OC-Light S FIT

K143325 · Eiken Chemical Co., Ltd. · KHE · Aug 20, 2015 · Hematology

Device Facts

• Record ID: K143325
• Device Name: OC-Light S FIT
• Applicant: Eiken Chemical Co., Ltd.
• Product Code: KHE · Hematology
• Decision Date: Aug 20, 2015
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.6550
• Device Class: Class 2

Intended Use

OC-Light S FIT (Fecal Immunochemical Test, also known as iFOBT, immunochemical fecal occult blood test) is a qualitative test intended for the immunochemical detection of fecal occult blood (FOB) by professional laboratories and physician office laboratories. Measurement of FOB is useful as an aid to detect blood in stool when gastrointestinal (GI) bleeding may be suspected. OC-Light S FIT is recommended for use in routine physical examinations.

Device Story

OC-Light S FIT is an in vitro diagnostic lateral flow chromatographic immunoassay for qualitative detection of human hemoglobin (hHb) in stool. The device consists of a test strip and a sampling bottle containing 2.0 mL extraction buffer. The test strip features a nitrocellulose membrane with immobilized mouse monoclonal anti-hHb antibodies at the test region and rabbit anti-mouse antibodies at the control region; a conjugate pad contains colloidal gold-conjugated mouse monoclonal anti-hHb antibodies. When the sample end is dipped in fecal extract, hHb binds to the gold conjugate; the complex migrates to the test region, forming a visible reddish/pink line if hHb is present. A procedural control line confirms proper wicking and technique. The test is performed by healthcare professionals in laboratories or physician offices. Results are visually interpreted by the operator. A positive result indicates the presence of hHb above the 10 µg/g stool cut-off, aiding in the suspicion of GI bleeding. The device provides a non-invasive screening tool for GI disorders.

Clinical Evidence

Bench testing only. Performance evaluated via repeatability and reproducibility studies across three sites (n=1764 total results), demonstrating high agreement. Analytical specificity confirmed against animal hemoglobins, meat/vegetable extracts, iron, sodium L-ascorbate, and toilet cleaners. No susceptibility to prozone/hook effect up to 2000 ng/mL. Method comparison against predicate (K041297) showed high overall percent agreement (99.7% in POLs, 100% in PMLs).

Technological Characteristics

Lateral flow chromatographic immunoassay. Components: nitrocellulose membrane, sample pad, conjugate pad (colloidal gold-conjugated mouse monoclonal anti-hHb), absorption pad, plastic backing. Sampling bottle contains 2.0 mL extraction buffer. Qualitative visual readout. No electronic components, software, or connectivity. Sterilization not specified. Cut-off: 10 μg hemoglobin/g stool (50 ng/mL).

Indications for Use

Indicated for use in routine physical examinations for the detection of fecal occult blood in stool specimens to aid in the identification of gastrointestinal bleeding in patients.

Regulatory Classification

Identification

An occult blood test is a device used to detect occult blood in urine or feces. (Occult blood is blood present in such small quantities that it can be detected only by chemical tests of suspected material, or by microscopic or spectroscopic examination.)

Special Controls

*Classification.* Class II (special controls). A control intended for use with an occult blood test is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 864.9.

Predicate Devices

• Polymedco OC-Light FOB Test (K041297)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, stacked on top of each other.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 20, 2015

EIKEN CHEMICAL CO., LTD. Takashi Enomoto, Regulatory Affairs 4-19-9, Taito, Taito-ku Tokyo 110-8408, Japan

Re: K143325

Trade/Device Name: OC-Light S FIT Regulation Number: 21 CFR 864.6550 Regulation Name: Occult blood test Regulatory Class: Class II Product Code: KHE Dated: July 15, 2015 Received: July 16, 2015

Dear Mr. Enomoto:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. 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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set

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forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

# Leonthena R. Carrington -S

Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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| DEPARTMENT OF HEALTH AND HUMAN SERVICES<br>Food and Drug Administration<br><b>Indications for Use</b> | Form Approved: OMB No. 0910-0120<br>Expiration Date: January 31, 2017<br>See PRA Statement below. |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 510(k) Number (if known) | K143325 |
| Device Name | OC-Light S FIT |
| Indications for Use (Describe) | OC-Light S FIT (Fecal Immunochemical Test, also known as iFOBT, immunochemical fecal occult blood test) is a<br>qualitative test intended for the immunochemical detection of fecal occult blood (FOB) by professional laboratories and<br>physician office laboratories. Measurement of FOB is useful as an aid to detect blood in stool when gastrointestinal (GI)<br>bleeding may be suspected.<br>OC-Light S FIT is recommended for use in routine physical examinations. |
| Type of Use (Select one or both, as applicable) | <label><input checked="checked" type="checkbox"/> Prescription Use (Part 21 CFR 801 Subpart D)</label> <label><input type="checkbox"/> Over-The-Counter Use (21 CFR 801 Subpart C)</label> |
| PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED. | |
| FOR FDA USE ONLY | |
| Concurrence of Center for Devices and Radiological Health (CDRH) (Signature) | |
| This section applies only to requirements of the Paperwork Reduction Act of 1995.<br>*DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* | |
| The burden time for this collection of information is estimated to average 79 hours per response, including the<br>time to review instructions, search existing data sources, gather and maintain the data needed and complete<br>and review the collection of information. Send comments regarding this burden estimate or any other aspect<br>of this information collection, including suggestions for reducing this burden, to: | |
| Department of Health and Human Services<br>Food and Drug Administration<br>Office of Chief Information Officer<br>Paperwork Reduction Act (PRA) Staff<br>[e-mail] | |
| "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of<br>information unless it displays a currently valid OMB number." | |
| FORM FDA 3881 (1/14) | Page 1 of 1 PSC Publishing Services [phone] EF |

and the comments of the state of the states of the states of

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## "510(k) Summary"

