LUCIRA NAAT Nucleic Acid Amplification Test Instruction Manual

: June 9, 2024
: Lucira

Table of Contents

NAAT Nucleic Acid Amplification Test
References
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NAAT Nucleic Acid Amplification Test

COVID-19 & Flu Home Test
Nucleic Acid Amplification Test (NAAT)
For Individuals with signs and symptoms of Respiratory Tract Infection including COVID-19

Table of Contents

Intended Use

3

Summary and Explanation of the Test

4

Principles of the Procedures

4

Warnings and Precautions

5

Section A – Reagents and Materials

6

Section B – Directions for running the Lucira COVID-19 & Flu Home Test

7

Section C – Test Unit Result Display

10

Section D – Quality Control Testing for Point of Care Settings

14

Section E – Quality Systems Evaluation

15

Performance Characteristics

16

Limitations

42

Technical Assistance

43

References

43

Table of Symbols

43

Instructions For Use (IFU) INST095 Rev. 6

2

LuciraTM COVID-19 & Flu Home Test
Instructions for Use
For Use under Emergency Use Authorization (EUA) only For in vitro Diagnostic Use For Use with Self-collected Nasal Swab Specimens in individuals aged 14 years or older For Use with Adult-collected Nasal Swab Specimens in individuals aged 2 years or older For Over-the-Counter (OTC) Use
Intended Use
The Lucira COVID-19 & Flu Home Test is a single use (disposable) home test kit intended for simultaneous rapid in vitro qualitative detection and differentiation of SARS-CoV-2, influenza A, and influenza B viral nucleic acid. This test is authorized for non-prescription home use with anterior nasal swab samples from individuals 14 years or older (self-collected) or individuals 2 years or older (collected by an adult) with signs and symptoms consistent with a respiratory tract infection, including COVID-19. Clinical signs and symptoms of respiratory viral infection due to SARS-CoV-2 and influenza can be similar.
The Lucira COVID-19 & Flu Home Test is intended for use in the differential diagnosis of SARS-CoV-2, influenza A, and influenza B in clinical specimens and is not intended to detect influenza C. SARS-CoV-2, influenza A, and influenza B viral nucleic acid is generally detectable in anterior nasal swab samples during the acute phase of infection.
Positive results indicate the presence of viral nucleic acid, 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 pathogens not detected by the test. The agent detected may not be the definitive cause of disease. Individuals who test positive with the Lucira COVID-19 & Flu Home Test should self-isolate and seek follow up care with their physician or healthcare provider as additional testing may be necessary.
Negative results for SARS-CoV-2 and influenza B are presumptive, meaning that they should be confirmed, if necessary for patient management, with an authorized or cleared molecular test performed in a CLIA-certified laboratory that meets requirements to perform high or moderate complexity tests.
Negative results do not rule out SARS-CoV-2, influenza A, and/or influenza B infection and should not be used as the sole basis for treatment or other management decisions, including infection control decisions. Negative results should be considered in the context of current prevalence of infection, an individual’s recent exposures, history and the presence of clinical signs and symptoms consistent with respiratory infection. Individuals who test negative and continue to experience symptoms of fever, cough and/or shortness of breath may still have a respiratory infection and should seek follow up care with their healthcare provider.
Individuals should report 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 SARS-CoV-2 Tests provided by CDC.
The Lucira COVID-19 & Flu Home Test is authorized for non-prescription self- use by individuals aged 14 years or older and/or, as applicable, for an adult lay user testing another person aged 2 years or older in a non-laboratory setting. The Lucira COVID-19 & Flu Home Test is only for use under the Food and Drug Administration’s Emergency Use Authorization.

Instructions For Use (IFU) INST095 Rev. 6

3

Summary and Explanation of the Test
The Lucira COVID-19 & Flu Home Test is a rapid, instrument-free, single-use molecular diagnostic test for the qualitative detection of SARS-CoV-2, Influenza A, and Influenza B RNA from nasal swab samples in individuals with known or suspected COVID-19 or flu. The test contains all the components required to perform testing.
Principles of the Procedures
The Lucira COVID-19 & Flu Home Test utilizes RT-LAMP technology to detect RNA of SARS-CoV-2, Influenza A, and Influenza B. This technology can create a signal from a few copies of RNA in less than 30 minutes. The RT-LAMP amplification reaction occurs in two phases, a non-cyclic phase followed by a cyclic phase. During the non-cyclic phase, reverse transcriptase, with RNase H activity, converts the RNA target into cDNA. A DNA polymerase with strand displacement activity then amplifies the cDNA. A successful amplification reaction creates a pH change and subsequently a color change of the halochromic agents within the reaction mixture.
The Sample Vial contains an elution buffer that allows the swab contents to be eluted and lysed at room temperature, releasing viral and human RNA for downstream detection. Upon engagement of the Sample Vial and Test Unit, this eluant enters a fluidic module contained within the Test Unit that has several individual reaction chambers. The eluant resolubilizes lyophilized reagents contained within these chambers, which are needed to perform the RT-LAMP reaction. An internal electronic heating element detects this chamber filling and automatically turns on, initiating amplification within the reaction chambers. The reactions are confined within the fluidic unit and no other part of the Test Unit has contact with the sample during amplification.
The Test Unit contains two chambers that target SARS-CoV-2 RNA, two chambers that target Flu A, two chambers that target Flu B, and one chamber for a control (TIC). For SARS-CoV-2, the test targets two non-overlapping regions in the N gene and Orf7b/8 gene. For Influenza A, the test targets one region of Segment 5, two non-overlapping regions of Segment 7, and one region of Segment 8. For Influenza B, the test targets one region of Segment 5 and one region of Segment 8. For Influenza B, the test targets one region of Segment 5 and one region of Segment 8.
The color change of the reaction mixture is detected in real time using optical and electronic elements contained within the Test Unit. An on-board microprocessor analyzes the color change data to detect the presence of amplification, and hence the target RNA, in each chamber. A diagnostic algorithm, included in the device firmware, is then used to determine patient infectivity status and the results are shown via LED indicators. Results for the test are displayed as either positive, negative, or invalid. A positive result may show in as few as 11 minutes; a negative or invalid result will display in 30 minutes. The result display persists for a minimum of 8 hours and a maximum of 12 hours after the test has finished running.

