INDICAID PI-0002 COVID-19 Rapid Antigen At Home Test Instruction Manual

INDICAID PI-0002 COVID-19 Rapid Antigen At Home Test Instruction Manual

Intended Use

The INDICAID™ COVID-19 Rapid Antigen At-Home Test is a rapid lateral flow immunoassay device intended for the qualitative detection of nucleocapsid protein antigen from SARS-CoV-2. This test is authorized for non-prescription home use with self-collected anterior nasal (nares) swab samples from individuals aged 14 years or older, or adult-collected anterior nasal (nares) swab samples from individuals aged 2 years or older. This test is authorized for individuals with symptoms of COVID-19 within the first 6 days of symptom onset when tested at least twice over three days with at least 48 hours between tests, and for individuals without symptoms or other epidemiological reasons to suspect COVID-19, when tested at least three times over five days with at least 48 hours between tests.

The INDICAID™ COVID-19 Rapid Antigen At-Home Test does not differentiate between SARS-CoV and SARS-CoV-2.

Results are for the identification of SARS-CoV-2 nucleocapsid protein antigen. Antigen is generally detectable in anterior nasal (nares) samples during the acute phase of infection. Positive results indicate the presence of viral antigens, but clinical correlation with past medical history and other diagnostic information is necessary to determine infection status. Positive results do not rule out bacterial infection or co-infection with other viruses and the agent detected may not be the definite cause of disease. Individuals who test positive with the INDICAID™ COVID-19 Rapid Antigen At-Home Test should self-isolate and seek follow-up care with their physician or healthcare provider as additional testing may be necessary.

All negative results are presumptive and confirmation with a molecular assay, if necessary, for patient management, may be performed. Negative results do not rule out SARS-CoV-2 infection and should not be used as the sole basis for treatment or patient management decisions, including infection control measures, such as isolating from others and wearing masks.

Negative results should be considered in the context of a patient’s recent exposures, history, and the presence of clinical signs and symptoms consistent with COVID-19. Individuals who test negative and continue to experience COVID-19 like symptoms of fever, cough, and/or shortness of breath may still have a SARS-CoV-2 infection and should seek follow up care with their healthcare provider.

Individuals should provide all results obtained with this product to their healthcare provider for public health reporting and to receive appropriate medical care. All healthcare providers will report all test results they receive from individuals who use the authorized product to relevant public health authorities in accordance with local, state, and federal requirements using appropriate LOINC and SNOMED codes, as defined by the Laboratory In Vitro Diagnostics (LIVD) Test Code Mapping for SARSCoV-2 Tests provided by CDC.
The INDICAID™ COVID-19 Rapid Antigen At-Home Test is intended for non- prescription self-use and/or as applicable, for an adult lay user testing another aged 2 years or older in a non-laboratory setting. The INDICAID™ COVID-19 Rapid Antigen At-Home Test is only for in vitro diagnostic use under the Food and Drug Administration’s Emergency Use Authorization. This product has not been FDA cleared or approved.

Explanation of the Test

COVID-19 (short for “Coronavirus disease 2019”) is a disease first recognized in 2019 that is caused by type of novel coronavirus called SARS-CoV-2. Due to its rapid spread,
the World Health Organization (WHO) recognized the disease as a global pandemic on March 11, 2020. Many individuals with confirmed COVID-19 have developed fever and/or symptoms of acute respiratory illness (e.g., cough, dyspnea), although some individuals experience only mild symptoms or no symptoms at all. The virus is spread primarily from person to person through respiratory particles, even by individuals without symptoms.

The INDICAID™ COVID-19 Rapid Antigen At-Home Test is a rapid qualitative immunochromatographic assay for the determination of the presence of SARS- CoV-2 antigens in anterior nasal swab specimens. Each INDICAID™ COVID-19 Rapid Antigen At-Home Test is single-use and can analyze one anterior nasal (nares) swab sample. The total time required to perform one test is approximately 20 minutes from clinical specimen collection to result.

SARS-CoV-2-specific antibodies and a control antibody are immobilized onto a nitrocellulose membrane support as two distinct lines. The test line (T) region contains monoclonal anti-SARS-CoV-2 antibodies and the control line (C) regioncontains polyclonal control antibodies. Polyclonal and monoclonal anti- SARS-CoV-2 antibodies conjugated with red-colored latex microspheres are used to detect the SARS-CoV-2 antigen.

