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

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

A. 510(k) Number:
K143325

B. Purpose for Submission:
Clearance of a new device

C. Measurand:
Human hemoglobin (hHb) in feces

D. Type of Test:
Lateral flow chromatographic immunoassay

E. Applicant:
EIKEN CHEMICAL CO., LTD

F. Proprietary and Established Names:
OC-Light S FIT

G. Regulatory Information:

1. Regulation section:
21 CFR 864.6550, Occult blood test

2. Classification:
Class II

3. Product code:
KHE, Reagent, Occult Blood

4. Panel:
Hematology (81)

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H. Intended Use:

1. Intended use(s):

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.

2. Indication(s) for use:

Same as Intended Use

3. Special conditions for use statement(s):

For prescription use only

4. Special instrument requirements:

Not applicable

I. Device Description:

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

1. Components of OC-Light S FIT: OC-Light S FIT consists of a test strip and a sampling bottle.

a. 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.

b. OC-Light S FIT sampling bottle is a plastic bottle for collecting fecal sample. It contains 2.0 mL of extraction buffer. The sampling bottle is specially designed for:

i. Proper sampling of feces: Sample probe is attached to the top bottle cap. The sample probe insertion slot is designed to rub off excess amount of stool before it reaches the area with extraction buffer, enabling only stool caught in the grooved portion (approximately 10 mg) of the sample probe to be mixed with the buffer solution.

ii. Thorough mixing of fecal sample with extraction buffer: Sample probe itself acts as a stopper, preventing excess amount of collected stool from entering into the extraction buffer area and buffer solution from leaking outside.

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J. Substantial Equivalence Information:

1. Predicate device name(s):
Polymedco OC-Light FOB Test

2. Predicate 510(k) number(s):
K041297

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device OC-Light S FIT, K143325 | Predicate Polymedco OC-Light FOB Test, K041297 |
| Intended Use | 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. | An immunological test intended for the detection of fecal occult blood in feces by professional laboratories and physician office labs. The test is useful for the determination of gastrointestinal (GI) bleeding, found in a number of gastrointestinal (GI) disorders, e.g., diverticulitis, colitis, polyps, and colorectal cancer. |
| Indications for Use | Recommended for use in routine physical examinations. | Recommended for use in 1) routine physical examinations, 2) monitoring for bleeding in patients, and 3) screening for colorectal cancer or gastrointestinal bleeding. |
| Sample Type | Feces in an extraction buffer | Feces in an extraction buffer |
| Test Principle | Qualitative test system intended for immunochemical detection of fecal occult blood in feces. | Immunochemical test system intended for qualitative detection of fecal occult blood in feces. |
| Detection Method | Lateral flow chromatographic immunoassay | Lateral flow chromatographic immunoassay |
| Assay Cut-off | 10 μg hemoglobin/g stool or 50 ng/mL buffer (Human hemoglobin in human fecal sample mixed with detection buffer) | 50 ng/mL (Human hemoglobin in human fecal sample mixed with detection buffer) |

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| Differences | | |
| --- | --- | --- |
| Item | Device
OC-Light S FIT, K143325 | Predicate
Polymedco OC-Light FOB Test,
K041297 |
| 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 |

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

CLSI EP25-A Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline-Second Edition, 2009.

CLSI EP12-A2 User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline-Second Edition, 2008.

CLSI EP05-A3 Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline-Third Edition.

## L. Test Principle:

OC-Light S FIT is a qualitative test designed for the immunochemical detection of human hemoglobin (hHb) in stool specimens. When the sample end of 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 visible reddish/pink line. The procedural control region of the membrane contains anti-mouse antibodies that capture any unreacted/excess antibody-gold conjugates, forming a distinct reddish/pink line. The reddish/pink line in the procedural control region demonstrates the validity of the test.

## M. Performance Characteristics:

### 1. Analytical performance:

#### a. Precision/Reproducibility:

The repeatability of OC-Light S FIT was evaluated at one intended use site, using one test kit lot and one operator. The repeatability study was performed using 21 replicates of fecal test samples collected in OC-Light S FIT sampling bottles from Hb-free stool specimens spiked with human blood to obtain test samples containing seven different concentrations of human hemoglobin: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cutoff), 60 (C95+20%), 75, and 2000 ng/mL, respectively.