Instructions For Use (IFU) INST095 Rev. 6

4

WARNINGS AND PRECAUTIONS
· For in vitro diagnostic use · For use under FDA Emergency Use Authorization (EUA) only. · This product has not been FDA cleared or approved, but has been authorized for emergency use by FDA
under an EUA · This product has been authorized only for the detection and differentiation of nucleic acid from
SARS-CoV-2, Influenza A, and Influenza B, not for any other viruses or pathogens; and, · 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 Federal Food, Drug and Cosmetic Act, 21 U.S.C. § 360bbb-3(b)(1), unless the declaration is terminated or authorization is revoked sooner.
· Leave test components sealed in foil pouch until just before use. · Proper sample collection and sample handling are essential for correct results. · Do not touch swab tip when handling swab sample. · Do not use any components with visible damage. · Use the product only as specified, without modification, or the protection supplied by the product can be
compromised. · Do not use components after their expiration date. · Choose a level location to do this test where you can let the test sit undisturbed (out of reach of pets,
pests, or children) for 30 minutes. · The device may be hot to touch after the test is done. · Do not place the test closer than 30cm (12 inches) to devices or appliances, such as antenna cables and
external antennas, that may cause interference while the test is running.
· All kit components are single use items. Do not use with multiple specimens. · Do not try to disassemble the Test Unit. · The elution buffer may contain irritants. Do not ingest the contents of the tube. If the contents of the tube
are splashed in your eyes, flush your eyes with water. If the contents splash onto your skin, wash with soap and water. If irritation persists, notify a health care provider.
· After use remove the batteries, place the test unit in plastic disposal bag and dispose all test kit materials in trash. Do not allow the test unit to come into contact or be disposed of with bleach, as harmful gases could be emitted as a result.
· At low frequency, clinical samples contain inhibitors that may generate invalid results. · Performance characteristics of this test have been established with specimen types listed in the Intended
Use section only. The performance with other specimen types or samples has not been validated. · Only use the components provided. Do not use swabs from the other tests.

Instructions For Use (IFU) INST095 Rev. 6

5

SECTION A – Reagents and Materials
LuciraTM COVID-19 & Flu Home Test contents:
· Package Insert · Nasal Swab: one sterile flocked nasal swab in a peel-pouch; · Sample Vial: a single-use, disposable vial containing an elution buffer to release and lyse virions
from a nasal swab sample; · Test Unit: a single-use, disposable unit with lyophilized reagents for multiplexed amplification
and electronic readout for detection of SARS-CoV-2, Flu A, and Flu B RNA; · Batteries: two AA batteries for the Test Unit; and · Plastic disposal bag to dispose of the test after use.
NOTE: For optimal performance, use the swabs provided in the test. Other swabs are not suitable for use with this test.
STORAGE AND HANDLING
· Tests must always be stored at temperature between 15-30°C / 59-86°F. · Tests must be stored at ambient humidity 10%-80%. · IP21: The Test Unit has an enclosure protection rating of IP21. This means the Test Unit has
protection from the insertion of a finger or solid objects greater than 1/2″ (12.5mm) in diameter. This also means the Test Unit has protection against vertically falling drops of water or condensation. · Do not reuse test components. · Do not remove the Test Unit from the foil pouch until immediately before use.

Instructions For Use (IFU) INST095 Rev. 6

6

Section B Directions for running the Lucira COVID-19 & Flu Home Test

· Choose a location to do this test where it can sit UNDISTURBED for 30 minutes.
· Please read all instructions carefully before you begin. · Do not insert batteries into test unit until ready to
perform test. · Keep test box to create a personal verified digital record
of your test result. · Make sure your test kit contains: 2 AA
batteries, test unit (pouch 1), sample vial (pouch 2), swab (labeled 3), and plastic disposal bag. · Wash and dry hands.
1. Set Up Test
· When ready to begin test, open test unit pouch 1.
Open battery door and insert batteries. Check that Ready light is on.
· Open sample vial pouch 2.
REMOVE sample vial seal
then GENTLY set in test unit but do NOT push the vial down.

2. Swab Both Nostrils
For this test to work properly, it is important to swab BOTH nostrils.
· Remove swab and hold with handle end. Do not set swab
down.
· Tilt head back and gently insert swab tip until it is fully
inside your nostril and you meet resistance. o For adults – insert less than one inch. o For younger children – insert less than 1/2 inch
· Once swab tip is fully inside nostril, roll the swab
5 times around the inside walls of your nostril.
The swab should be touching the walls of the nostril as you rotate.
· Repeat swab step in other nostril.
5 Rotate x in BOTH nostrils.
Make sure to roll around inside walls to collect a good sample.

Ready

COVID

Neg Flu A Pos

–

Flu B

Note: Keep vial away from children. Avoid contact with eyes and skin. If contact occurs, rinse with water. If irritation persists, seek medical attention.
Instructions For Use (IFU) INST095 Rev. 6

Adults must swab children ages 2 years or older.
7

3. Stir Swab and Run Test

15x

· Insert swab into the sample vial until it touches the bottom.
· Mix sample by stirring around the sample vial 15 times.
·Discard swab.

Ready

COVID

Neg Flu A Pos

–

Flu B

· Immediately snap cap closed and press vial down into test unit until it clicks.
· Ready light will start blinking when test is running.

CLICK

Ready

COVID

Neg Flu A Pos

–

Flu B

READY

If Ready light is not blinking within 5 seconds, use palm of your hand to press down more firmly to start test.
Do not move test unit once the test has started running.
Wait 30 minutes.

Instructions For Use (IFU) INST095 Rev. 6

4. Read Result
· Ready light will continue blinking while the test is running.
· Positive results may display before the test is done running; however, you must wait until the Ready light has stopped blinking to interpret all test results.
· Results may be positive for more than one virus. · Ready light will turn off and all results for COVID-19, Flu
A, and Flu B will display in 30 minutes when test is done.

Example Result: Positive for COVID-19 & Flu A; Negative for Flu B.
See Section C for all possible results.