During the test, the swab containing patient sample is placed and mixed in a buffer solution vial. That buffer solution is then applied to the sample well of the test device. If SARS-CoV-2 antigen is present, it will bind to the antibody-latex microsphere conjugate forming an immunocomplex. The immunocomplex will then travel across the strip via capillary action towards the test line. The immunocomplex will then bind to the anti-SARS-CoV-2 antibodies at the test line (T), forming a visible red-colored line to indicate detection of antigens. If SARS-CoV-2 antigens are not detected in the sample, no color will appear at the test line (T). Test results are interpreted visually at 20 minutes after the sample has been properly applied to the test according to the instructions. Results should not be read after 25 minutes.

The control (C) line is used for procedural control and should appear regardless of the test result. The appearance of the control line (C) serves to ensure the test is performing properly and the test result is valid.

The INDICAID™ COVID-19 Rapid Antigen At-Home Test is validated for use from direct specimens testing without transport media.

Materials Provided

containing one test strip in a plastic device cassette. Each strip has one control line and one test li
Buffer solution vials| 2, 4, 12, or 24| Vial with cap and integrated dispensing tip, containing 400 μL of buffer solution
Nasal swabs| 2, 4, 12, or 24| Individually wrapped, sterile specimen collector.
Package insert| 1 User Instructions/ Quick Reference Guide| Documentation for user instructions.

MATERIALS REQUIRED BUT NOT PROVIDED

• Timer

Warnings, Precautions, and Safety Information

• Read all instructions carefully before performing test. Failure to follow directions may result in inaccurate test results.

• In the USA, this product has not been FDA cleared or approved, but has been authorized by FDA under an Emergency Use Authorization (EUA). This product is only authorized for the detection of proteins from SARS-CoV-2,
  not for any other viruses or pathogens. The emergency use of this product is only authorized for the duration of the declaration that circumstances exist justifying the authorization of emergency use of in vitro diagnostics for detection and/or diagnosis of COVID-19 under Section 564(b)(1) of the Act, 21
  U.S.C. § 360bbb-3(b)(1), unless the declaration is terminated or authorization is revoked sooner.

• Serial testing should be performed in individuals with negative results at least twice over three days (with 48 hours between tests) for symptomatic individuals and three times over five days (with at least 48 hours between tests) for asymptomatic individuals. You may need to purchase additional tests to perform this serial (repeat) testing.

• An anterior nasal swab sample can be self-collected by an individual aged 14 years and older. Children aged 2 to 13 years should be tested by an adult.

• Do not use the test on children under 2 years of age.

• Do not eat, drink, or smoke in the area where the specimens and kit contents are handled.

• Wear a safety mask or other face-covering when collecting a specimen from a child or another individual.

• Do not use if any of the test kit contents or packaging is damaged.

• All test components are single use. Do not re-use.

• Do not use this test kit beyond the expiration date printed on the outside of the box.

• Use only the contents provided in the test kit.

• Do not mix components from different kit lots.

• Wash hands thoroughly for at least 20 seconds before and after using the test.

• Leave the swab inside its packaging until instructed to swab the nose. Keep the swab clean. Do not allow anything to touch the soft tip of the swab until instructed to swab the nose.

• When collecting a sample, use only the nasal swab provided in the kit.

• Once opened, the test device should be used within 2 hours.

• Perform the test as soon as possible after swabbing both nostrils, and within 30 minutes after adding the swab to the vial.

• This test is read visually. User with impaired vision or color impaired vision may
  not be able to read the test.

• Do not read the test result before 20 minutes or after 25 minutes, following application of the sample to the test device. Results read before 20 minutes or after 25 minutes may lead to a false positive, false negative, or invalid result.

• If you have had symptoms longer than 6 days you should consider testing at least Three times over five days with at least 48 hours between tests.

• False negative results may occur if insufficient buffer solution is applied to the test device (e.g. less than 3 drops).

• False negative results may occur if the swab is not twisted 20 times in the buffer solution vial. False negative results may occur if the swab head is not rolled against the inner wall of the buffer solution vial to release as much liquid from the swab as possible.

• Keep foreign substances and household cleaning products away from the test        during the testing process. Contact with foreign substances and household cleaning products (e.g., 1% bleach) may result in an incorrect test result.

• Test devices that contain patient samples should be handled as though they could transmit disease. Follow universal precautions when handling samples, this kit, and its contents. Wear appropriate personal protection equipment (PPE) and gloves when running the test and handling a patient’s test device. Change gloves between tests.

• Dispose of used specimens and test components in accordance with Federal, State, and Local requirements.

• In the event of a spillage, ensure it is cleaned thoroughly using a suitable disinfectant.

• Keep testing kit and all test kit components out of the reach of children and pets before and after use. Avoid exposure of your skin, eyes, nose, or mouthto the solution in the tube. Do not ingest any kit components. The reagent solution contains harmful chemicals (see table below). If the solution contacts your skin, eyes, nose, or mouth, flush with large amounts of water. If irritation persists, seek medical advice: https://www.poisonhelp.org or 1-800-222-1222.