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

## 510(k) Submitter Information:

| Name: | EIKEN CHEMICAL CO., LTD |
|----------------------|---------------------------------------------------------------------------------|
| Address: | 4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan |
| Phone: | 81-3-58463479 |
| Fax: | 81-3-58463474 |
| Contact Name & Info: | Takashi Enomoto<br>Regulatory Affairs Department<br>Takashi [e-mail] |

Date Prepared: August 20, 2015

## Device Information:

| Trade Name: | OC-Light S FIT |
|----------------------|--------------------------------------------------------|
| Common Name: | Fecal Immunochemical Test |
| Classification Name: | Occult Blood Test (21 CFR 864.6550, Product Code: KHE) |
| Device Class: | Class II |
| 510(k) Number: | K143325 |
| Manufacturer: | Eiken Chemical Co., Ltd. |

## Predicate Device Information:

| Trade Name: | Polymedco OC Light FOB Test |
|-----------------------|------------------------------------------------------------------------|
| 510(k) Number: | K041297 |
| 510(k) Applicant: | Polymedco Inc.<br>(currently Polymedco Cancer Diagnostic Products LLC) |
| Manufacturer: | Eiken Chemical Co., Ltd. |
| Repackager/Relabeler: | Polymedco Cancer Diagnostic Products LLC |

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Image /page/4/Picture/0 description: The image shows the logo of Eiken Chemical Co., LTD. The logo includes a circular graphic with three dots connected by lines, followed by the company name and address. The address is 4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan, and the website is http://www.eiken.co.jp.

## Device Description:

OC-Light S FIT is an in vitro diagnostic device, a qualitative test designed for the immunochemical detection of human hemoglobin (hHb) in stool specimens.

OC-Light S FIT consists of a test strip, and a sampling bottle containing buffer solution. OC-Light S FIT test strip consists of a nitrocellulose membrane with immobilized mouse monoclonal antibodies specific to human hemoglobin at the test region and immobilized rabbit anti-mouse antibodies at the control region, a sample pad, a conjugate pad which contains human hemoglobin specific mouse monoclonal antibodies conjugated with colloidal gold, an absorption pad, and a plastic backing. OC-Light S FIT sampling bottle is a plastic bottle for collecting fecal sample containing 2.0 mL of extraction buffer.

When the sample end of the test strip is dipped in the fecal extract, the liquid fecal extract wicks through a series of absorbent materials and contacts colloidal gold conjugated with monoclonal antibodies specific to hHb. If hHb is present in the sample, it reacts with the antibodies on the colloidal gold. When the gold conjugate with hHb reaches the test region of the membrane, it binds with the immobilized antibodies also specific to hHb to form a visible reddish/pink line. The procedural control region of the membrane contains immobilized anti-mouse antibodies that capture the conjugate independent of the presence of the hHb, thereby always producing a distinct reddish/pink line. The reddish/pink line in the procedural control region demonstrates the validity of the test, and assures the operator that the device is working properly.

#### Clinical cut-off

The clinical cut-off of OC-Light S FIT is 10 µg hemoglobin /g stool. (50 ng/mL buffer)

#### Controls

Internal Control: Procedural controls are included in the test device. A reddish/pink line appearing in the control region is serves as the internal procedural control. It confirms sufficient specimen volume, adequate membrane wicking and correct procedural technique.

External Control: FOBT-CHEK positive and negative controls, cleared under 510(k) 041297, are available in a separate control kit. The controls are run in the FIT in the same manner as extracted fecal samples. It is recommended that positive and negative controls be performed to verify proper test performance.

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Image /page/5/Picture/0 description: The image shows the logo for EIKEN CHEMICAL CO., LTD. The logo consists of a circular graphic to the left of the company name and address. The address is 4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan. The website address is http://www.eiken.co.jp.

## Intended Use:

OC-Light S FIT (Fecal Immunochemical Test, also known as iFOBT, immunochemical fecal occult blood test) is an qualitative test intended for the immunochemical detection of fecal occult blood (FOB) in feces by professional laboratories and physician office laboratories. Measurement of FOB is useful as an aid to detect blood in stool when gastrointestinal (GI) bleeding may be suspected.

OC-Light S FIT is recommended for use in routine physical examinations.

