HYBRID CAPTURE II CT-ID TEST

K990023 · Digene Corp. · LSK · Oct 25, 1999 · Microbiology

Device Facts

• Record ID: K990023
• Device Name: HYBRID CAPTURE II CT-ID TEST
• Applicant: Digene Corp.
• Product Code: LSK · Microbiology
• Decision Date: Oct 25, 1999
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3120
• Device Class: Class 1

Indications for Use

The Digene HCII CT-ID Test is an in-vitro nucleic acid hybridization assay with signal amplification using microplate chemiluminescence for the qualitative detection of C trachomatis DNA in cervical specimens collected using the Digene Cervical Sampler™ (Cervical Brush and Speciment Transport Medium) and in cervical specimens collected using the Digene Swab Specimen Collection Kit (Dacron Swab and Speciment Transport Medium). The Digene HCII CT-ID Test is indicated for use with symptomatic or asymptomatic women as evidence of infection with C. trachomatis. The HCII CT-ID Test may be used alone or as a supplemental test to the Digene HCII CT/GC Test to detect C. trachomatis DNA.

Device Story

Nucleic acid, signal-enhanced, hybridization microplate assay; detects C. trachomatis DNA in cervical specimens. Process: specimen denaturation (sodium hydroxide, 65°C) releases DNA; hybridization with RNA probe cocktail; capture of RNA:DNA hybrids on antibody-coated microplate wells; reaction with alkaline phosphatase-conjugated antibody and chemiluminescent substrate. Luminometer measures photon emission as Relative Light Units (RLUs). Used in clinical laboratories; operated by technicians. Output: qualitative RLU measurement interpreted against positive cutoff value. Assists clinicians in diagnosing C. trachomatis infection; enables identification of positive specimens when used as a supplement to the HCII CT/GC test.

Clinical Evidence

Multicenter study (N=940) compared HCII CT-ID to cell culture/DFA. Overall sensitivity 97.25% (95% CI: 92.2-99.4) and specificity 98.07% (95% CI: 96.9-98.9). Additional site-specific data provided for symptomatic and asymptomatic populations. Bench testing confirmed probe specificity and DNA stabilization in transport medium.

Technological Characteristics

Nucleic acid hybridization assay; chemiluminescence detection. Components: RNA probe cocktail, alkaline phosphatase-conjugated antibodies, microplate wells coated with anti-RNA:DNA hybrid antibodies. Connectivity: interfaces with DML 2000 luminometer. Software-based RLU/cutoff ratio calculation.

Indications for Use

Indicated for symptomatic or asymptomatic women to aid in the diagnosis of Chlamydia trachomatis infection. Specimens are cervical samples collected via brush or swab.

Regulatory Classification

Identification

Chlamydia serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to chlamydia in serum. Additionally, some of these reagents consist of chlamydia antisera conjugated with a fluorescent dye used to identify chlamydia directly from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Chlamydia and provides epidemiological information on these diseases. Chlamydia are the causative agents of psittacosis (a form of pneumonia), lymphogranuloma venereum (a venereal disease), and trachoma (a chronic disease of the eye and eyelid).

Predicate Devices

• Gen-Probe Pace 2 System for Chlamydia Trachomatis (K920378)

Reference Devices

• Digene HCII CT/GC Test
• DML 2000 Microplate Luminometer

Submission Summary (Full Text)

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# OCT 2 5 1999

### 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: N/A - This is a new 510(k)

### Submitter's Name, Address, Telephone Number, Contact Person and Date Prepared

Submitter Digene Corporation 9000 Virginia Manor Road Beltsville, MD 20705

[phone] Phone: Facsimile: [phone]

Contact Person Bryan Schneider, Regulatory Specialist Digene Corporation Phone: [phone] Facsimile: [phone]

Date Prepared: 1/4/99

### Name of Device and Name/Address of Sponsor

Name of Device Digene Hybrid Capture® II CT-ID Test

Sponsor Digene Corporation 9000 Virginia Manor Road Beltsville, MD 20705 Tel: [phone] Fax: [phone]

Common or Usual Name

HCII CT-ID Test

A28-1

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### Classification Name

### DNA Reagents, Chlamydia

### Predicate Device(s)

The Gen-Probe Pace 2 System for Chlamydia Trachomatis cleared under K920378 on 4/29/92.