• For additional information on hazard symbols, safety, handling and disposal of the components within this kit, please refer to the Safety Data Sheet (SDS) located at us.phasescientific.com.

• For more information on EUAs please visit: https://www.fda.gov/– emergencypreparedness-and-response/mcm-legal-regulatory-and policyframework/emergencyuse-authorization

• For the most up to date information on COVID-19, please visit: www.cdc.gov/COVID19

and eye damage H317, May cause an allergic skin reaction H410, Toxic to aquatic life| 0.3% v/v1

Limitations

• The performance of this test was established based on the evaluation of a limited number of clinical specimens collected between December 2021 and January 2022. The clinical performance has not been established in all circulating variants but is anticipated to be reflective of the prevalent variants in circulation at the time and location of the clinical evaluation. Performance at the time of testing may vary depending on the variants circulating, including newly emerging strains of SARS-CoV-2 and their prevalence, which change over time.
• There is a higher chance of false negative results with antigen tests than with laboratory-based molecular tests due to the sensitivity of the test technology. This means that there is a higher chance this test will give a false negative result in an individual with COVID-19 as compared to a molecular test, especially in samples with a low viral load.
• All COVID-19 antigen test negative results are presumptive and confirmation with a molecular assay may be necessary.
• If the patient continues to have symptoms of COVID-19, and both the patient’s first and second tests are negative, the patient may not have COVID-19, however additional follow-up may be needed.
• If the test is positive, then proteins from the virus that causes COVID-19 have been found in the sample and the individual likely has COVID-19.
• This test is read visually and has not been validated for use by those with impaired vision or color-impaired vision.
• Incorrect test results may occur if a specimen is incorrectly collected or handled.
• This test detects both viable (live) and nonviable SARS-CoV-2. Test performance depends on the amount of virus (antigens) in the sample and may or may not correlate with viral culture results performed on the same sample.

Storage and Stability

• The INDICAID™ COVID-19 Rapid Antigen At-Home Test should be stored in a cool, dry place between 2-30°C (35.6-86°F). Do not freeze. Avoid direct sunlight.
• Kit components in the INDICAID™ COVID-19 Rapid Antigen At-Home Test are stable until the expiration date printed on the label. For information about current expiration dates for at-home OTC COVID-19 diagnostic tests, visit http://www.fda.gov/covid-tests.
• The test device must remain in the sealed foil pouch until use. Once the pouch has been opened, the test device should be used within 60 minutes.
• Test samples immediately after collection, but no more than 5 minutes after specimen collection before placement into extraction buffer or up to 2 hours after placement into extraction buffer, if kept at room temperature.

Disposal

Dispose of all used test kit components and patient samples in a trash receptacle. Do not flush or pour test liquids down the drain.

Quality Control

Each INDICAID™ COVID-19 Rapid Antigen At-Home Test device has a built-in internal procedural control. The red line appearing at the “C” position is an internal procedural control. This procedural control line indicates that sufficient flow has occurred. A distinct, red-colored line should always appear if the test has been performed correctly. If the control line does not appear, the test result is invalid, and a new test should be performed using a new sample and new Test kit. If the internal procedural control line (C) is still absent after the retest, contact PHASE Scientific Technical Support at +1 (877) 934 9344 or care@indicaidusa.com.

Performing Your Test

Note

• If stored refrigerated, allow test components (test device and buffer solution vial) to equilibrate to room temperature (15–30°C or 59-86°C) before starting the Test Procedure.
• Process the collected specimen immediately after collection. Do not transport or store specimens for later testing. Inadequate specimen collection or improper handling, storage, and transport may lead to incorrect results. Do not test specimens 2 hours after collection.
• Use only the swab provided in the INDICAID™ COVID-19 Rapid Antigen At Home Test Kit.

GATHER YOUR SUPPLIES

Check the expiration date on the outside of the product box. Remove 1 swab, 1 test device pouch, and 1 buffer solution vial.
Check the buffer solution volume in the vial. If the vial is empty, DO NOT use and obtain a new buffer solution vial.

WASH YOUR HANDS
Wash your hands thoroughly for at least 20 seconds before and after handling nasal swab samples.

REMOVE ENTIRE BUFFER SOLUTION VIAL CAP
Twist off the entire cap (purple & white parts together) from the buffer solution vial. Place the vial and cap on a horizontal (flat) surface.

REMOVE NASAL SWAB FROM ITS POUCH
Remove the nasal swab from its pouch. Avoid touching the soft tip of the swab onto any surface. Only remove the swab from its pouch once the test is ready to be performed.