## Device Characteristics in Comparison with the Predicate Device:

The table below provides a summary of the device characteristics for OC-Light S FIT and the predicate device.

| Device | OC-Light S FIT | Polymedco OC Light FOB Test<br>(Predicate) |
|------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Similarities | | |
| Intended Use | A qualitative test intended for the<br>immunochemical detection of fecal<br>occult blood in feces by<br>professional laboratories and<br>physician office laboratories.<br>Measurement of FOB is useful as<br>an aid to detect blood in stool<br>when gastrointestinal (GI) bleeding<br>may be suspected. | An immunological test intended<br>for the detection of fecal occult<br>blood in feces by professional<br>laboratories and physician office<br>labs The test is useful for the<br>determination of gastrointestinal<br>(GI) bleeding, found in a number<br>of gastrointestinal (GI) disorders,<br>e.g., diverticulitis, colitis, polyps,<br>and colorectal cancer. |
| Indications for<br>Use | Recommended for use in routine<br>physical examinations. | Recommended for use in 1) routine<br>physical examinations, 2)<br>monitoring for bleeding in<br>patients, and 3) screening for<br>colorectal cancer or<br>gastrointestinal bleeding. |
| Sample Type | Feces in an extraction buffer | Feces in an extraction buffer |
| Test Principle | Qualitative test system intended<br>for immunochemical detection of<br>fecal occult blood in feces. | Immunological test system<br>intended for qualitative detection<br>of fecal occult blood in feces. |
| Detection<br>Method | Lateral flow chromatographic<br>immunoassay | Lateral flow chromatographic<br>immunoassay |
| Detection Level | 10µg hemoglobin/g stool<br>(50ng/ml buffer) | 50ng/mL hHb in fecal extraction<br>buffer |

#### Comparison of technological characteristics

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CAL CO.. LTD

-ku. Tokvo 110-8408. Japan

| Differences | | |
|------------------------------|------------------------------------------------------------------------|----------------------------------------------------------------|
| Detection Antibody Conjugate | Monoclonal anti-hHb conjugated to colloidal gold | Monoclonal and polyclonal anti-hHb conjugated to blue latex |
| Result Format | Visible reddish/pink line in procedural control region and test region | Visible blue line in procedural control region and test region |

## Summary of Performance Data:

#### Cut-off

Cut-off study was conducted in-house with one operator. Hb-free stool specimen was spiked with known levels of hHb to obtain fecal samples with seven different Hb concentrations: 0, 5, 8, 10, 12, 15, and 400 µg/ g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, clinical cutoff), 60 (C95+20%), 75 and 2000 ng/mL. Each sample was collected with twenty one OC-Light S FIT sampling bottles and twenty one Polymedco OC Light FOB Test (Predicate) sampling bottles to perform side-by-side tests for comparing the test results of the device with that of the predicate.

The test result of 50ng/mL using OC-Light S FIT was 100.0% positive. The overall agreement between the total results of OC-Light S FIT and the expected results was 99.3% (95% CI 96.2 ~ 100.0%), and the overall agreement between the total results of Polymedco OC Light FOB Test and the expected results was 97.3% (95% CI 93.1 ~ 99.3%)

The cut-off value of OC-Light S FIT was established as 10 µg hemoglobin/g stool (50 ng/mL buffer). No significant difference was observed between the results of OC-Light S FIT and the predicate Polymedco OC Light FOB Test.

| Cut off study | Agreement | | Expected Results | | | Overall<br>Percent<br>Agreement<br>(95% CI) | Positive<br>Percent<br>Agreement<br>(95% CI) | Negative<br>Percent<br>Agreement<br>(95% CI) |
|-----------------------------------|------------------|----|---------------------|---------------------|-----------------------------|---------------------------------------------|----------------------------------------------|----------------------------------------------|
| | | | Positive<br>Results | Negative<br>Results | Total<br>Results | | | |
| Polymedco<br>OC Light<br>FOB Test | Positive Results | 92 | 1 | 93 | 97.3%<br>( 93.1% ~ 99.3% ) | 96.8%<br>( 90.9% ~ 99.4% ) | 98.1%<br>( 89.5% ~ 100.0% ) | |
| | Negative Results | 3 | 51 | 54 | | | | |
| | Total Results | 95 | 52 | 147 | | | | |
| OC-Light S<br>FIT | Positive Results | 95 | 1 | 96 | 99.3%<br>( 96.2% ~ 100.0% ) | 100.0%<br>( 96.2% ~ 100.0% ) | 98.1%<br>( 89.5% ~ 100.0% ) | |
| | Negative Results | 0 | 51 | 51 | | | | |
| | Total Results | 95 | 52 | 147 | | | | |

#### Polymedco OC Light FOB Test/OC-Light S FIT test results in comparison with the expected results

#### Reaction Time

Reaction time study was conducted in-house with one operator to demonstrate that 5

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Image /page/7/Picture/0 description: The image shows the logo and contact information for Eiken Chemical Co., LTD. The logo is a geometric design of a circle with three points inside. The text includes the company name, address (4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan), and website (http://www.eiken.co.jp).

minutes is the appropriate reaction time for OC-Light S FIT. Hb-free stool specimen was spiked with known levels of hHb to obtain fecal samples with seven different Hb concentrations: 0, 5, 8, 10, 12, 15, and 400 µg/ g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, clinical cutoff), 60 (C95+20%), 75 and 2000 ng/mL. Twenty-one replicates of each sample for a total of 147 samples were tested in a randomized and blinded fashion with results read at 3, 5, 10, and 30 minutes of reaction.