### Device Description

The Digene HCII CT-ID Test is a nucleic acid, signal enhanced, hybridization, microplate assay using chemiluminescence for the qualitative detection of C. trachomatis (CT) DNA in cervical specimens collected using the Digene Cervical Sampler™ and in cervical specimens collected with a Dacron swab and placed in Digene Specimen Transport Medium. The Digene HCII CT-ID Test is indicated for use as an aid in diagnosing infection with C. trachomatis in symptomatic or asymptomatic women. The HCII CT-ID Test may be used as a stand-alone test or may be used as a supplemental test to the Digene HCII CT/GC Test for identification of C. trachomatis in specimens that are positive by the HCII CT/GC Test.

Specimens potentially containing CT DNA are denatured and then hybridized with a specific RNA probe cocktail. This cocktail contains a probe mixture chosen to minimize or eliminate cross-reactivity with DNA sequences from human cells, other bacterial species, Chlamydia species other than trachomatis, or sequences from other operior misms common in urogenital specimens. The CT probe cocktail supplied with the Digene CT-ID Assay is complementary to approximately 39,300 base pairs or 4% of the C. trachomatis genome (1 x 10 base pairs) and 100% of the cryptic plasmid.

### Steps and Reagents to Stabilize the Specimen:

Specimens are collected using the Digene Cervical Sampler or a Dacron Swab and placed in Digene Specimen Transport Medium (STM). The specimens may be held for up to two weeks at room temperature and shipped without refrigeration to the testing laboratory in an insolated container using an overnight or 2-day delivery vendor. At the testing laboratory specimens should be stored at 2-8℃ if the assay is to be performed within one week. If the assay will be performed later than one week, the specimens should be stored at -20°C. A preservative has been added to the STM to retard bacterial growth and retain the integrity of DNA in the specimen. The STM is not intended to preserve the viability of organisms or other cells in the specimen.

### Steps to Process the Specimen, Release the DNA, and Denature the Released DNA.

Denaturation reagent is added to the specimens with the collection device remaining in the collection tube. This allows for denaturation of any DNA clinging to the collection device. The volume of denaturation reagent added to the specimen is equivalent to one-half the volume of the specimen. The denaturation reagent is dilute sodium hydroxide. Specimens are then incubated for 45 minutes at 65°C. This step releases the DNA from the organisms contained in the specimen and denatures that DNA so it becomes single-stranded. Following this step, the single-stranded DNA is ready to be hybridized to the RNA probe.

¹ Kingsbury DT. Estimate of the genome size of various microorganisms. J Bacteriol 1969 Jun:98(3):1400-1.

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The RNA:DNA hybrids resulting from hybridization are immobilized (captured) on the surface of a microplate-well, which has been coated with antibodies specific for RNA:DNA hybrids. The antibodies on the well surface capture the RNA:DNA hybrids. The immobilized hybrids are then reacted with alkaline phosphatase-conjugated antibody and a chemiluminescent substrate. As the substrate is cleaved by the bound alkaline phosphatase, photons are emitted and measured as Relative Light Units (RLUs) using a standard, FDA-cleared luminometer such as the DML 2000™ - Increased photon emission, resulting in an enhanced signal, is achieved by conjugating multiple alkaline phosphatase molecules to each antibody molecule. Multiple antibodies bind to each RNA:DNA hybrid, further enhancing the signal.

The HCII CT-ID Test provides an RLU measurement that is qualitatively interpreted. The Positive Cutoff Value is equal to the mean of three Positive Control values. Each specimen RLU measurement is converted to a ratio of the Positive Cutoff Value. This conversion calculation is performed automatically by the Digene DML™ 2000 Microplate Luminometer software. Alternatively, the conversion may be calculated manually, Specimens with RLU/Cutoff values of < 0.8 are considered negative for CT DNA. Specimens with RLU/Cutoff values >5.0 are considered positive for CT DNA. Specimens with RLU/Cutoff values between 0.8 ≥ 5.0 are considered to be equivocal and are repeat tested in duplicate. With the repeat tests, a RLU/Cutoff Value of 1.0 is applied. If two of the three replicates fall above 1.0, the presence of C. trachomatis DNA is indicated. If at least two of the three replicates fall below 1.0, the presence of C. trachomatis DNA is not indicated.