COLLECT NASAL SWAB SAMPLE FROM BOTH NOSTRILS USING THE SAME SWAB

Insert the entire collection tip of the swab provided (usually ½ to ¾ of an inch, or 1 to 1.5 cm) inside the nostril. Refer to diagram. Firmly sample the nasal wall by rotating the swab in a circular path against the nasal wall at least 4 times. Take approximately 15 seconds to collect the specimen. Be sure to collect any nasal drainage that may be present on the swab.

Repeat in the other nostril using the same swab
Note: With children, the maximum depth of insertion into the nostril may be less than ¾ of an inch, and you may need to have a second person hold the child’s head while swabbing.

RELEASE SAMPLE INTO BUFFER SOLUTION VIAL

Immediately place the nasal swab into the buffer solution vial.
Tilt the vial to make sure that the swab tip (soft end) is thoroughly soaked and immersed in the buffer solution.
Twist the swab back and forth 20 times in the buffer solution. Before taking out, roll the swab tip against the inner wall of the vial to release the liquid from the swab, then discard the swab.

CAP THE VIAL AND EXPOSE DROPPER TIP

Tightly cap the buffer solution vial with the vial cap.
Remove the purple part of the cap from the vial to expose the dropper tip. Avoid touching the dropper tip.

ADD BUFFER SOLUTION TO THE TEST DEVICE
Open the test device pouch and place the test device on a flat surface. Locate the sample well (S) on the test device. Slowly squeeze 3 drops of the buffer solution into the sample well.

LET TEST DEVICE SIT FOR 20 MINUTES AND READ TEST RESULTS
Start a timer for 20 minutes. Leave the test device on a table or flat surface until the timer goes off. Read the test line (T) and control line (C) results promptly at 20 minutes, and not earlier to ensure proper test performance. Results after 25 minutes should not be used.

DISPOSE OF USED TEST KIT MATERIALS
Dispose of all used test kit components and swab samples in a trash receptacle. Do not flush or pour test liquids down the drain.

Result Interpretation

• Results should be considered in the context of an individual’s recent exposures, history, and the presence of clinical signs and symptoms consistent with COVID-19.
• Test results are interpreted visually, without the aid of instruments.
• Repeat testing is needed to improve test accuracy. Please follow the table below when interpreting test results.

Status on first day of Testing| First Result Day 1| Second Result Day 3| Third Result Day 5| Interpretation
---|---|---|---|---
With Symptoms| Positive| N/A| N/A| Positive for COVID-19
Negative| Positive| N/A| Positive for COVID-19
Negative| Negative| N/A| Negative for COVID-19
Without Symptoms| Positive| N/A| N/A| Positive for COVID-19
Negative| Positive| N/A| Positive for COVID-19
Negative| Negative| Positive| Positive for COVID-19
Negative| Negative| Negative| Negative for COVID-19

POSITIVE RESULT

If the control (C) line and the test (T) line are visible, the test is positive. Any faint visible red-colored test (T) line with the control (C) line should be read as positive.

Repeat testing does not need to be performed if patients have a positive result at
any time.

A positive test result means that the virus that causes COVID-19 was detected in the sample, and it is very likely the individual has COVID-19 and is contagious. Please contact the patient’s doctor/primary care physician (if applicable) and the local health authority immediately and instruct your patient to adhere to the local guidelines regarding self-isolation. There is a very small chance that this test can give a positive  result that is incorrect (false positive).

Positive results do not rule out bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease. Individuals who test positive with the INDICAID™ COVID-19 Rapid Antigen At- Home Test should self-isolate and seek follow up care with their physician or healthcare provider as additional confirmatory testing with a molecular test for positive results may also be necessary, if there is a low likelihood of COVID-19, such as in individuals without known exposures to COVID-19 or residing in communities with low prevalence of infection.

NEGATIVE RESULT

If the control (C) line is visible, but the test (T) line is NOT visible, the test is negative.

Negative

To increase the chance that the negative result for COVID-19 is accurate, you should:

• Test again in 48 hours if the individual has symptoms on the first day of testing.
• Test 2 more times at least 48 hours apart if the individual does not have symptoms  on the first day of testing.

A negative test result indicates that the virus that causes COVID-19 was not detected in the sample. A negative result does not rule out COVID-19. There is a higher chance
of false negative results with antigen tests compared to laboratory-based tests such as PCR tests. If the test is negative but COVID-19-like symptoms, e.g., fever,
cough, and/or shortness of breath continue, follow up testing for SARS-CoV-2 with a molecular test or testing for other respiratory disease should be considered. If applicable, seek follow up care with the primary healthcare provider.

All negative results should be treated as presumptive and confirmation with a molecular assay, may be necessary if there is high likelihood of SARS-CoV-2 infection, such as in an individual with a close contact with COVID-19 or with suspected exposure to COVID-19 or in communities with high prevalence of infection. Negative results do not rule out SARS-CoV-2 infection and should not be used as the sole basis for treatment or patient management decisions, including infection control decisions.