Positivity rate of 25ng/mL was 0%, positivity rate of 40ng/mL was 43%, and positivity rate of 50ng/mL was 100% at 5 minutes of reaction, and the appropriate reaction time of OC-Light S FIT was demonstrated as 5 minutes.

| Reaction time | | Hb concentration (ng/ml) | | | | | | |
|---------------|--------------|--------------------------|------|------|------|------|------|------|
| | | 0 | 25 | 40 | 50 | 60 | 75 | 2000 |
| 3min | Positive | 0 | 0 | 0 | 19 | 21 | 21 | 21 |
| | Negative | 21 | 21 | 21 | 2 | 0 | 0 | 0 |
| | Positive (%) | 0% | 0% | 0% | 90% | 100% | 100% | 100% |
| | Negative (%) | 100% | 100% | 100% | 10% | 0% | 0% | 0% |
| 5min | Positive | 0 | 0 | 9 | 21 | 21 | 21 | 21 |
| | Negative | 21 | 21 | 12 | 0 | 0 | 0 | 0 |
| | Positive (%) | 0% | 0% | 43% | 100% | 100% | 100% | 100% |
| | Negative (%) | 100% | 100% | 57% | 0% | 0% | 0% | 0% |
| 10min | Positive | 0 | 0 | 17 | 21 | 21 | 21 | 21 |
| | Negative | 21 | 21 | 4 | 0 | 0 | 0 | 0 |
| | Positive (%) | 0% | 0% | 81% | 100% | 100% | 100% | 100% |
| | Negative (%) | 100% | 100% | 19% | 0% | 0% | 0% | 0% |
| 30min | Positive | 0 | 2 | 20 | 21 | 21 | 21 | 21 |
| | Negative | 21 | 19 | 1 | 0 | 0 | 0 | 0 |
| | Positive (%) | 0% | 10% | 95% | 100% | 100% | 100% | 100% |
| | Negative (%) | 100% | 90% | 5% | 0% | 0% | 0% | 0% |

Reaction time: test results

#### Precision/Reproducibility

The repeatability and reproducibility of OC-Light S FIT was evaluated by testing twenty-one replicates of fecal samples spiked with hHb to obtain test samples with seven different Hb concentrations: 0, 5, 8, 10, 12, 15, and 400 µg/ g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, clinical cutoff), 60 (C95+20%), 75 and 2000 ng/mL.

The repeatability and reproducibility studies were conducted at three POL sites in the US. The test samples were shipped frozen from our laboratory to the clinical coordinator, where they were collated, and then distributed to the POL sites. There were no cases of invalid or indeterminate results throughout the precision studies.

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Image /page/8/Picture/0 description: The image shows the logo and contact information for Eiken Chemical Co. The logo is a geometric design of a circle with a triangle inside, and three dots are placed at the vertices of the triangle. The text includes the company name, address (4-19-9, Taito, Taito-ku, Tokyo), and website (http://www.eiken.co.jp).

#### u Tokvo 110-8408 http://www.eiken.co.jp

#### Repeatability and reproducibility test results in comparison with the expected

| Type of<br>Precision Study | Actual Results<br>OC-Light S FIT | Expected Results | | | Overall<br>Percent<br>Agreement | Positive Percent<br>Agreement<br>(95% CI) | Negative Percent<br>Agreement<br>(95% CI) |
|-----------------------------------|----------------------------------|---------------------|---------------------|-------|---------------------------------|-------------------------------------------|-------------------------------------------|
| | | Positive<br>Results | Negative<br>Results | Total | | | |
| Repeatability | Positive Results | 95 | 1 | 96 | 99.3% | 100.0%<br>(96.2% - 100.0%) | 98.1%<br>(89.5% - 100.0%) |
| | Negative Results | 0 | 51 | 51 | | | |
| | Total Results | 95 | 52 | 147 | | | |
| Lot-to-Lot<br>Reproducibility | Positive Results | 284 | 6 | 290 | 98.6% | 100.0%<br>(98.7% - 100.0%) | 96.2%<br>(91.8% - 98.6%) |
| | Negative Results | 0 | 151 | 151 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| Between-run<br>Reproducibility | Positive Results | 283 | 0 | 283 | 99.8% | 99.6%<br>(98.0% - 100.0%) | 100.0%<br>(97.7% - 100.0%) |
| | Negative Results | 1 | 157 | 158 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| Between Device<br>Reproducibility | Positive Results | 284 | 6 | 290 | 98.6% | 100.0%<br>(98.7% - 100.0%) | 96.2%<br>(91.8% - 98.6%) |
| | Negative Results | 0 | 151 | 151 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| Between-site<br>Reproducibility | Positive Results | 283 | 6 | 289 | 98.4% | 99.6%<br>(98.0% - 100.0%) | 96.2%<br>(91.8% - 98.6%) |
| | Negative Results | 1 | 151 | 152 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| Combined<br>Reproducibility | Positive Results | 1134 | 18 | 1152 | 98.9% | 99.8%<br>(99.4% - 100.0%) | 97.1%<br>(95.5% - 98.3%) |
| | Negative Results | 2 | 610 | 612 | | | |
| | Total Results | 1136 | 628 | 1764 | | | |