Ready

COVID

Neg Flu A Pos

–

Flu B

POSITIVE Results

Positive results light up on the right

COVID-19 Positive

Flu A Positive

Flu B Positive

Ready

COVID

Neg Flu A Pos Neg Flu A Pos Neg Flu A Pos

–

Flu B

–

Flu B

–

Flu B

NEGATIVE Results

Negative results light up on the left

COVID-19 Negative Flu A Negative

Flu B Negative

Ready

COVID

Neg Flu A Pos Neg Flu A Pos Neg Flu A Pos

–

Flu B

–

Flu B

–

Flu B

INVALID Results

Positive and Negative lights flash/blink if result is Invalid

Ready

Invalid results may occur for one, two or all three

COVID

viruses. If an invalid result is observed for any of the

Neg Flu A Pos viruses the entire test is considered invalid. Retest

–

Flu B + with a new test and use the retest results as final.

If you receive any invalid results, retest or contact Lucira at 1-888LUCIRA-4 to obtain a free replacement test.

8

Lucira Connect is a website that helps you:
· Create a shareable digital record of your test result · Connect to telehealth from home to discuss care and treatment options with a healthcare
provider Use your smartphone camera to scan the QR code on the top of the test unit or on the
sticker on the back of the box to begin at luciraconnect.com

If the test is POSITIVE
It is very likely you have COVID-19 (if the test result is positive for COVID-19) or flu (if the test result is positive for flu A or flu B) and it is important to be under the care of a healthcare provider. It is likely you will be asked to isolate yourself at home to avoid spreading the virus to others. There is a very small chance that this test can give a positive result that is wrong (a false positive). Your healthcare provider will work with you to determine how best to care for you based on your test results along with medical history and your symptoms.
If the test is NEGATIVE
A negative result means the virus that causes COVID-19 (if you test negative for COVID-19) or flu (if you test negative for flu A & flu B) was not found in your sample. However, it is possible for this test to give a negative result that is incorrect (a false negative) in some people with COVID-19 or flu. This means you could possibly still have COVID-19 or flu even though the test is negative. If this is the case, your healthcare provider will consider the test result with all other aspects of your history such as symptoms and possible exposures to decide how to care for you. It is important you work with your healthcare provider to help you understand the next steps you should take.
If the test is INVALID An invalid result means something with the test did not work properly. If the test result is invalid, the positive and negative lights on the device will be blinking/flashing. If your test shows an invalid result, retest or contact Lucira at 1-888- LUCIRA-4 to obtain a free replacement test.

5. Dispose of Test
After use remove the batteries, place the test unit in plastic disposal bag and dispose all test kit materials in trash. Do not allow the test unit to come into contact or be disposed of with bleach, as harmful gases could be emitted as a result.

Instructions For Use (IFU) INST095 Rev. 6

9

Section C – Test Unit Result Display

Test is ready to be run

Test is running

Blinking/flashing

COVID-19 Flu A Flu B

Negative Negative Negative

Positive Positive Positive

COVID-19 Flu A Flu B

Positive Negative Negative

Negative Positive Negative

Instructions For Use (IFU) INST095 Rev. 6

Negative Negative Positive
10

COVID-19 Flu A Flu B

Positive Positive Negative

Positive Negative Positive

Negative Positive Positive

COVID-19 Flu A Flu B

Positive Not available yet Not available yet

Not available yet Positive Not available yet

Not available yet Not available yet Positive

COVID-19 Flu A Flu B

Positive Not available yet Positive

Not available yet Positive Positive

Instructions For Use (IFU) INST095 Rev. 6

Positive Positive Not available yet
11

Covid-19, Flu A, Flu B — Invalid Results
If any individual assay gives an invalid result, all results for that Test Unit should be considered invalid and testing should be repeated with a new Lucira COVID-19 & Flu Home Test.

Invalid

Instructions For Use (IFU) INST095 Rev. 6

Invalid
12

Covid-19, Flu A, Flu B — Invalid Results
If any individual assay gives an invalid result, all results for that Test Unit should be considered invalid and testing should be repeated with a new Lucira COVID-19 & Flu Home Test.

Invalid

Instructions For Use (IFU) INST095 Rev. 6

Invalid
13

Section D – Quality Control Testing for Point of Care Settings
External run controls (ERCs) are not required to use this test kit.

Instructions For Use (IFU) INST095 Rev. 6

14

Section E – Quality Systems Evaluation
The Quality Systems of Lucira Health, Inc. were independently evaluated. The evaluation has provided evidence to establish that the quality systems and manufacturing capability are likely to achieve the performance noted in this labeling.

Instructions For Use (IFU) INST095 Rev. 6

15

PERFORMANCE CHARACTERISTICS

Limit of Detection (LoD) – Analytical Sensitivity
The limit of detection was determined for 5 human derived viral isolates individually (referred to as anchor strains): 1. Influenza A H3N2: A/HongKong/4801/2014 2. Influenza A H1N1pdm09: A/Michigan/45/2015 3. Influenza B, Yamagata Lineage: B/Phuket/3073/2013 4. Influenza B, Victoria Lineage: B/Colorado/6/2017 5. SARS-CoV-2: Heat Inactivated 2019-nCoV/USA-WA1/2020

Each virus was serially diluted into Natural Nasal Swab Matrix (NNSM), pipetted onto a fresh, unused nasal swab, and run on two device lots. NNSM was prepared by pooling negative patient specimens in viral transport media, previously tested negative for SARS-CoV-2, Influenza A, and Influenza B. The preliminary LoD for the device was determined by testing at least three (3) target concentrations at 2-fold dilutions on each lot of devices. For each lot, each concentration was tested in replicates of seven (7) devices by three (3) unique operators, for a total of 21 replicates per concentration. Additionally, each operator ran two (2) Non-Template Controls (NTC) as negative controls immediately after each target concentration. The LoD for each lot was separately determined as the lowest concentration that yielded greater than 95% positive results. At least one of the concentrations run had to produce < 95% positive results for the virus. The preliminary LoD for the device was defined as the higher LoD of the two lots.