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Reproducibility studies were conducted at three intended use sites in the U.S. The reproducibility of OC-Light S FIT was evaluated by testing 21 replicates of fecal test samples collected in OC-Light S FIT sampling bottles from Hb-free stool specimens spiked with human blood to obtain test samples containing seven different concentrations of human hemoglobin: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75, and 2000 ng/mL, respectively.

OC-Light S FIT repeatability and reproducibility testing was carried out in a random and blinded manner. No invalid or indeterminate results were obtained throughout the precision studies. FOBT-CHECK 'Negative and Positive Controls' were also tested daily to ensure and confirm the validity of the test results.

Statistical analysis of repeatability and reproducibility studies of OC-Light S FIT

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

b. Linearity/assay reportable range:

Prozone (Hook Effect)

Susceptibility of OC-Light S FIT to prozone effect was evaluated by testing Hb-free stool specimens spiked with Human HbA0 solution, HbS whole blood and HbC whole blood. Test samples were prepared by spiking Hb-free stool specimen with known levels of human HbA0 solution to obtain fecal samples with eight different Hb concentrations of each: 0 μg/g stool, 20 μg/g stool, 100 μg/g stool, 150 μg/g stool, 200 μg/g stool, 250 μg/g stool, 300 μg/g stool, and 400 μg/g stool, that are equivalent to 0, 100, 500, 750, 1000, 1250, 1500, and 2000 ng/mL, respectively. Twenty one aliquots of each sample

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concentration were mixed with extraction buffer and tested in randomized order.

Test samples were also prepared by spiking Hb-free stool specimens with known levels of HbS supernatant to obtain fecal samples with eight different HbS concentrations and known levels of HbC supernatant to obtain fecal samples with eight different HbC concentrations: 0 µg/g stool, 20 µg/g stool, 100 µg/g stool, 150 µg/g stool, 200 µg/g stool, 250 µg/g stool, 300 µg/g stool, and 400 µg/g stool, that are equivalent to 0, 100, 500, 750, 1000, 1250, 1500, and 2000 ng/mL, respectively. Twenty one aliquots of each sample concentration were mixed with extraction buffer and tested in randomized order.

FOBT-CHECK ‘Negative and Positive Controls’ were also tested to ensure and confirm the validity of the test results of the test prozone/hook effect study. OC-Light S FIT was not found susceptible to prozone/hook effect as it would display positive test results with fecal samples containing a hemoglobin concentration up to 2000 ng/mL.

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

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

External Control: Controls are not provided with this kit. It is recommended that positive and negative controls be performed to verify proper test performance. The FOBT-CHECK Control Kit, cleared under K041297, is recommended for use as the external controls.

Stability Studies

1. Stability of fecal samples

Samples for stability testing were prepared by spiking stool samples with human blood (of known hemoglobin level) to obtain fecal samples containing seven different hemoglobin concentrations: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Twenty one aliquots of each of the seven concentrations of spiked stool test samples were mixed with detection buffer in OC-Light S FIT sample collection bottles and analyzed in random order.

To claim 30 days of sample stability at 2 to 8°C each sample was tested with OC-Light S FIT and stored at 2, 4, 8, and 10°C at time points of 15, 30, and 31 days from the start of storage. OC-Light S FIT sampling bottles must be brought back to room temperature before testing.

To claim 15 days of sample stability at room temperature each sample was tested with OC-Light S FIT and stored at 15, 25, 30, and 32°C at time points of 7, 15, and 16 days from the start of storage.

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FOBT-CHECK ‘Negative and Positive Controls’ were also tested daily to ensure and confirm the validity of the test results of the stability study. Stool samples collected in OC-Light S FIT sampling bottles are stable up to 30 days when stored at 2 to 8°C, and stable up to 15 days when stored at room temperature.