## Test Kit Stability (Accelerated)

Accelerated stability study was conducted with 3 lots of OC-Light S FIT test strips and sampling bottles). The test kit was stored at 60°C for 1.04 months (31 days), 56°C for 1.49 months (45 days), and 50°C for 2.58 months (77 days), which are equivalent to 19 months of storage at 30°C when Arrhenius equation is applied. The test kits stored at above conditions were used to test Hb-free stool specimen spiked with seven different Hb

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Image /page/9/Picture/0 description: The image shows the logo for Eiken Chemical Co., LTD. The logo includes a circular graphic with three dots connected by lines inside a circle. To the right of the graphic is the company name, address (4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan), and website (http://www.eiken.co.jp).

concentrations: 0, 5, 8, 10, 12, 15, and 400 µg/ g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, clinical cutoff), 60 (C95+20%), 75 and 2000 ng/mL.

Overall agreements between the test results and the expected results were statistically over 90% for all of the tests conducted. The results showed that OC-Light S FIT test kit is stable for an estimated period of 19 months at 30°C, enabling claim of 18 months test kit stability at 30°C (room temperature).

| Test kit<br>stability<br>(accelerated) | Actual Results | Expected Results | | | Overall<br>Percent<br>Agreement<br>(95% CI) | Positive<br>Percent<br>Agreement<br>(95% CI) | Negative<br>Percent<br>Agreement<br>(95% CI) |
|----------------------------------------|------------------|---------------------|---------------------|------------------|---------------------------------------------|----------------------------------------------|----------------------------------------------|
| | OC-Light S FIT | Positive<br>Results | Negative<br>Results | Total<br>Results | | | |
| initial | Positive Results | 283 | 2 | 285 | ( 98.0% ~ 99.9% ) | 99.6% | 98.7% |
| | Negative Results | 1 | 155 | 156 | | ( 98.0% ~ 100.0% ) | ( 95.4% ~ 99.9% ) |
| | Total Results | 284 | 157 | 441 | | | |
| 60°C 31 days | Positive Results | 282 | 1 | 283 | ( 98.0% ~ 99.9% ) | 99.3% | 99.4% |
| | Negative Results | 2 | 156 | 158 | | ( 97.4% ~ 99.9% ) | ( 96.4% ~ 100.0% ) |
| | Total Results | 284 | 157 | 441 | | | |
| 56°C 45 days | Positive Results | 282 | 0 | 282 | ( 98.3% ~ 100.0% ) | 99.3% | 100.0% |
| | Negative Results | 2 | 157 | 159 | | ( 97.4% ~ 99.9% ) | ( 97.7% ~ 100.0% ) |
| | Total Results | 284 | 157 | 441 | | | |
| 50°C 77 days | Positive Results | 283 | 3 | 286 | ( 97.7% ~ 99.8% ) | 99.6% | 98.1% |
| | Negative Results | 1 | 154 | 155 | | ( 98.0% ~ 100.0% ) | ( 94.4% ~ 99.6% ) |
| | Total Results | 284 | 157 | 441 | | | |
| | | | | | 99.3% | | |

Test kit stability (accelerated); test results in comparison with the expected results

## Sample Stability

Sample stability study was conducted with three lots of OC-Light S FIT test kit to evaluate the stability of fecal samples collected in OC-Light S FIT sampling bottles. OC-Light S FIT sampling bottles containing fecal samples with seven different Hb concentrations; 0, 5, 8, 10, 12, 15, and 400 µg/ g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, clinical cutoff), 60 (C95+20%), 75 and 2000 ng/mL, were stored at 2, 4, 8, and 10°C for 31 days, and stored at 15, 25, 30, and 32°C for 16 days. The samples stored at 2, 4, 8, and 10°C were tested at day 30 and day 31, and the samples stored at 15, 25, 30, and 32°C were tested at day 15 and day 16, with OC-Light S FIT test strips stored under room temperature.

All analyzed overall agreements between the expected results of OC-Light S FIT utilizing fecal samples in OC-Light S FIT sampling bottles were statistically over 90%. The results showed that the collected fecal samples were stable for 31 days when

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Image /page/10/Picture/0 description: The image shows the logo and contact information for EIKEN CHEMICAL CO., LTD. The logo is a circular design with three interconnected triangles and dots at the vertices. The text includes the company name, address (4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan), and website (http://www.eiken.co.jp).

stored at 2, 4, 8, 10°C, and were stable for 16 days when stored at 15, 25, 30, 32°C, enabling to claim that fecal sample collected in OC-Light S FIT sampling bottle is stable up to 30 days when stored at 2 to 8°C, and is stable up to 15 days when stored at room temperature.