The LoD was confirmed by testing 20 replicates at the preliminary LoD concentration on a single lot for each target. Two (2) additional operators, who were not involved in determining the preliminary LoD, performed the confirmation testing by each running ten (10) devices from one lot at the determined preliminary LoD concentration. Each virus produced 19/20 positive replicates, confirming the LoD for each virus. Detailed results are shown in Table 1 through 5 and summarized in Table 6 below.

Table 1. LoD Determination Results SARS-CoV-2

Genome equivalents /
mL VTM*

Genome equivalents /
swab

SARS-CoV-2: Heat Inactivated 2019-nCoV/USA-WA1/2020

Positive/Total Valid

Preliminary Lot 1

Preliminary Lot 2

Confirmatory

Percent Positive

Preliminary Lot 1

Preliminary Lot 2

Confirmatory

727

2180

21/21

—

100%

—

363

1090

21/21

20/21

20/20

100%

95%

100%

181

544

21/21

19/21

—

100%

90%

—

91

272

15/21

14/21

—

71%

67%

—

Since most tests utilize viral transfer media (VTM) as a matrix to elute the swab, the concentrations of genome equivalents per swab were also converted to corresponding concentrations of genome equivalents per mL of VTM (assuming 100% elution of a swab into 3 mL of VTM). For example, the concentration of 1260 genome equivalents (GE) per swab corresponds to 420 copies per mL of VTM.

Instructions For Use (IFU) INST095 Rev. 6

16

Table 2. LoD Determination Results Influenza A H1N1pdm09

Genome equivalents /mL VTM*

Genome equivalents
/swab

Influenza A H1N1pdm09: A/Michigan/45/2015

2500

7500

1250

3750

627

1880

313

938

Positive/Total Valid

Percent Positive

Preliminary Lot 1 21/21 21/21 19/21 13/21

Preliminary Lot 2 21/21 21/21 19/21 15/21

Confirmatory
-20/20
—

Preliminary Lot 1
100.00% 100.00%
90% 62%

Preliminary Lot 2
100.00% 100.00%
90% 71%

Confirmatory
-100%
—

Table 3. LoD Determination Results Influenza A H3N2

Genome equivalents /mL VTM*

Genome equivalents /swab

Influenza A H3N2 A/HongKong/4801/2014

1680

5040

840

2520

420

1260

210

630

Positive/Total Valid

Percent Positive

Preliminary Lot 1
21/21 21/21 21/21 18/21

Preliminary Lot 2
21/21 21/21 20/21 18/21

Confirmatory
–19/20 —

Preliminary Lot 1
100% 100% 100% 86%

Preliminary Lot 2
100% 100% 95% 86%

Confirmatory
–95% —

Table 4. LoD Determination Results Influenza B, Victoria Lineage

Genome equivalents /mL VTM*

Genome equivalents /swab

Influenza B, Victoria Lineage: B/Colorado/6/2017

5767

17300

2890

8670

1443

4330

723

2170

Positive/Total Valid

Percent Positive

Preliminary Lot 1 21/21 21/21 20/21 13/21

Preliminary Lot 2 21/21 21/21 21/21 20/21

Confirmatory
–20/20 —

Preliminary Lot 1 100% 100% 95% 62%

Preliminary Lot 2 100% 100% 100% 95%

Confirmatory
–100% —

Instructions For Use (IFU) INST095 Rev. 6

17

Table 5. LoD Determination Results Influenza B, Yamagata Lineage

Genome equivalents /mL VTM*

Genome equivalents
/swab

Influenza B, Yamagata Lineage: B/Phuket/3073/2013

1690

5070

847

2540

423

1270

211

634

Positive/Total Valid

Percent Positive

Preliminary Lot 1 20/21 14/21 15/21 13/21

Preliminary Lot 2 21/21 20/21 18/21 16/21

Confirmatory
20/20 —-

Preliminary Lot 1 95% 67% 71% 62%

Preliminary Lot 2 100% 95% 86% 76%

Confirmatory
100% —-

Assay and Subtype or Lineage
COVID-19 Flu A, H1N1pdm09 Flu A, H3N2 Flu B, Victoria Lineage Flu B, Yamagata Lineage

Table 6. LoD Summary

Target

Limit of Detection (GE/swab)

2019-nCoV/USA-WA1/2020 A/Michigan/45/2015 A/Hong Kong/4801/2014 B/Colorado/6/2017 B/Phuket/3073/2013

1090 3750 1260 4330 5070

Limit of Detection (GE / mL VTM equivalents)*
363 1250 4200 1440 1690

The LoD for SARS-CoV-2, Influenza A H3N2: A/HongKong/4801/2014, Influenza A H1N1pdm09: A/ Michigan/45/2015, Influenza B, Yamagata Lineage: B/Phuket/3073/2013 and Influenza B, Victoria Lineage: B/ Colorado/6/2017 were determined to be 1090, 1260, 3750, 5070 and 4330 Genome equivalent/swab (GE/swab), respectively. Additional studies confirmed the LoD as determined by preliminary results.

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Co-spike LoD Equivalency Study
To demonstrate that a co-spike of 3 viral targets does not impact the limit of detection, a confirmatory LoD study at the established LoD of 1X LoD was performed. All three targets (Co-spike 1: Influenza A/Michigan/45/2015, Influenza B/ Colorado/6/2017 and SARS-CoV-2 and Co-spike 2: Influenza A/Hong Kong/4801/2014, Influenza B/ Phuket/3073/2013 and SARS-CoV-2) were diluted in NNSM to a concentration of 3X LoD and then pooled together to form a co-spike at 1X LoD in NNSM. The NNSM containing all three targets at 1X LoD was then pipetted onto a fresh, unused nasal swab and tested per the instructions for use. The results demonstrated that 95% (for each of the three targets) of replicates were positive at 1X LoD, indicating that the LoD is confirmed in a triple co-spike and a cospike of all three targets is acceptable to use for additional studies. The presence of all three target analytes in a specimen did not adversely affect the analytical sensitivity of the Lucira COVID-19 & Flu Home Test.