### Intended Use

The Digene HCII CT-ID Test, is a nucleic acid, signal enhanced, hybridization, microplate assay for the qualitative detection of C. trachomatis DNA in cervical specimens collected using the Digene Cervical Sampler™, or collected using a Dacron® swab and placed in Digene Specimen Transport Medium. The Digene HCII CT-ID Test is indicated for use as an aid in diagnosing infection with C. trachomatis in symptomatic or asymptomatic females. The HCII CT-ID Test may be used as a stand-alone test or may be used as a supplemental test for identification of C. trachomatis in specimens found positive by the Digene HCII CT/GC Test.

### Technological Characteristics and Substantial Equivalence

The Digene HCll CT-ID Test is substantially equivalent to the Gen-Probe Pace 2 System for C. trachomatis in intended use and in technological characteristics. Both tests are nucleic acid hybridization assays intended for the detection of chlamydia trachomatis from endocervical specimens.

A multicenter study has demonstrated that the Digene HCII CT-ID Test performs as well or better than the gold standard, cell culture, in detecting infection with C. trachomatis in the intended population. The results from this multicenter study are summarized below:

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# HCll CT-ID Test versus CT Culture/DF/

| CT-ID | | | Culture | | | DFA:<br>NA | N | Site | 2x2 Table<br>Reference | Prevalence<br>(%) | Sensitivity<br>(95% CI) | Specificity<br>(95% CI) | Ct-ID+/Cul-<br>/DFA-<br>Tested Positive<br>by PCR | NPV<br>(95% CI) | PPV<br>(95% CI) |
|-------|-----|------------|---------|-----|------------------|------------|-----|------|------------------------|-------------------|-------------------------|-------------------------|---------------------------------------------------|----------------------|----------------------|
| POS | NEG | POS<br>NEG | POS | NEG | POS<br>NEG<br>NA | | | | | | | | | | |
| | | | | | | | 351 | UAB | 18 | 13.96 | 95.92<br>(86.0-99.5) | 97.68<br>(95.3-99.1) | 5/7 | 99.33<br>(97.6-99.9) | 87.04<br>(75.1-94.6) |
| | | | | | | | 192 | JHU | 19 | 8.33 | 100.00<br>(79.4-100) | 96.59<br>(92.7-98.7) | 6/6 | 100.00<br>(97.9-100) | 72.73<br>(49.8-89.3) |
| | | | | | | | 220 | SUNY | 20 | 15.91 | 97.14<br>(85.1-99.3) | 98.38<br>(95.3-99.7) | 1/2 | 99.45<br>(97.0-100) | 91.89<br>(78.1-98.3) |
| | | | | | | | 177 | UCSF | 21 | 5.08 | 100.00<br>(66.4-100) | 100.00<br>(97.8-100) | NA | 100.00<br>(97.8-100) | 100.00<br>(66.4-100) |
| | | | | | | | 940 | All | 22 | 11.60 | 97.25<br>(92.2-99.4) | 98.07<br>(96.9-98.9) | 12/15 | 99.63<br>(98.9-99.9) | 86.89<br>(79.6-92.3) |

DO NOT DELAY - FILE IMMEDIATELY
NICS WAS TERMINATED
7/1/2021
ONE FORM PER TRANSACTION

- One CT-ID negative, culture negative specimen was unnecessarily tested by DFA and gave a positive result. This result was included in the pen
calculations as a HCILCT-ID false negative.
ce

t hwc case, DFA was required with in the love a positive sest. The patim andred in its perfornance
all chil negative, ciller negative by clime and negative by DFA. The t