INVALID RESULT

If the control (C) line is not visible, the test is invalid. Re-test with a new swab and new test device.

Invalid

Performance Characteristics

CLINICAL PERFORMANCE

The clinical performance of the INDICAID™ COVID-19 Rapid Antigen At-Home Test was evaluated in an on-going prospective study performed at four (4) geographically diverse sites throughout the United States. Between December 2021 and January 2022, site operators sequentially enrolled 242 eligible subjects presenting with at least one symptom of COVID-19 within 6 days of symptom onset. Using the Layuser instructions for use provided in the test kit, individuals aged 14 years and older independently collected an anterior nasal swab specimen, conducted the INDICAID™ COVID-19 Rapid Antigen At-Home Test, and interpreted and reported their self-test result. For pediatric subjects between the ages of two (2) and 13 years, an accompanying adult (e.g. parent or legal guardian aged 18 years and older) was present to collect the anterior nasal swab specimen, conduct the INDICAID™ COVID-19 Rapid Antigen At- Home Test, and interpret and report the result for the subject. A high- sensitivity FDA EUA-authorized RT-PCR SARS-CoV-2 assay was used as a comparator method to test anterior nasal swab samples that were collected from each subject by a healthcare professional.

The INDICAID™ COVID-19 Rapid Antigen At-Home Test results were compared against the results of the FDA EUA RT-PCR comparator assay to calculate the positive percent agreement (PPA) and negative percent agreement (NPA).

When conducted by a lay-user, the INDICAID™ COVID-19 Rapid Antigen At-Home Test identified 81.7% (95% CI: 71.6% – 89.4%) of the subjects that were identified as SARSCoV-2 positive by the comparator assay2 . Additionally, INDICAID™ COVID-19 Rapid Antigen At-Home Test correctly identified 99.4% (95% CI: 96.6% – 100%) of SARS CoV-2 negative subjects.

Table 1: INDICAID™ COVID-19 Rapid Antigen At-Home Test Performance Against Comparator Method (Within 6 Days Symptom Onset)

INDICAID ™

COVID-19 Rapid Antigen At-Home Test

| Comparator Method
---|---
Positive| Negative| Total
Positive| 67| 1| 68
Negative| 15| 159| 174
Total| 82| 160| 242
PPA| 81.7% (95% CI: 71.6% – 89.4%)
NPA| 99.4% (95% CI: 96.6% – 100%)

The performance of this test has not yet been clinically validated for use in patients without signs and symptoms of respiratory infection or for serial screening applications, and performance may differ in these populations.

Table 3: Positive results by age (years) of patient

Age (years)| **Total*| Comparator Positive| Prevalence| INDICAID ™ Positive**
---|---|---|---|---
2 to 13| 23| 6| 26.1%| 4
14 to 24| 34| 14| 41.2%| 14
25 to 64| 150| 54| 36.0%| 45
65+| 34| 8| 23.5%| 4

Table 4: Positive results by days since symptom onset

Days Since Symptom Onset| Cumulative Comparator Positive| Cumulative INDICAID ™ Positive| PPA
---|---|---|---
1| 13| 10| 76.9%
2| 35| 30| 82.9%
3| 52| 43| 80.8%
4| 63| 52| 81.0%
5| 74| 61| 81.1%
6| 82| 68| 81.7%

DATA ESTABLISHING PPA OF COVID-19 ANTIGEN SERIAL TESTING

A prospective clinical study was conducted between January 2021 and May 2022 as a component of the Rapid Acceleration of Diagnostics (RADx) initiative from the National Institutes of Health (NIH). A total of 7,361 individuals were enrolled via a decentralized clinical study design, with a broad geographical representation of the United States. Per inclusion criteria, all individuals were asymptomatic upon enrollment in the study and at least 14 days prior to it and did not have a SARS-CoV-2 infection in the three months prior to enrollment. Participants were assigned to one of three EUA authorized SARSCoV-2 OTC rapid antigen tests to conduct serial testing (every  48 hours) for 15 days. If an antigen test was positive, the serial-antigen testing result is considered positive.

At each rapid antigen testing time point, study subjects also collected a nasal swab for comparator testing using a home collection kit (using a 15-minute normalization window between swabs). SARS-CoV-2 infection status was determined by a composite comparator method on the day of the first antigen test, using at least two highly sensitive EUA RT-PCRs. If results of the first two molecular test were discordant a third highly sensitive EUA RT-PCR test was performed, and the final test result was based upon the majority rule.