Cospike
Co-spike 1 Co-spike 2

Target 1
A/Michigan/45/2015 A/Hong Kong/4801/2014

Table 7. Co-spike LoD Confirmation

Target 2 B/Colorado/6/2017

Target 3 SARS-CoV-2

Flu A POS/ Total Valid
38/40

B/Phuket/3073/2013 SARS-CoV-2

20/20

Flu B POS/
39/40 20/20

COVID-19 POS / Total Valid
40/40
20/20

Inclusivity (Analytical Reactivity)
a) Wet Testing The inclusivity of the Lucira assays was evaluated with 20 Influenza A strains (10 H1N1 pdm09 and 10 H3N2), 10 Influenza B strains (5 Yamagata and 5 Victoria lineages), and 3 SARS-CoV-2 strains representing temporal, geographic, and genetic diversity within the currently circulating subtypes and lineages. At least 2 strains from the last 5 years were selected for Influenza A H1N1pmo09, Influenza A H3N2 and Influenza B. All Influenza strains were quantified by an in-house, validated qPCR assay to standardized concentration units. SARS-CoV-2 strains were quantified by ddPCR by the supplier. All strains were individually tested at 3X LoD in 3 replicates to demonstrate inclusivity. The results are shown in Tables 8 through 10 below.

Table 8. COVID-19 Assay Results with Tested SARS-CoV-2 Strains

Target

SARS-CoV-2 isolate 5574/2020

Alpha Variant

SARS-CoV-2 isolate 015421/2021

Beta Variant

SARS-CoV-2 isolate hCoV-19/USA/MD-HP05285/2021 Delta Variant

SARS-CoV-2 strain 2019-nCoV/USA-WA1/2020

Anchor CoV

Test Concentration (cp/swab) 3275
3275 3275 3275

COVID-19 POS / Total Valid 3 / 3
3 / 3 3 / 3 3 / 3

% Positive
100% 100% 100% 100%

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Table 9. Flu A Assay Results with Tested Influenza A Strains

Target
A/Indiana/02/2020 A/Hawaii/66/2019 A/Victoria/2570/2019 A/Wisconsin/588/2019 A/Michigan/45/2015 A/Bangladesh/3002/2015 A/Dominican/Republic/7293/2013 A/Iowa/53/2015 A/Christchurch/16/2010 A/California/7/2009 A/New York/21/2020 A/Tasmania/503/2020 A/Hong Kong/2671/2019 A/Hong Kong/45/2019 A/Singapore/INFIMH-16-0019/2016 A/Hong Kong/4801/2014 A/Switzerland/9715293/2013 A/Brisbane/10/2007 A/Texas/50/2012 A/Perth/16/2009

Subtype
H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09 H1N1pdm09
H3N2 H3N2 H3N2 H3N2 H3N2 H3N2 H3N2 H3N2 H3N2 H3N2

Test Concentration (cp/swab)
11250

Flu A POS / Total Valid 3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

11250

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

3785

3 / 3

%Positive
100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100% 100%

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Table 10. Flu B Assay Results with Tested Influenza B Strains

Target
B/Washington/02/2019 B/Colorado/6/2017 B/Florida/78/2015 B/Texas/02/2013 B/Michigan/09/2011 B/Texas/81/2016 B/Phuket/3073/2013 B/Montana/05/2012 B/Massachusetts/02/2012 B/Wisconsin/1/2010

Lineage
Victoria Victoria Victoria Victoria Victoria Yamagata Yamagata Yamagata Yamagata Yamagata

Test Concentration Flu B POS / Total Valid (cp/swab)

13000

3 / 3

13000

3 / 3

13000

3 / 3

13000

3 / 3

13000

3 / 3

15200

3 / 3

15200

3 / 3

15200

3 / 3

15200

3 / 3

15200

3 / 3

%Positive
100% 100% 100% 100% 100% 100% 100% 100% 100% 100%

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b) In silico
i) SARS-CoV-2 Predicted Reactivity Inclusivity of the Lucira SARS-CoV-2 Assay was demonstrated by in-silico reactivity of the assay against publicly available SARS-CoV-2 strains using the assay’s primers. SARS-CoV-2 sequences were downloaded from the Global Initiative on Sharing All Influenza Data (GISAID, https:// www.gisaid.org) database monthly from December 1, 2020 through January 15, 2023. As of April 15, 2021, all SARS-Cov-2 sequences uploaded to GISAID each month are downloaded and up to 50,000 sequences are sampled. Prior to the April 15, 2021 datapoint, all sequences uploaded that month were included in the analysis. For each sample, sequences were imported into Geneious and trimmed to remove ambiguous bases, filtering by length post- trim to ensure coverage of target regions. Geneious was then used to predict primer binding, and binding results were analyzed to apply reactivity rules. Reactivity for a set was defined as having at most one mismatch on a primer, and no mismatches within 5 nucleotides of the leading edge for each primer. A single nucleotide mismatch in one of the primers for LAMP assays is not expected to impact the limit of detection, unless it is in the leading end of the primer as previously demonstrated by work on MERS-CoV (PMID 25103205). Between December 1, 2020 and January 15, 2023, 1,068,203 sequences were analyzed and 99.97% were found to be reactive.

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Table 11. SARS-CoV-2 Reactivity Results by Month

Sequence Dates Dec 1 Dec 31, 2020
Jan 1 Jan 31, 2021 Feb 1 Feb 28, 2021 Mar 1 March 31, 2021 Mar 15 Apr 15, 2021 Apr 15 May 15, 2021 May 15 Jun 15, 2021 Jun 15 Jul 15, 2021 Jul 15 Aug 15, 2021 Aug 15 Sep 15, 2021

N 34,775
25,824 42,888 72,943 49,611 48,435 49,365 48,252 48,650 48,547

No. Passing Sequences 34,761
25,808 42,885 72,936 49,597 48,424 49,341 48,241 48,646 48,545