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| x2 Table<br>Reference | Site | CT-ID:<br>Culture: | DFA:<br>N | POS<br>POS<br>NA | POS<br>NEG<br>POS | POS<br>NEG<br>NEG | NEG<br>POS<br>NA | NEG<br>NEG<br>POS | NEG<br>NEG<br>NEG | Prevalence<br>(%) | Sensitivity<br>(95% CI) | Specificity<br>(95% CI) | Ct-ID+/Cul-<br>/DFA-<br>Tested Positive<br>by PCR | NPV<br>(95% CI) | PPV<br>(95% CI) |
|------------------------|------|--------------------|------------|------------------|-------------------|-------------------|------------------|-------------------|-------------------|-------------------------|-------------------------|---------------------------------------------------|---------------------------------------------------|----------------------|----------------------|
| 23 | UAB | 101 | 8 | 0 | 2 | 0 | NA | 91 | | 7.92 | 100.00<br>(63.1-100) | 97.85<br>(92.5-99.7) | 0/2 | 100.00<br>(96.0-100) | 80.00<br>(44.4-97.5) |
| 24 | JHU | 12 | 1 | 0 | 1 | 0 | NA | 10 | | 8.33 | 100.00<br>(2.50-100) | 90.91<br>(58.7-99.8) | 1/1 | 100.00<br>(69.2-100) | 50.00<br>(1.3-98.7) |
| 25 | SUNY | 81 | 3 | 0 | 0 | 0 | NA | 78 | | 3.70 | 100.00<br>(29.2-100) | 100.00<br>(95.4-100) | NA | 100.00<br>(95.4-100) | 100.00<br>(29.2-100) |
| 26 | UCSF | 236 | 9 | 1 | 4 | 0 | NA | 222 | | 4.24 | 100.00<br>(69.2-100) | 98.23<br>(95.5-99.5) | 3*/4 | 100.00<br>(98.4-100) | 71.43<br>(41.9-91.6) |
| 27 | SJH | 1 | 0 | 0 | 0 | 0 | NA | 1 | | 0 | N/A | 100.00<br>(2.5-100) | | 100.00<br>(2.5-100) | N/A |
| 28 | All | 431 | 21 | 1 | 7 | 0 | NA | 402 | | 5.10 | 100.00<br>(84.6-100) | 98.29<br>(96.5-99.3) | 4*/7 | 100<br>(99.1-100) | 75.86<br>(56.5-89.7) |
| | | | CT-ID: | | | | | | | | | | | | |
| | | | Culture: | | | | | | | | | | | | |
| | | | DFA: | | | | | | | | | | | | |
| 2x2 Table<br>Reference | Site | N | POS<br>POS | POS<br>NEG | POS<br>NA | NEG<br>POS | NEG<br>NEG | NEG<br>NA | Prevalence<br>(%) | Sensitivity<br>(95% CI) | Specificity<br>(95% CI) | Ct-ID+/Cul-<br>/DFA-<br>Tested Positive by<br>PCR | NPV<br>(95% CI) | PPV<br>(95% CI) | |
| 35 | UAB | 7 | 2 | 0 | 0 | 1 | 4 | NA | 42.86 | 66.67<br>(94.3-99.2) | 100<br>(39.8-100) | N/A | 80.00<br>(28.4-99.5) | 100.00<br>(15.8-100) | |
| 36 | JHU | 94 | 10 | 1 | 3 | 1 | 79 | NA | 12.77 | 91.67<br>(61.5-99.8) | 96.34<br>(89.6-99.2) | 2/3 | 98.75<br>(93.2-99.9) | 78.57<br>(49.2-95.3) | |
| 37 | SUNY | 8 | 1 | 0 | 0 | 2 | 5 | NA | 37.50 | 33.33<br>(0.84-90.6) | 100.00<br>(47.8-100) | N/A | 71.43<br>(29.0-96.3) | 100.00<br>(2.5-100) | |
| 38 | SJH | 152 | 7 | 0 | 2 | 0 | 143 | NA | 4.61 | 100.00<br>(59.0-100) | 98.62<br>(95.1-99.8) | 0/1 | 100.00<br>(97.5-100) | 77.78<br>(40.0-97.2) | |
| 39 | All | 261 | 20 | 1 | 5 | 4 | 231 | NA | 9.58 | 84.00<br>(63.9-95.5) | 97.88<br>(95.1-99.3) | 2/4 | 98.30<br>(95.7-99.5) | 80.77<br>(60.7-93.5) | |

- Bone case, PCR was not done.

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# HCli CT-ID Test versus CT Culture/DFA

In one case DFA was required but not done.

In one case, PCR was not done.