| | Actual Results | | Expected Results | | | | Overall<br>Percent<br>Agreement<br>(95% CI) | | | | Positive<br>Percent<br>Agreement<br>(95% CI) | | | Negative<br>Percent<br>Agreement<br>(95% CI) | | |
|-------------------------------|------------------|--|---------------------|---------------------|------------------|--|---------------------------------------------|--|--|--|----------------------------------------------|--|--|----------------------------------------------|--|--|
| Day 30<br>Sample<br>Stability | OC-Light S FIT | | Positive<br>Results | Negative<br>Results | Total<br>Results | | | | | | | | | | | |
| | Positive Results | | 283 | 6 | 289 | | 98.4%<br>( 96.7% ~ 99.4% ) | | | | 99.6%<br>( 98.0% ~ 100.0% ) | | | 96.2%<br>( 91.8% ~ 98.6% ) | | |
| 2°C | Negative Results | | 1 | 151 | 152 | | | | | | | | | | | |
| | Total Results | | 284 | 157 | 441 | | | | | | | | | | | |
| | | | | | | | | | | | | | | | | |
| | Positive Results | | 284 | 4 | 288 | | 99.1%<br>( 97.7% ~ 99.8% ) | | | | 100.0%<br>( 98.7% ~ 100.0% ) | | | 97.5%<br>( 93.5% ~ 99.3% ) | | |
| 4°C | Negative Results | | 0 | 153 | 153 | | | | | | | | | | | |
| | Total Results | | 284 | 157 | 441 | | | | | | | | | | | |
| | | | | | | | | | | | | | | | | |
| | Positive Results | | 280 | 0 | 280 | | 99.1%<br>( 97.7% ~ 99.8% ) | | | | 98.6%<br>( 96.4% ~ 99.6% ) | | | 100.0%<br>( 97.7% ~ 100.0% ) | | |
| 8°C | Negative Results | | 4 | 157 | 161 | | | | | | | | | | | |
| | Total Results | | 284 | 157 | 441 | | | | | | | | | | | |
| | | | | | | | | | | | | | | | | |
| | Positive Results | | 284 | 2 | 286 | | 99.5%<br>( 98.3% ~ 100.0% ) | | | | 100.0%<br>( 98.7% ~ 100.0% ) | | | 98.7%<br>( 95.4% ~ 99.9% ) | | |
| 10℃ | Negative Results | | 0 | 155 | 155 | | | | | | | | | | | |
| | Total Results | | 284 | 157 | 441 | | | | | | | | | | | |
| | | | | | | | | | | | | | | | | |

#### Sample stability: test results in comparison with the expected results

| | Actual Results | | | | Overall<br>Positive<br>Negative | Percent<br>Agreement | | | | |
|------------------|------------------|----------|------------------|---------|---------------------------------------------------------------|----------------------|--|--|--|--|
| Day 31<br>Sample | OC-Light S FIT | | Expected Results | | Percent<br>Percent<br>Agreement<br>Agreement | | | | | |
| Stability | | Positive | Ne gative | Total | (95% CI)<br>(95% CI)<br>(95% CI) | | | | | |
| | | Results | Results | Results | | | | | | |
| | Positive Results | 283 | 3 | 286 | 99.1%<br>99.6%<br>98.1% | | | | | |
| 2°C | Negative Results | | । ਟੋਕੇ | ારરિ | | | | | | |
| | Total Results | 284 | 157 | 441 | 99.8%<br>~ 99.6%<br>98.0%<br>~ 100.0%<br>97.7%<br>રે<br>94.4% | | | | | |
| | Positive Results | 284 | 2 | 286 | 99.5%<br>100.0%<br>98.7% | | | | | |
| 4°C | Negative Results | 0 | ારરિ | ારર | | | | | | |
| | Total Results | 284 | 157 | 441 | 98.3%<br>~ 100.0%<br>98.7%<br>~ 100.0%<br>95.4%<br>~ 99.9% | | | | | |
| | Positive Results | 283 | 3 | 286 | 99.1%<br>99.6% | | | | | |
| 8°C | Negative Results | | । રત | ારર | | 98.1% | | | | |
| | Total Results | 284 | 157 | 441 | ~ 99.8%<br>~ 99.6%<br>~ 100.0%<br>97.7%<br>98.0%<br>94.4% | | | | | |
| | Positive Results | 280 | 0 | 280 | 99.1%<br>98.6%<br>100.0% | | | | | |
| 10℃ | Negative Results | 4 | 157 | 161 | | | | | | |
| | Total Results | 284 | 157 | 441 | ~ 100.0%<br>97.7%<br>~ 99.8%<br>96.4%<br>~ 99.6%<br>97.7% | | | | | |

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Image /page/11/Picture/0 description: The image shows the logo of EIKEN CHEMICAL CO., LTD. The logo consists of a circular graphic with three dots connected by lines, followed by the company name in bold letters. Below the company name is the address: 4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan, and the website address: http://www.eiken.co.jp.