Sep 15 Oct 15, 2021

49,886

49,870

Oct 15 Nov 15, 2021

48,804

48,797

Nov 15 Dec 15, 2021

47,592

47,583

Dec 15, 2021 Jan 15, 2022

48,105

48,099

Jan 15 Feb 15, 2022

47,011

46,999

Feb 15 Mar 15, 2022

45,907

45,900

Mar 15 Apr 15, 2022 Apr 15 May 15, 2022

46,605 45,870

46,595 45,844

May 15 Jun 15, 2022

44,542

44,497

Jun 15 Jul 15, 2022

46,113

46,099

Jul 15 Aug 15, 2022

23,857

23,845

Aug 15 Sep 15, 2022

25,003

24,997

Sep 15 Oct 15, 2022

11,306

11,301

Oct 15, 2022 – Dec 15, 2022 Dec 15, 2022 Jan 15, 2023

41,491 26,821

41,478 26,814

All time points (Dec 1, 2020 to Jan 15, 2023)

1,068,203

1,067,902

Percent Passing 99.96% 99.94% 99.99% 99.99% 99.97% 99.98% 99.95% 99.98% 99.99% 100.00% 99.97% 99.99% 99.98% 99.99% 99.97% 99.98% 99.98% 99.94% 99.90% 99.97% 99.95% 99.98% 99.96% 99.97% 99.97%
99.97%

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ii) SARS-CoV-2 Reactivity Results by Variant For SARS-CoV-2, reactivity was also predicted for each Greek-letter variant monitored by WHO or CDC. Up to 10,000 sequences are sampled each month for each Greek-letter variant. Reactivity predictions based on established rules were tabulated by variant for the sample of SARS-CoV-2 sequences uploaded to GISAID between December 15, 2022, and January 15, 2023. The Lucira COVID-19 & Flu Home Test is predicted to be reactive to all variants identified. Specifically, within this dataset, 8776/8779 (99.97%) Omicron sequences were predicted to be reactive to the assay demonstrating that the Lucira COVID-19 & Flu Home test is reactive to the Omicron variant.

Variant
Alpha Beta Delta Gamma Omicron Epsilon Eta Kappa Mu Zeta Total

Table 12. SARS-CoV-2 Reactivity Results by Variant

No. Sequences Downloaded (post-filter)
1854 44 5800 1098
8779 51 18 11 1 238 17894

No. Sequences Reactive In-Silico (Percent)
1854 ( 100.00%) 44 (100.00%) 5799 (99.98%) 1098 (100.00%)
8776 (99.997%) 51( 100.00%) 18( 100.00%) 11( 100.00%) 1( 100.00%) 238 ( 100.00%) 17,890 (99.998%)

iii) Influenza Predicted Reactivity Sequences were downloaded for each targeted segment for each subtype: A/H3N2, A/pH1N1, B/Victoria, and B/ Yamagata. Sequences were imported to Geneious and trimmed to remove ambiguities, and then filtered by length to include only whole segment sequences. Primer binding with both primer sets was predicted using Geneious and results were analyzed using R to apply reactivity rules. Reactivity was defined as having at least one primer set with at most one SNP on each primer, and no SNPs within 5 nucleotides of the leading edge for each primer. All four subtypes had over 95% of sequences reactive for both the last year (December 2021 January 2023) and the last 3 years (December 2019 January 2023). All are also reactive to over 95% of sequences in the last 5 years (2017-2023).

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Years
Dec 2016-Dec 2017 Dec 2017-Dec 2018 Dec 2018-Dec 2019 Dec 2019-Dec 2020
Dec 2020-Dec 2021 Dec 2021-Jan 2023 Dec 2019-Jan 2023
[Last 3 years] Dec 2017 – Jan 2023
[Last 5 years]

Table 13. Predicted Reactivity per Influenza Target

A/H3N2

A/pH1N1

B/Victoria

Reactive

N

Reactive

N

Reactive

N

99.5%

5,754

98.7%

1,123

99.5%

1,447

99.1%

4,097

96.8%

3,456

89.1%*

829

98.4%

5,268

98.3%

4,746

93.9%

2,409

97.4%

1,157

99.7%

2,976

97.8%

3,627

98.1% 97.7%

270 15,062

98.5% 95.5%

194 2,271

99.2% 99.6%

1,007 531

97.7%

16,489

98.0%

5,441

98.3%

5,165

98.1%

25,854

97.8%

13,643

96.1%

8,403

B/Yamagata

Reactive

N

99.6%

1562

99.8%

3335

99.7%

675

98.2%

112

N/A

98.2%

112

99.7%

4,122

Years: Defined as Dec 1st of start year to Nov 30th of the end year. 2022 data covers through December 15, 2022.

*Wet testing confirmed assay performance with B/Colorado/6/2017 as reported in Table 10.

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Cross-Reactivity (Analytical Specificity)

a) Wet Testing

The specificity of the assay was evaluated in cross-reactivity testing using 29 potential pathogens or

commensal organisms. For each organism, 35 L of undiluted organism was spiked onto a nasal swab with

35 L of NNSM. The swab was then eluted and run on the Lucira test. As shown below, the cross-reactivity

testing confirmed that none of the organisms were cross reactive with the Lucira COVID-19 & Flu Home Test at

the concentrations tested.

Table 14. Cross-Reactivity Results

Microbial Target

Test Concentration

COVID-19 (# POS / #
Tested)

Parainfluenza virus 1

1.26E+06 TCID50/mL

0/3

Parainfluenza virus 2

1.60E+06 TCID50/mL

0/3

Parainfluenza virus 3

8.51E+07 TCID50/mL

0/3

Parainfluenza virus 4

1.15E+07 TCID50/mL

0/3

Adenovirus C1

3.09E+08 TCID50/mL

0/3

Enterovirus 68

1.51E+06 TCID50/mL

0/3

Respiratory Syncytial Virus -A

1.17E+05 TCID50/mL

0/3

Respiratory Syncytial Virus -B

4.57E+06 TCID50/mL

0/3

Human Metapneumovirus (hMPV)