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| HCII CT-ID Test versus CT Culture/DFA |
|---------------------------------------|
| Asymptomatic Patients |
| Dacron Swab Only |

| | | CT-ID: | POS | POS | POS | NEG | NEG | | | | | | |
|-----------------------|------|-----------|-----|-----|-----|-----|-----|-------------------|-------------------------|-------------------------|--------------------------------------------|----------------------|----------------------|
| | | Culture: | POS | NEG | NEG | POS | NEG | | | | | | |
| x2 Table<br>Reference | Site | DFA:<br>N | POS | NEG | POS | NEG | NEG | Prevalence<br>(%) | Sensitivity<br>(95% CI) | Specificity<br>(95% CI) | CI-ID+CuHDFA-<br>Tested Positive by<br>PCR | NPV<br>(95% CI) | PPV<br>(95% CI) |
| 40 | UAB | 1 | 1 | 0 | 0 | 0 | 0 | 100 | 100<br>(2.5-100) | NA | N/A | NA | 100.00<br>(2.5-100) |
| 41 | JHU | 10 | 0 | 0 | 0 | 0 | 10 | 0.0 | NA | 100<br>(69.2-100) | N/A | 100.00<br>(69.2-100) | NA |
| 42 | SUNY | 2 | 0 | 0 | 1 | 0 | 1 | 0.0 | NA | 50.00<br>(1.3-98.7) | N/A | 100.00<br>(2.5-100) | 0.00<br>(0-97.5) |
| 43 | UCSF | 1 | 0 | 0 | 0 | 0 | 1 | 0.0 | NA | 100<br>(2.5-100) | N/A | 100.00<br>(2.5-100) | NA |
| 44 | SJH | 176 | 2 | 0 | 0 | 0 | 174 | 1.14 | 100.00<br>(15.8-100) | 100.00<br>(97.9-100) | N/A | 100.00<br>(97.9-100) | 100.00<br>(15.8-100) |
| 45 | All | 190 | 3 | 0 | 1 | 0 | 186 | 1.58 | 100.00<br>(29.2-100) | 99.47<br>(97.1-100) | N/A | 100.00<br>(98.0-100) | 75.00<br>(19.4-99.4) |

CT/GC+/Cul- = specimens positive by Hybrid Cres and negative by CH. The data represented in this table are
unresolved. The PCR data is provided for informational proses onl

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Image /page/7/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular in shape and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter of the circle. Inside the circle is an abstract symbol that resembles an eagle or other bird-like figure.

## OCT 2 5 1999

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Mr. Mark A. Del Vecchio Associate Director, Regulatory and Clinical Affairs Digene Corpration 9000 Virginia Manor Road Rockville, Maryland 20705

Re: K990023 Trade Name: Digene HCII CT-ID Test Regulatory Class: I Product Code: LSK Dated: August 16, 1999 Received: August 17, 1999

Dear Mr. Del Vecchio:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (OS) for Medical Devices: General regulation (21 CFR Part 820) and that. through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device Onder the Omical Baoordexity categorization. To determine if it ides, you should contact a the Centers for Disease Control and Prevention (CDC) at [phone].

This letter will allow you to begin marketing your device as described in your 510(k) I mis reter will anow Jour of Substantial equivalence of your device 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 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and Compliance at (301) 5947 price, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Autman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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Page_I_of_I

510(k) Number (if known):_K 99 co 2 3 Device Name: Digene HC II CT - ID

Device Name: *Digene's HC II CT -*

Indications For Use:

The Digene HCII CT-ID Test is an in-vitro nucleic acid hybridization assay with signal amplification using microplate chemiluminescence for the qualitative detection of C trachomatis DNA in cervical specimens collected using the Digene Cervical Sampler™ (Cervical Brush and Speciment Transport Medium) and in cervical Sampler™
using the Digene Swah Speciment Collection (Aind) specimens collected using the Digene Swab Specimen Collection Kit (Dacron) Swab Specimens Collected
Transport Medium) The Digene Transport Medium). The Digene HCII Onlietion Swab and Speciment
or asymptomatic women as evidence of infrastian with CT-ID Test is indicated for use with symptomatic or used to be and the Digene Holl CT4D Test is indicated for use

The HCII CT-ID Test may be used alone or as a supplemental test to the Digene HCII CT/GC Test to detect C. trachomatis Dras a Supplemental test to the Digene HCll
CT/GC Test. CT/GC Test.

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Wooley Dubois

(Division Sign Off)
Division of Clinical Laboratory Devices
510(k) Number K990023

Prescription Use
(Per 21 CFR 801.109)

OR

Over-The-Counter Use__________________________________________________________________________________________________________________________________________________________

(Optional Format 1-2-96)