Study participants reported symptom status throughout the study using the MyDataHelps app. Two-day serial antigen testing is defined as performing two antigen tests 36 – 48 hours apart. Three-day serial antigen testing is defined as performing three antigen tests over five days with at least 48 hours between each test.

Out of the 7,361 participants enrolled in the study, 5,609 were eligible for analysis.
Among eligible participants, 154 tested positive for SARS-CoV-2 infection based on RTPCR, of which 97 (62%) were asymptomatic on the first day of their infection, whereas 57 (39%) reported symptoms on the first day of infection. Pre-symptomatic subjects were included in the positive percent agreement (PPA) of asymptomatic individuals, if they were asymptomatic on the first day of antigen testing, regardless of whether they developed symptoms at any time after the first day of testing.

Performance of the antigen test with serial testing in individuals is described in Table 5. Table 5: Data establishing PPA of COVID-19 antigen serial testing compared to the molecular comparator single day testing throughout the course of infection with serial testing. Data is from all antigen in study combined.

Days after first PCR positive test result

| Asymptomatic on first day of testing| Symptomatic on first day of testing
---|---|---
Ag Positive / PCR Positive (Antigen Test Performance % PPA)
1 Test| 2 Test| 3 Test| 1 Test| 2 Test| 3 Test
0| 9/97| 35/89| 44/78| 34/57| 47/51| 44/47
(9.3%)| (39.3%)| (56.4%)| (59.6%)| (92.2%)| (93.6%)
2| 17/34| 23/34| 25/32| 58/62| 59/60| 43/43
(50.0%)| (67.6%)| (78.1%)| (93.5%)| (98.3%)| (100%)
4| 16/21| 15/20| 13/15| 55/58| 53/54| 39/40
(76.2%)| (75.0%)| (86.7%)| (94.8%)| (98.1%)| (97.5%)
6| 20/28| 21/27| 16/18| 27/34| 26/33| 22/27
(71.4%)| (77.8%)| (88.9%)| (79.4%)| (78.8%)| (81.5%)
8| 13/23| 13/22| 4/11| 12/17| 12/17| 7/11
(56.5%)| (59.1%)| (36.4%)| (70.6%)| (70.6%)| (63.6%)
10| 5/9| 5/8| | 4/9| 3/7|
(55.6%)| (62.5%)| (44.4%)| (42.9%)

1. Test =one(1) test performed on the noted days after first PCR positive test result. Day 0 is the first day of documented infection SARS-CoV-2.
2. Test =two(2) tests performed and average of 48 hours apart. The first test performed on the indicated day and the second test performed 48 hours later.
3. Test =three(3) tests performed and average of 48 hours apart. The first test performed on the indicated day and the second test performed 48 hours later, and a final test performed 48 hours after the second test.

LIMIT OF DETECTION (ANALYTICAL SENSITIVITY)

The limit of detection (LoD) of the INDICAID™ COVID-19 Rapid Antigen At-Home Test was determined using serial dilutions of gamma-irradiated SARS-CoV-2 virus (Isolate USA-WA1/2020, NR-52287). Contrived samples were prepared by spiking the strain into pooled human nasal matrix from presumed negative donors. 50 µl of spiked sample preparation was added onto the swab and subsequently transferred to pre-filled buffer solution vial and tested as per the IFU. The preliminary LoD initially determined by testing a two-fold dilution series of 3 replicates per concentration was confirmed by testing in 20 replicates. The confirmed LoD for the INDICAID™ COVID-19 Rapid Antigen At- Home Test 2.8 X 103 TCID50 /mL. Based upon the testing procedure for this study the LoD of 2.8 x 103 TCID50 /mL equates to 140 TCID50/swab.

The performance of this test device in the detection of the Omicron variant of SARSCoV-2 was evaluated in a dilution series of clinical specimens which were positivefor the Omicron variant. This testing was conducted by the National Institutes of Health (NIH) as a component of the Rapid Acceleration of Diagnostics (RADx®) initiative. The clinical specimens used to prepare this dilution series were not identical to the previous specimen pools prepared and tested by RADx to assess performance with the omicron variant. Results from this dilution series cannot be compared to other specimen pools and do not indicate that a test will have different clinical performance compared to other EUA authorized tests. Compared to an EUA authorized RT-PCR method, the INDICAID™ COVID-19 Rapid Antigen At-Home Test detected 100% of live virus Omicron samples at a Ct-value of 24.0 (n=5) and 40% of samples at a Ct-value of 24.8 (n=5). Testing was also compared to two additional EUAauthorized OTC antigen tests (Assay #1 and Assay #2). Omicron dilutions at lower viral concentrations (Ct-values greater than 25.8) were not detected by the INDICAID™ COVID-19 Rapid Antigen At-Home Test in this study.