4.17E+05 TCID50/mL

0/3

Rhinovirus 1A

2.20E+07 PFU/mL

0/3

Candida albicans Chlamydia pneumoniae

4.76E+08 CFU/mL

0/3

1.25E+07 IFU/mL

0/3

Haemophilus influenzae

6.97E+08 CFU/mL

0/3

Legionella pneumophila

1.91E+10 CFU/mL

0/3

Streptococcus pneumoniae

1.34E+09 CFU/mL

0/3

Streptococcus pyogenes

2.39E+09 CFU/mL

0/3

Bordetella pertussis

1.96E+10 CFU/mL

0/3

Mycoplasma pneumoniae

2.70E+08 CCU/mL

0/3

Pseudomonas aeruginosa

6.90E+08 CFU/mL

0/3

Streptococcus salivarius Staphylococcus epidermidis

1.20E+08 CFU/mL

0/3

1.40E+08 CFU/mL

0/3

Human Coronavirus 229E

5.62E+04 TCID50/mL

0/3

Human Coronavirus OC43

1.70E+05 TCID50/mL

0/3

Human Coronavirus NL63

1.17E+05 TCID50/mL

0/3

Human Coronavirus HKU1*

5.50E+04 Copies/uL

0/3

Pneumocystis jirovecii

1.00E+08 Nuclei/mL

0/3

SARS-COV-1

3.33E+07 PFU/mL

0/3

MERS-coronavirus

8.90E+05 TCID50/mL

0/3

Mycobacterium tuberculosis

1.15E+08 CFU/mL

0/3

Human Corona HKU1 synthetic RNA was used due to unavailability of virus.

Flu A (# POS / #
Tested)
0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3

Flu B (# POS / #
Tested)
0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3 0/3

Cross Reactive
No No No No No No No No No No No No No No No No No No No No No No No No No No No No No

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Cross-reactivity of Influenza A, Influenza B, and SARS-CoV-2 at high concentrations was evaluated. As shown below, the cross-reactivity testing confirmed that viruses were not cross-reactive at the concentrations tested.

Table 15. Cross Reactive Analysis for Flu A, Flu B, and SARS-CoV-2 Spiked at High Concentrations

Microbial Target

Test Concentration

COVID-19 (POS/#Tested)

FLU A (#POS/#Tested)

Flu B (POS/#Tested)

Cross Reactive

Influenza A/ Hk

9.60E+08 CEID50/mL

0/3

3/3

0/3

No

Influenza A/ Mi

1.00E+09 CEID50/mL

0/3

3/3

0/3

No

Influenza B/ Co

1.60E+08 CEID50/mL

0/3

3/3

No

Influenza B/ Ph

1.10E+09 CEID50/mL

0/3

3/3

No

SARS-COV-2

6.45E+06 TCID50/mL

3/3

0/3

No

(2019-nCoV/USA-

WA1/2020)

b) In Silico In silico analysis was conducted to verify the assay does not cross-react with other high prevalence disease agents and normal or pathogenic flora that are reasonably likely to be encountered in a clinical specimen. Whole genome sequences were downloaded from NCBI. Results are summarized below.
BLAST alignments were found for only two of the species tested: SARS-CoV-1 and Haemophilus influenzae. Since neither of these species had complete primer sets predicted to bind, they are not predicted to have cross reactivity with either primer set for any target analyte. SARS-CoV-1 has > 80% homology on individual primers for SARS-CoV-2 and Candida albicans and Staphylococcus salivarius have > 80% homology on individual primers for Influenza A and were tested and found not to have microbial interference, as shown in Table 18.

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Table 16. Cross-Reactivity BLAST Results

Species
SARS-CoV-1
MERS-CoV Human coronavirus 229E Human coronavirus OC43 Human coronavirus HKU1 Human coronavirus NL63 Adenovirus (e.g. C1 Ad. 71) Human Metapneumovirus (hMPV) Parainfluenza virus 1-4 Influenza A Influenza B Enterovirus (e.g. EV68) Respiratory syncytial virus Rhinovirus Chlamydia pneumoniae Haemophilus influenzae Legionella pneumophila Mycobacterium tuberculosis Streptococcus pneumoniae Streptococcus pyogenes Bordetella pertussis Mycoplasma pneumoniae Pneumocystis jirovecii (PJP) Candida albicans Pseudomonas aeruginosa Staphylococcus epidermidis Staphylococcus salivarius Passes Acceptance Criteria

SARS-CoV-2

Set 1

Set 2

B1c (100%), F2 (100%), F1c (100%) F3 (84%)

N.A.F.

F1c (65%)

N.A.F.

PASS

Influenza A

Set 1

Set 2

N.A.F.

N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. PASS

N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. F1c (71%) N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. LB (86%) N.A.F. N.A.F. F2 (81%) PASS

Set 3
N.A.F.
N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. F3 (77%) N.A.F. N.A.F. N.A.F. PASS

Set 4
N.A.F.
N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. PASS

N.A.F. No Alignment Found. Percentages indicate percent homology to primers with alignments.

Influenza B

Set 1

Set 2

N.A.F.

N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. N.A.F. PASS

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Microbial Interference
a) Competitive Interference by Viral Panel Analytes Competitive microbial interference was tested for SARS-CoV-2, Influenza A, and Influenza B. Each anchor strain was evaluated with 3 sample replicates spiked on a swab at low (3x LoD) concentration and a high level (1E+05 copies/mL) of the anchor strains of the other targets pooled to represent the worst-case scenario. No interference was seen as shown below.

Test Configuration

Table 17. Competitive Microbial Interference Testing Results

Viral Target at 3X LoD
Concentration

Other Viral Targets at High Concentration

COVID-19 Assay Results

Flu A Assay Results

Flu B Assay Results

Competitive Inhibition Present
(Y/N)

Co-spike I

A/HK

B/Ph, SARS-CoV 3/3 positive 3/3 positive 3/3 positive

No

Co-spike II

B/Ph

A/HK, SARS-CoV 3/3 positive 3/3 positive 3/3 positive

No

Co-spike III

SARS-CoV-2

A/HK, B/Ph

3/3 positive 3/3 positive

3/3 positive

No

(2019-nCoV/

USA-WA1/2020)

b) Interference by Other Microorganisms
Microbial interference was evaluated using 29 potential pathogens or commensal organisms. For each organism, nasal swabs were spiked with 35uL of each individual test microorganism and 35uL of a co-spike of SARS-CoV-2, Influenza A (A/Hong Kong/4801/2014) and B (B/Phuket/3073/2013) viral targets diluted in NNSM at 3X LoD final concentration of each virus. The swab was then eluted and run on the Lucira test.
The results showed that no microbial interference was detected.