2. Test Kit Stability (Accelerated)

Accelerated stability study was conducted with 3 lots of OC-Light S FIT test kit (test strips and sampling bottles). Test kit stability testing was performed by preparing 21 aliquots of spiked stool test samples collected in detection buffer in OC-Light S FIT sample collection bottles. The Hb-free stool specimens were spiked with human blood (of known hemoglobin level) to obtain fecal samples containing seven different hemoglobin concentrations: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Each spiked stool test sample was analyzed in random order. The three lots of OC-Light S FIT test strips (contained in canisters) and sampling bottles were stored and tested at 50°C, 56°C and 60°C. The test kit stability was performed at the following time intervals:

Device kit stored at 60°C: 1.04 months (31 days) from the start of storage.

Device kit stored at 56°C: 1.49 months (45 days) from the start of storage.

Device kit stored at 50°C: 2.58 months (77 days) from the start of storage.

FOBT-CHECK ‘Negative and Positive Controls’ were also tested daily to ensure and confirm the validity of the test results of the stability study. The results showed that the test kit was stable for an estimated period of 18 months at 30°C when Arrhenius equation is applied.

3. Fecal Sample Shipping Stress Test

Three sets of OC-Light S FIT test kits (3 lots of test strips plus 3 lots of sampling bottles) were evaluated during the fecal sample shipping stress test. Fecal samples were prepared by spiking stool samples with human blood (of known hemoglobin level) to obtain fecal samples containing seven different hemoglobin concentrations: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Twenty one aliquots of each of the seven concentrations of spiked stool test samples were mixed with detection buffer in OC-Light S FIT sample collection bottles. The initial tests were performed with randomized samples that are not stressed to confirm that all sets give expected results. The OC-Light S FIT sampling bottles with fecal samples of known Hb concentrations were stored at -40°C, -20°C, 30°C, 45°C, and 50°C. The stool samples/test kit stored at -40°C and -20°C were brought to room temperature on day 1 to have a freeze-thaw cycle, and was then returned to storage. At time points of 2, 3, and 4 days from the start of storage, 147 OC-Light S FIT sampling bottles (21 aliquots of 7 Hb concentrations) of each set from the storage were taken out and brought back to room temperature. The samples were randomized and tested with a paired lot of OC-Light S FIT test strips. FOBT-CHECK ‘Negative and Positive

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Controls' were also tested daily to ensure and confirm the validity of the test results of the stability study. Studies demonstrated that fecal samples collected in OC-Light S FIT sampling bottles are stable for 3 days of storage at 45°C and stable for 3 days with 3 freeze/thaw cycles.

## 4. Test Kit Shipping Stress Test

Three sets of OC-Light S FIT test kits (3 lots of test strips plus 3 lots of sampling bottles) were evaluated during the test kit shipping stress test. The unused sets of OC-Light S FIT test strips and sampling bottles (containing no fecal samples) were stored at -40°C, -20°C, 30°C, 45°C, and 50°C. The test kit stored at -40°C and -20°C were brought to room temperature on day 1 to have a freeze-thaw cycle, and was then returned to storage. At time points 2, 3, and 4 days from the start of storage, 147 OC-Light S FIT test strips and sampling bottles of each test set from the storage were taken out and brought back to room temperature. To evaluate the performance of stressed test kits, fecal samples for testing were prepared by spiking stool samples with human blood (of known hemoglobin level) to obtain fecal samples containing seven different hemoglobin concentrations: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL. Twenty one aliquots of each of the seven concentrations of spiked stool test samples were mixed with detection buffer in OC-Light S FIT sample collection bottles. The samples were randomized and tested with a paired lot of OC-Light S FIT test strips. FOBT-CHECK 'Negative and Positive Controls' were also tested daily to ensure and confirm the validity of the test results of the stability study. Studies demonstrated that OC-Light S FIT test kits are stable for 3 days of storage at 45°C and stable for 3 days with 3 freeze/thaw cycles.