| | Actual Results | | | | Overall<br>Percent<br>Agreement<br>(95% CI) | Positive<br>Percent<br>Agreement<br>(95% CI) | Negative<br>Percent<br>Agreement<br>(95% CI) |
|-------------------------------|------------------|---------------------|---------------------|------------------|---------------------------------------------|----------------------------------------------|----------------------------------------------|
| Day 15<br>Sample<br>Stability | OC-Light S FIT | Positive<br>Results | Negative<br>Results | Total<br>Results | | | |
| 15°C | Positive Results | 284 | 3 | 287 | 99.3%<br>(98.0% ~ 99.9%) | 100.0%<br>(98.7% ~ 100.0%) | 98.1%<br>(94.4% ~ 99.6%) |
| | Negative Results | 0 | 154 | 154 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| 25°C | Positive Results | 284 | 2 | 286 | 99.5%<br>(98.3% ~ 100.0%) | 100.0%<br>(98.7% ~ 100.0%) | 98.7%<br>(95.4% ~ 99.9%) |
| | Negative Results | 0 | 155 | 155 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| 30°C | Positive Results | 283 | 0 | 283 | 99.8%<br>(98.7% ~ 100.0%) | 99.6%<br>(98.0% ~ 100.0%) | 100.0%<br>(97.7% ~ 100.0%) |
| | Negative Results | 1 | 157 | 158 | | | |
| | Total Results | 284 | 157 | 441 | | | |
| 32°C | Positive Results | 284 | 2 | 286 | 99.5%<br>(98.3% ~ 100.0%) | 100.0%<br>(98.7% ~ 100.0%) | 98.7%<br>(95.4% ~ 99.9%) |
| | Negative Results | 0 | 155 | 155 | | | |
| | Total Results | 284 | 157 | 441 | | | |

| | Actual Results | | Expected Results | | | Overall<br>Percent<br>Agreement<br>(95% CI) | Positive<br>Percent<br>Agreement<br>(95% CI) | Negative<br>Percent<br>Agreement<br>(95% CI) |
|-------------------------------|------------------|-----|---------------------|---------------------|------------------|---------------------------------------------|----------------------------------------------|----------------------------------------------|
| Day 16<br>Sample<br>Stability | OC-Light S FIT | | Positive<br>Results | Negative<br>Results | Total<br>Results | | | |
| 15°C | Positive Results | 284 | 284 | 2 | 286 | 99.5%<br>( 98.3% ~ 100.0% ) | 100.0%<br>( 98.7% ~ 100.0% ) | 98.7%<br>( 95.4% ~ 99.9% ) |
| | Negative Results | 0 | 0 | 155 | 155 | | | |
| | Total Results | 284 | 284 | 157 | 441 | | | |
| 25°C | Positive Results | 283 | 283 | 2 | 285 | 99.3%<br>( 98.0% ~ 99.9% ) | 99.6%<br>( 98.0% ~ 100.0% ) | 98.7%<br>( 95.4% ~ 99.9% ) |
| | Negative Results | 1 | 1 | 155 | 156 | | | |
| | Total Results | 284 | 284 | 157 | 441 | | | |
| 30°C | Positive Results | 284 | 284 | 1 | 285 | 99.8%<br>( 98.7% ~ 100.0% ) | 100.0%<br>( 98.7% ~ 100.0% ) | 99.4%<br>( 96.4% ~ 100.0% ) |
| | Negative Results | 0 | 0 | 156 | 156 | | | |
| | Total Results | 284 | 284 | 157 | 441 | | | |
| 32°C | Positive Results | 284 | 284 | 5 | 289 | 98.9%<br>( 97.4% ~ 99.6% ) | 100.0%<br>( 98.7% ~ 100.0% ) | 96.8%<br>( 92.7% ~ 99.9% ) |
| | Negative Results | 0 | 0 | 152 | 152 | | | |
| | Total Results | 284 | 284 | 157 | 441 | | | |

#### Test Kit Shipping Stress Test

Test kit shipping stress test was conducted to evaluate the effect of exposure to transportation stress during product shipping on OC-Light S FIT test kit. Unused OC-Light S FIT test strips and sampling bottles (containing no fecal samples) were stored under extreme temperature conditions. The measurements were then performed using stressed test strips after fecal samples with seven different Hb concentrations were collected with stressed sampling bottles.

{12}------------------------------------------------

Image /page/12/Picture/0 description: The image shows the logo and contact information for EIKEN CHEMICAL CO., LTD. The logo is a circular design with three interconnected triangles. The text includes the company name, address (4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan), and website address (http://www.eiken.co.jp).