Omicron Pool 2 – Live Omicron Clinical Samples

| Average N2 Ct (n=9)| Assay #1
Percent Positive (n=5)| Assay #2
Percent Positive (n=5)| INDICAID ™ COVID-19 Rapid Antigen At- Home Test Percent Positive (n=5)
---|---|---|---|---
Omicron-Dilution 1| 19.8| 100| 100| 100
Omicron-Dilution 2| 20.8| 100| 100| 100
Omicron-Dilution 3| 21.5| 100| 100| 100
Omicron-Dilution 4| 22.7| 100| 100| 100
Omicron-Dilution 5| 23.6| 100| 0| 100
Omicron-Dilution 6| 24.0| 60| 0| 100
Omicron-Dilution 7| 24.8| 0| 0| 40
Omicron-Dilution 8| 25.8| 0| 0| 0
Omicron-Dilution 9| 27.4| 0| 0| 0
Omicron-Dilution 10| 28.1| 0| 0| 0
Omicron-Dilution 11| 29.1| 0| 0| 0

CROSS-REACTIVITY (ANALYTICAL SPECIFICITY) AND MICROBIAL INTERFERENCE

Cross-reactivity and microbial interference studies were performed for related pathogens, high prevalence disease agents, and normal or pathogenic flora that are reasonably likely to be encountered in a clinical specimen for the nasal cavity. Each organism and virus (as indicated in table below) was tested in both the absence and presence of inactivated SAR-CoV2 (SARS-CoV-2 isolate USA- WA1/2020) at 3X LoD. All testing samples were prepared in pooled human nasal matrix from healthy donor. No cross reactivity or interference was observed at the concentration tested as shown in the table below.

In silico analysis was performed using the Basic Local Alignment Search Tool (BLAST) managed by the National Center for Biotechnology Information (NCBI) to estimate the likelihood of cross-reactivity with microorganisms not available for wet-testing.
The degree of protein sequence homology was determined between the SARS-CoV-2 nucleocapsid protein antigen and the following microorganisms.

• Human Coronavirus HKU1: Sequence homology between SARS-CoV-2
  nucleocapsid protein and Human Coronavirus HKU1 nucleocapsid protein is relatively low at 36.7% across 82.0% of sequences, but cross-reactivity cannot be ruled out.

• Mycobacterium tuberculosis: No protein sequence homology was found between the SARS-CoV-2 nucleocapsid protein and Mycobacterium tuberculosis total protein (5925 sequences). Homology-based cross-reactivity cannot be ruled out.

• Pneumocystis jirovecii (PJP): No protein sequence homology was found between the SARS-CoV-2 nucleocapsid protein and PJP total protein (3762 sequences). Homology-based cross-reactivity cannot be ruled out.

• SARS Coronavirus: Sequence homology between SARS-CoV-2 nucleocapsid protein and SARS-Coronavirus nucleocapsid protein was found to be 90.5% with 100% query sequence coverage. Cross-reactivity with SARS Coronavirus cannot be ruled out.

HIGH DOSE HOOK EFFECT

The INDICAID™ COVID-19 Rapid Antigen At-Home Test was tested up to 2.8 x 105 TCID50/mL (1.4 x 104 TCID50/swab) of gamma-irradiated SARS-CoV-2 (USA- WA1/2020) and no high-dose Hook Effect was observed.

ENDOGENOUS INTERFERING SUBSTANCES
The interference was performed for the potentially interfering substances that may be present in the respiratory tract or might be artificially introduced onto the nasal swab in the home environment that may cross-react or interfere with the detection of SARS-CoV-2 by the INDICAID™ COVID-19 Rapid Antigen At-Home Test. The positive (3X LoD SARS-CoV-2) and negative samples were tested with the addition of potentially interfering substances. The performance of the INDICAID™ COVID-19 Rapid Antigen At-Home Test was not affected by any of the potentially interfering substances listed in the table below at the concentration tested.