## d. Detection limit:

Not applicable

## e. Analytical specificity:

### Specificity to human hemoglobin variant

The ability of OC-Light S FIT to detect human hemoglobin variants was determined by testing hemoglobin-S (HbS) and hemoglobin-C (HbC) by preparing spiked stool samples containing seven different hemoglobin concentrations: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Twenty one aliquots of each of the seven concentrations (7 HbS and 7 HbC supernatants) of spiked stool test samples were mixed with detection buffer in OC-Light S FIT sample collection bottles. Samples were tested in a randomized order. Results showed OC-Light S FIT equivalently recognizes variants of hemoglobin HbS and HbC.

### Cross-Reactivity

Cross-reactivity of OC-Light S FIT with animal hemoglobin was evaluated by testing

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prepared test samples in OC-Light S FIT sample collection bottles by spiking Hb-free stool specimen with known levels of human Hb solution to obtain fecal samples with seven different Hb concentrations: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool, that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Twenty one replicates of spiked hemoglobin-negative stool concentrations listed above were then spiked with the intended level of respective animal hemoglobin such as bovine (500 μg/mL), turkey (500 μg/mL), fish (500 μg/mL), equine (500 μg/mL), goat (500 μg/mL), rabbit (500 μg/mL), porcine (500 μg/mL) and sheep (500 μg/mL). All samples were tested in a randomized order. OC-Light S FIT did not show significant cross-reactivity with any of the animal hemoglobins tested.

## Interfering Substances

Susceptibility of OC-Light S FIT to interference from extracts (animal meat and vegetable) was evaluated by testing 21 replicates of prepared test samples by spiking Hb-free stool specimen with known levels of human Hb solution to obtain fecal samples with seven different Hb concentrations: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. The 21 replicates of the spiked hemoglobin-negative stool concentrations listed above were spiked with the intended level of respective animal meat extracts: beef meat extract (2.5%), pork meat extract (2.5%), fish meat extract (2.5%), horse meat extract (2.5%), goat meat extract (2.5%), rabbit meat extract (2.5%), lamb meat extract (2.5%), and chicken meat extract (2.5%).

The 21 replicates of hemoglobin-negative stool spiked listed above were also spiked with the intended level of respective vegetable extracts: broccoli extracts (2.5%), cantaloupe extract (2.5%), cauliflower extract (2.5%), horseradish extract (2.5%), parsnip extract (2.5%), red radish extract (2.5%), and turnip extract (2.5%). All samples were mixed with detection buffer in OC-Light S FIT sample collection bottles and tested in a randomized order. OC-Light S FIT did not show significant interference from any of the animal meat or vegetable extracts.

## Interfering Supplements

Susceptibility of OC-Light S FIT to interference from iron and sodium L-ascorbate was evaluated by testing 21 replicates of prepared test samples by spiking Hb-free stool specimen with known levels of human Hb solution to obtain fecal samples with seven different Hb concentrations: 0 μg/g stool, 5 μg/g stool, 8 μg/g stool, 10 μg/g stool, 12 μg/g stool, 15 μg/g stool, and 400 μg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. The 21 replicates of spiked hemoglobin-negative stool concentrations were spiked with iron (0.5% (w/v)), and sodium L-ascorbate (0.5% (w/v)). All samples were mixed with detection buffer in OC-Light S FIT sample collection bottles and tested in a randomized order. OC-Light S FIT did not show significant interference from iron and sodium L-ascorbate.

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# Interference from Toilet Cleaners

Susceptibility OC-Light S FIT to interference from toilet cleaners was evaluated by testing 21 replicates of prepared test samples by spiking Hb-free stool specimen with known levels of human Hb solution to obtain fecal samples with seven different Hb concentrations: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL respectively. The 21 replicates of spiked hemoglobin-negative stool concentrations were spiked with the following cleaners: ef Bio (5 mg/mL), LIME A-WAY (5 mg/mL), Nuriper (5 mg/mL), Lysol bleach (5 mg/mL), Lysol cleaner (5 mg/mL), Scrubbing Bubbles (5 mg/mL). All samples were mixed with detection buffer in OC-Light S FIT sample collection bottles and tested in a randomized order. OC-Light S FIT did not show significant interference from substances contained in the tested toilet cleaners.

FOBT-CHECK ‘Negative and Positive Controls’ were also tested to ensure and confirm the validity of the test results of the interference studies.

