF Panel I was removed from storage and mailed across the US and back without wet ice. Testing of the stressed product in the CFTR assay showed no loss of signal.

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Two samples of INTROL™ CF Panel were compared after one was incubated at 60°C for 21-24 days, and the other incubated at 4°C, extracted and tested in duplicate in CFTR assay. There was no loss of signal after incubation at 60°C.

### Freeze/Thaw Studies:

A bottle of INTROL™ CF Panel I was placed at -20℃ for 48 hours, thawed and tested in CFTR assay. No loss of signal was detectable after one cycle of freeze/ thaw.

### Open Vial Stability:

Two open vial studies were performed. In the first study, a bottle of INTROLTM CF Panel I was mixed, opened briefly, then stored at 2-8℃ for 49 days. In the second study, a bottle of INTROL™ CF Panel I was mixed and opened with pipette simulation six times over 35 days. Both studies demonstrated no loss of signal when used in CFTR assay at the end of the test period.

#### Expected Results:

Expected results with the INTROLTM CF Panel I Control using two FDAcleared CFTR assays are presented in Table 3.

| Method | Correctly Identified Alleles | No Call or other | Not Tested |
|---------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Tag-It | All wt alleles<br>7T, I507V, F508C, S549R, G85E, I148T, 621+1G>T,<br>1078delT, R334W, R347P, 9T, A455E, delF508,<br>V520F, 1717+1G>A, G542X, G551D, R560T,<br>1898+1G>A, 2184delA, 3120+1G>A, R1162X,<br>3659delC, 3849+10kbC>T, 3876delA, 3905insT,<br>W1282X, N1303K, 394delTT, R117H, R347H, 5T/7T,<br>I506, dell507, R553X, 2183AA>G | S549N MUT1<br>711+1G>T No Call2<br>2789+5G>A No Call3 | S1251N, E60X, 2143delT<br>3199del6, D1152H, |
| eSensor | All wt alleles<br>7T, I148T, 621+1G>T, 1078delT, R334W, R347P, 9T,<br>A455E, delF508, 1717+1G>A, G542X, G551D, R560T,<br>1898+1G>A, 2184delA, 2789+5G>A, 3120+1G>A,<br>R1162X, 3659delC, 3849+10kbC>T, W1282X,<br>N1303K, R117H, 5T/7T, dell507, R553X, | G85E No Amp3<br>711+1G>T No Amp4 | I507V, F508C, S549N,<br>S549R, S1251N, E60X,<br>1078delT V520F,<br>2143delT, 3199del6,<br>D1152H, 3876delA,<br>3905insT, 394delTT,<br>R347H, I506V,<br>2183AA>G, |

Table 3. Results with the INTROL™ CF Panel I using two FDA cleared CFTR assays.

' Detected as homozyqous mutant.

Insufficient control sequence for one of the 711+1G>T amplicon primers.

3 Insufficient control sequence for one of the 2789+5G>A amplicon primers.

4 Mutation E60X interferes with a G85E amplicon primer.

- d. Detection limit: Not applicable
- e. Analytical specificity: Not applicable.
- f. Assay cut-off: Not applicable.

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- 2. Comparison studies:
- Method comparison with predicate device: a. Not applicable.
- b. Matrix comparison: Not applicable.
- 3. Clinical studies:
- a. Clinical Sensitivity: Not applicable.
- Clinical specificity: b. 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 petition for Evaluation of Automatic Class III Designation for this device is accepted. 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.