| Test Result
---|---
Potential Interferent| Test Concentration| (+) SARS-CoV-2 (3x LoD)| (-) SARS-CoV-2
Whole Blood| 4%| Positive| Negative
Mucin| 0.5%| Positive| Negative
Chloraseptic (Menthol/ Benzocaine)| 1.5 mg/mL| Positive| Negative
Naso GEL (NeilMed)| 5% v/v| Positive| Negative
CVS Nasal Drops (Phenylephrine)| 15% v/v| Positive| Negative
Afrin (Oxymetazoline)| 15% v/v| Positive| Negative
CVS Nasal Spray (Cromolyn)| 15% v/v| Positive| Negative
Zicam| 5% v/v| Positive| Negative
Homeopathic (Alkalol)| 1:10 dilution| Positive| Negative
Sore Throat Phenol Spray| 15% v/v| Positive| Negative
Tobramycin| 4 μg/mL| Positive| Negative
Mupirocin| 10 mg/mL| Positive| Negative
Fluticasone Propionate (Flonase)| 5% v/v| Positive| Negative
Tamiflu (Oseltamivir Phosphate)| 5 mg/mL| Positive| Negative
NasalCrom (Cromolyn)| 15% v/v| Positive| Negative
Nasacort (Triamcinolone)| 5 % v/v| Positive| Negative
Neo-Synephrine (Phenylephrine HCl) (Spray)| 15% v/v| Positive| Negative
Rhinocort (Budesonide)| 5% v/v| Positive| Negative
Ricola (menthol)| 1.5 mg/mL| Positive| Negative
Saline nasal spray| 15% v/v| Positive| Negative
| Test Result
---|---
Potential Interferent| Test Concentration| (+) SARS-CoV-2 (3x LoD)| (-) SARS-CoV-2
Sucrets (Dyclonine/Menthol)| 1.5 mg/mL| Positive| Negative
Zanamivir| 282 ng/mL| Positive| Negative
Zicam Cold Remedy (Galphimia glauca, Luffa)| 5% v/v| Positive| Negative
Bleach (Sodium Hypochlorite)1| 0.037% v/v| Positive| Negative
1% v/v| Negative| Negative
Dish-washing Liquid (Sodium lauryl sulfate)| 1% v/v| Positive| Negative
Hand sanitizer (ethyl alcohol)| 1% v/v| Positive| Negative
Hand Soap (Benzalkonium chloride)| 1% v/v| Positive| Negative
Laundry detergent(C12-15 pareth-7 and sodium laureth-12 sulfate)| 1% v/v| Positive| Negative
Surface Sanitizer (Citric Acid)| 1% v/v| Positive| Negative
Vicks VapoRub (Camphor, Eucalyptus oil, Menthol)| 4.7% w/w| Positive| Negative
2.6% w/w| Positive| Negative
1.2% w/w| Positive| Negative

A robust use of the INDICAID™ COVID-19 Rapid Antigen At-Home Test was
demonstrated by 9 flex studies as follows:

1. Non-level positioning of the test device
2. Varying the extraction buffer solution volume
3. Varying the swab rotation number
4. Sample volume variability
5. Result reading time variability
6. Temperature and humidity
7. Test device drop
8. Delay in sample extraction
9. Non-tilting of buffer solution vial during sample extraction

Usability Study

A usability study was conducted to evaluate the ability of representative lay users
to follow the instructional steps provided in the INDICAID™ COVID-19 Rapid Antigen

At-Home Test Quick Reference Guide (QRG) under expected use conditions, comprehend the potential set of test results, and understand the product labeling. Thirty (30) representative test kit users (self-testers, caregiver- child, and proxy caregiver-adult pairs) were observed performing an INDICAID™ COVID-19 Rapid Antigen At-Home Test while following QRG, and other instructional materials, that accompany the kit. Participants were asked to collect a nasal swab sample to perform the test. They were also asked to interpret a mock test result and state the appropriate course of action based upon the test result. Participants additionally were provided with a written questionnaire to evaluate their understanding of the product labeling and to provide subjective feedback about the ease-of-use and perceived safety of the kit.

Successful completion of each study task performed by the subjects was determined by unassisted professional observation. Participants correctly performed 96.7% (667/690) of the steps/tasks and 96.7% (29/30) of the participants correctly interpreted the test results. 93.1% (27/29) of the participants provided a fully correct follow-up action response given their test result, while the remaining 6.9% (2/29) provided a partially correct response. For label comprehension questions, 94.4% (170/180) were answered correctly across all subjects.

Technical Support

For more information, questions, or support, please visit www.indicaidusa.com,
or contact us at:

• Telephone: +1 (877) 934 9344
• Email: care@indicaidusa.com

Ordering and Contact Information

PHASE Scientific International, Ltd.

• Tel: 877-934-9344
• Email: care@indicaidusa.com

Symbols

For prescription use only
In vitro diagnostic medical device
Consult Instructions for use
Caution—consult accompanying documents
Temperature limitation
Keep away from sunlight
Sufficient for use
Keep away from moisture
Do not reuse
 Catalog number
 Batch code
Use by
 Manufacturer

References

• Coronavirus Disease 2019 (COVID-19) | CDC
• INDICAID® COVID-19 Rapid Antigen Tests (OTC)｜INDICAID
• U.S. Food and Drug Administration
• 800-222-1222" class="nturl" > Poison Control | Call Poison Help Centers Now 1-800-222-1222

Read User Manual Online (PDF format)

Download This Manual (PDF format)

Download this manual  >>