## f. Assay cut-off:

Fecal test samples for cut-off testing were prepared by spiking stool samples with human blood (of known hemoglobin level) to obtain fecal samples containing seven different hemoglobin concentrations: 0 µg/g stool, 5 µg/g stool, 8 µg/g stool, 10 µg/g stool, 12 µg/g stool, 15 µg/g stool, and 400 µg/g stool that are equivalent to 0, 25 (C5), 40 (C50, C95-20%), 50 (C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL, respectively. Twenty one aliquots of each of the seven concentrations of spiked stool test samples were mixed with detection buffer in OC-Light S FIT sample collection bottles and Polymedco OC-Light FOB Test sample bottles. Samples were tested in randomized order. Testing was performed side-by-side with the predicate by comparing the test results of the device with that of the predicate. Cut-off was determined to be 10 µg hemoglobin/g stool or 50 ng/mL (hemoglobin in fecal sample mixed with detection buffer).

### Polymedco OC-Light FOB/OC-Light S FIT test results versus expected results

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

## 2. Comparison studies:

a. Method comparison with predicate device:

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A method comparison of OC-Light S FIT with the predicate test, Polymedco OC-Light FOB Test, was conducted by assessing patient samples and one Hb-spiked sample of known concentration at $75\mathrm{ng / mL}$ (near the cut-off) at each intended use site, physician office laboratories (POLs) and professional medical laboratories (PMLs). The FOBT-CHEK external controls (positive and negative) were run prior to testing. Statistical analysis of site-wise test results as well as combined results showed that OC-Light S FIT test results have acceptable overall percent agreement as well as positive percent agreement and negative percent agreement with Polymedco OC-Light FOB test results.

Method Comparison study test results

| POLs Method Comparison Study | New Test | Predicate Test | | | Overall Percent Agreement (95% CI) | Positive Percent Agreement (95% CI) | Negative Percent Agreement (95% CI) |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | OC-Light S FIT | Polymedco OC-Light FOB TEST | | | | | |
| | | Positive Results | Negative Results | Total Results | | | |
| POL 1 | Positive Results | 17 | 0 | 17 | 100.0% (96.6%~100.0%) | 100.0% (83.8%~100.0%) | 100.0% (96.0%~100.0%) |
| | Negative Results | 0 | 90 | 90 | | | |
| | Total Results | 17 | 90 | 107 | | | |
| POL 2 | Positive Results | 13 | 1 | 14 | 99.1% (94.9%~100.0%) | 100.0% (79.4%~100.0%) | 99.0% (94.2%~100.0%) |
| | Negative Results | 0 | 95 | 95 | | | |
| | Total Results | 13 | 96 | 109 | | | |
| POL 3 | Positive Results | 15 | 0 | 15 | 100.0% (96.4%~100.0%) | 100.0% (81.8%~100.0%) | 100.0% (95.8%~100.0%) |
| | Negative Results | 0 | 85 | 85 | | | |
| | Total Results | 15 | 85 | 100 | | | |
| POLs combined | Positive Results | 45 | 1 | 46 | 99.7% (98.2%~100.0%) | 100.0% (92.1%~100.0%) | 99.6% (97.9%~100.0%) |
| | Negative Results | 0 | 270 | 270 | | | |
| | Total Results | 45 | 271 | 316 | | | |
| PMLs Method Comparison Study | New Test | Predicate Test | | | Overall Percent Agreement (95% CI) | Positive Percent Agreement (95% CI) | Negative Percent Agreement (95% CI) |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | OC-Light S FIT | Polymedco OC-Light FOB TEST | | | | | |
| | | Positive Results | Negative Results | Total Results | | | |
| PML 1 | Positive Results | 21 | 0 | 21 | 100.0% (98.3%~100.0%) | 100.0% (86.7%~100.0%) | 100.0% (98.1%~100.0%) |
| | Negative Results | 0 | 194 | 194 | | | |
| | Total Results | 21 | 194 | 215 | | | |
| PML 2 | Positive Results | 18 | 0 | 18 | 100.0% (98.3%~100.0%) | 100.0% (84.6%~100.0%) | 100.0% (98.1%~100.0%) |
| | Negative Results | 0 | 193 | 193 | | | |
| | Total Results | 18 | 193 | 211 | | | |
| PML 3 | Positive Results | 37 | 0 | 37 | 100.0% (98.3%~100.0%) | 100.0% (90.5%~100.0%) | 100.0% (97.9%~100.0%) |
| | Negative Results | 0 | 174 | 174 | | | |
| | Total Results | 37 | 174 | 211 | | | |
| PMLs combined | Positive Results | 76 | 0 | 76 | 100.0% (99.4%~100.0%) | 100.0% (95.3%~100.0%) | 100.0% (99.3%~100.0%) |
| | Negative Results | 0 | 561 | 561 | | | |
| | Total Results | 76 | 561 | 637 | | | |