All analyzed overall agreements between the test results of OC-Light S FIT and the expected results were statistically over 90%. The results showed that the test kit was stable for 4 days when stored at -40, -20, 30, 45, and 50°C, enabling to claim that OC-Light S FIT test kit is stable for 3 days of storage at 45°C, and is stable for 3 days of with 3 freeze/thaw cycles.

| Day 2<br>Test kit<br>shipping<br>stress test | Actual Results<br>OC-Light S FIT | Expected Results | | | Overall<br>Percent<br>Agreement<br>(95% CI) | Positive<br>Percent<br>Agreement<br>(95% CI) | Negative<br>Percent<br>Agreement<br>(95% CI) |
|----------------------------------------------|----------------------------------|---------------------|---------------------|------------------|---------------------------------------------|----------------------------------------------|----------------------------------------------|
| | | Positive<br>Results | Negative<br>Results | Total<br>Results | | | |
| -40°C | Positive Results | 283 | 4 | 287 | 98.9% | 99.6% | 97.5% |
| | Negative Results | 1 | 153 | 154 | | | |
| | Total Results | 284 | 157 | 441 | (97.4% ~ 99.6%) | (98.0% ~ 100.0%) | (93.5% ~ 99.3%) |
| -20°C | Positive Results | 284 | 0 | 284 | 100.0% | 100.0% | 100.0% |
| | Negative Results | 0 | 157 | 157 | | | |
| | Total Results | 284 | 157 | 441 | (99.2% ~ 100.0%) | (98.7% ~ 100.0%) | (97.7% ~ 100.0%) |
| 30°C | Positive Results | 283 | 3 | 286 | 99.1% | 99.6% | 98.1% |
| | Negative Results | 1 | 154 | 155 | | | |
| | Total Results | 284 | 157 | 441 | (97.7% ~ 99.8%) | (98.0% ~ 100.0%) | (94.4% ~ 99.6%) |
| 45°C | Positive Results | 284 | 4 | 288 | 99.1% | 100.0% | 97.5% |
| | Negative Results | 0 | 153 | 153 | | | |
| | Total Results | 284 | 157 | 441 | (97.7% ~ 99.8%) | (98.7% ~ 100.0%) | (93.5% ~ 99.3%) |
| 50°C | Positive Results | 282 | 0 | 282 | 99.5% | 99.3% | 100.0% |
| | Negative Results | 2 | 157 | 159 | | | |
| | Total Results | 284 | 157 | 441 | (98.3% ~ 100.0%) | (97.4% ~ 99.9%) | (97.7% ~ 100.0%) |

## Test kit shipping stress test: test results in comparison with the expected results

| Day 3<br>Test kit<br>shipping<br>stress test | Actual Results | Expected Results | | | Overall<br>Percent<br>Agreement<br>(95% CI) | | | Positive<br>Percent<br>Agreement<br>(95% CI) | | | Negative<br>Percent<br>Agreement<br>(95% CI) | | |
|----------------------------------------------|------------------|---------------------|---------------------|------------------|---------------------------------------------|--|--|----------------------------------------------|--|--|----------------------------------------------|--|--|
| | OC-Light S FIT | Positive<br>Results | Negative<br>Results | Total<br>Results | | | | | | | | | |
| -40°C | Positive Results | 283 | 2 | 285 | 99.3% | | | 99.6% | | | 98.7% | | |
| | Negative Results | 1 | 155 | 156 | ( 98.0% ~ 99.9% ) | | | ( 98.0% ~ 100.0% ) | | | ( 95.4% ~ 99.9% ) | | |
| | Total Results | 284 | 157 | 441 | | | | | | | | | |
| -20°C | Positive Results | 284 | 0 | 284 | 100.0% | | | 100.0% | | | 100.0% | | |
| | Negative Results | 0 | 157 | 157 | ( 99.2% ~ 100.0% ) | | | ( 98.7% ~ 100.0% ) | | | ( 97.7% ~ 100.0% ) | | |
| | Total Results | 284 | 157 | 441 | | | | | | | | | |
| 30°C | Positive Results | 284 | 5 | 289 | 98.9% | | | 100.0% | | | 96.8% | | |
| | Negative Results | 0 | 152 | 152 | ( 97.4% ~ 99.6% ) | | | ( 98.7% ~ 100.0% ) | | | ( 92.7% ~ 99.0% ) | | |
| | Total Results | 284 | 157 | 441 | | | | | | | | | |
| 45°C | Positive Results | 284 | 2 | 286 | 99.5% | | | 100.0% | | | 98.7% | | |
| | Negative Results | 0 | 155 | 155 | ( 98.3% ~ 100.0% ) | | | ( 98.7% ~ 100.0% ) | | | ( 95.4% ~ 99.9% ) | | |
| | Total Results | 284 | 157 | 441 | | | | | | | | | |
| 50°C | Positive Results | 279 | 0 | 279 | 98.9% | | | 98.2% | | | 100.0% | | |
| | Negative Results | 5 | 157 | 162 | ( 97.4% ~ 99.6% ) | | | ( 95.9% ~ 99.4% ) | | | ( 97.7% ~ 100.0% ) | | |
| | Total Results | 284 | 157 | 441 | | | | | | | | | |

{13}------------------------------------------------

Image /page/13/Picture/0 description: The image shows the logo for Eiken Chemical Co., LTD. The logo consists of a circular design with three dots inside a triangle, which is enclosed in a circle. Next to the logo is the company name, address (4-19-9, Taito, Taito-ku, Tokyo 110-8408, Japan), and website (http://www.eiken.co.jp).

| Dav 4 | Actual Results | | Expected Results | | | | Overall<br>Percent | | | | | | Positive<br>Percent | Negative<br>Percent | | | | | |
|----------------------|------------------|----------|---------------------|------------------|-------------------------------|----------------------|--------------------…