The above method comparison study demonstrated that the analytical performance of the OC-Light S FIT test is substantially equivalent to the predicate device.

# b. Matrix comparison:

Not applicable

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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. Instrument Name:
Not applicable

O. System Descriptions:

1. Modes of Operation:
Does the applicant’s device contain the ability to transmit data to a computer, webserver, or mobile device?
Yes _______ or No ☐ x _______

Does the applicant’s device transmit data to a computer, webserver, or mobile device using wireless transmission?
Yes _______ or No ☐ x _______

2. Software:
FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:
Yes _______ or No ☐ x _______

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3. Specimen Identification:

Enter patient identification information manually on the label of the Sampling Bottle.

4. Specimen Sampling and Handling:

It is recommended that the laboratory should give the OC-Light S FIT Sample Collection Bottle to the patient so that the patient can collect the fecal sample and do the sampling himself/herself and submit it to the laboratory for testing as per the appropriate instructions received from the laboratory. The sample must be returned or mailed to the doctor/laboratory within 5 days of sampling. The laboratory may store sample at room temperature for up to 15 days or can be refrigerated at 2–8°C (36–46°F) for up to 30 days.

5. Calibration:

Not applicable

6. Quality Control:

Internal Control: The Procedural Control is found in the Procedural Control Region of the Test Strip to assure the operator that (a) sample addition and migration through the test strip has occurred and (b) the control anti-mouse antibody and the reporter MAb are intact and functional. This control does not ensure that the capture antibody is accurately detecting the presence or absence of Hb in the sample.

External controls: External controls are used to assure the operator that the capture and conjugated antibodies are present and reactive. Controls should be assayed according to manufacturer instructions once per kit lot, following the local and state guidelines. If controls do not perform as expected, the test results should not be used.

P. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

1. Test Kit Reaction Time:

Reaction time test samples were prepared by spiking Hb-free stool specimen with known levels of human Hb solution to obtain fecal samples with seven different Hb concentrations: 0, 25 (C5), 40 (C50, C95-20%), 50(C95, cut-off), 60 (C95+20%), 75 and 2000 ng/mL. Each sample was collected with 21 OC-Light S FIT sampling bottles and tested in a randomized order. The test was performed in-house with OC-Light S FIT test strips and the results read at 3, 5, 10, and 30 minutes. 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. The appropriate reaction time of OC-Light S FIT was demonstrated as 5 minutes.

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2. Operator Intensity Reading:

The operator intensity reading test was performed by utilizing seven (7) levels of stool samples spiked with Hb concentrations by adding human Hb to Hb-free stool samples with concentrations at 0, 25, 37.5 (C95-25%), 50 (C95, cut-off), 62.5(C95 +25%), 75, and 2000 ng/mL. The negative and positive controls were performed in the same manner as the spiked stool test samples. Five groups of samples were created, each group consisting of seven samples per concentration and controls each tested in a randomized order. Five participant readers from an intended use site analyzed the group of samples and recorded the results and graded the intensity of the test line from 6 levels: N (negative), F (faint), 1+, 2+, 3+, and 4+. The intensity of the test line increased in correlation with the Hb concentration of stool test samples. There was no statistical significance in the analyses of the test samples performed by the operators for the intensity reading test study.

Q. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

R. Conclusion:

The submitted information in this premarket notification is complete and supports substantial equivalence decision.