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Table 18. Microbial Interference by Other Microorganisms Results

Microbial Target
Parainfluenza virus 1 Parainfluenza virus 2 Parainfluenza virus 3 Parainfluenza virus 4 Adenovirus C1 Enterovirus 68 Respiratory Syncytial Virus -A Respiratory Syncytial Virus -B Human Metapneumovirus (hMPV) Rhinovirus 1A Candida albicans Chlamydia pneumoniae Haemophilus influenzae Legionella pneumophila Streptococcus pneumoniae Streptococcus pyogenes Bordetella pertussis Mycoplasma pneumoniae Pseudomonas aeruginosa Streptococcus salivarius Staphylococcus epidermidis Human Coronavirus 229E Human Coronavirus OC43 Human Coronavirus NL63 Human Coronavirus HKU1* Pneumocystis jirovecii SARS-COV-1 MERS-coronavirus Mycobacterium tuberculosis

Test Concentration
1.26E+06 TCID50/mL 1.60E+06 TCID50/mL 8.51E+07 TCID50/mL 1.15E+07 TCID50/mL 3.09E+08 TCID50/mL 1.51E+06 TCID50/mL 1.17E+05 TCID50/mL 4.57E+06 TCID50/mL 4.17E+05 TCID50/mL 2.20E+07 PFU/mL 4.76E+08 CFU/mL 1.25E+07 IFU/mL 6.97E+08 CFU/mL 1.91E+10 CFU/mL 1.34E+09 CFU/mL 2.39E+09 CFU/mL 1.96E+10 CFU/mL 2.70E+08 CCU/mL 6.90E+08 CFU/mL 1.20E+08 CFU/mL 1.40E+08 CFU/mL 5.62E+04 TCID50/mL 1.70E+05 TCID50/mL 1.17E+05 TCID50/mL 5.50E+04 Copies/uL 1.00E+08 Nuclei/mL 3.33E+07 PFU/mL 8.90E+05 TCID50/mL 1.15E+08 CFU/mL

COVID-19 (# POS / #
Tested)
3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3

Flu A (# POS / #
Tested)
3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3

Flu B (# POS / #
Tested)
3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3 3/3

Interference Observed
No No No No No No No No No No No No No No No No No No No No No No No No No No No No No

Human Corona HKU1 synthetic RNA was used due to unavailability of virus.

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Endogenous/Exogenous Interference Substances Studies Endogenous interference studies were conducted to assess potential interference effects on the Lucira assay from substances that may naturally be present in respiratory specimens or artificially introduced onto the nasal swab. 35 µL of the potentially interfering substances listed in the table below was spiked onto the swab at the listed concentrations and evaluated with and without virus spikes:
1. An Influenza A (A/Hong Kong/4801/2014) virus, Influenza B (B/Phuket/3073/2013) virus, and SARS-CoV-2 (2019-nCoV/USA-WA1/2020) virus, all at 3X LoD, were co-spiked to assess Influenza A, Influenza B and SARS-CoV-2 positive performance.
2. NTC devices to evaluate performance in the absence of template.
Substances that yielded 0/3 positive in valid NTC tests and 3/3 positive in valid POS tests were recorded as non-interfering. Invalid tests were repeated until 3 valid results were obtained. As shown in below, none of the substances tested showed interference effects with the Lucira assay.

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Endogenous/ Exogenous Substance
Afrin Original nasal spray Cepacol Choloroseptic Sore Throat Spray Robitussin Mucin, type I-S Nicotine or Tobacco Blood (human) Relenza Tobrex Biotin Zicam Allergy Relief Flonase Nasal Saline spray NeoSynephrine Cold & Sinus Extra Strength Nasacort Mupirocin Tamiflu NeilMed Nasal Gel

Table 19. Endogenous/Exogenous Interference Results

Test Concentration
15% v/v 3 mg/mL
5% v/v

COVID-19 Assay in Presence of
Substance
Pass
Pass
Pass

Flu A Assay in Presence of Substance
Pass
Pass
Pass

Flu B Assay in Presence of Substance
Pass
Pass
Pass

5% v/v

Pass

2.5 mg/mL

Pass

0.03 mg/mL

Pass

5% (v/v)

Pass

5mg/mL

Pass

2.43mg/mL

Pass

3.5 ug/mL

Pass

25% (v/v)

Pass

25% (v/v)

Pass

25% (v/v)

Pass

25% (v/v)

Pass

25% (v/v)

Pass

12mg/mL

Pass

6mg/mL

Pass

1.25% (v/v)

Pass

Interfering (Yes/No)
No No No
No No No No No No No No No No No
No No No No

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Surrogate Sample Testing Study
The Surrogate Sample study compared Lucira COVID-19 & Flu Test performance to that of FDA cleared or authorized comparator methods using samples that were collected in Viral Transport Medium (VTM) and used to prepare contrived specimens for testing. A total of 425 samples were evaluated, and the comparator assays were performed as per the cleared or authorized IFU.
Lucira achieved the following performance using the Lucira COVID-19 & Flu test authorized for use in a POC setting: · SARS-CoV-2: 97.3% positive percent agreement (107/110) and 99.7% negative percent agreement (295/296) · Flu A: 98.4% positive percent agreement (60/61) and 100% negative percent agreement (347/347) · Flu B: 95.3% positive percent agreement (41/43) and 99.7% negative percent agreement (363/364)

Table 20. Surrogate Study Results

Sample Category

Comparator (PCR)

Positive

Negative

Lucira

Success

PPA

N

Total N 95% Wilson CI

Total N

Pos Neg Pos Neg

SARS-CoV-2 107

3

107

97.3%

295

1

295 406

110

92.3% 99.1%

296

Flu A

60

98.4%

347

60

1

0 347 408

61

91.3% 99.7%

347

Flu B

41

95.3%

363

41

2

1

363 407

43

84.5% 98.7%

364

NPA 95% Wilson CI
99.7% 98.1% 99.9%
100% 98.9% 100%
99.7% 98.5% 100%

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Usability and User Comprehension Study
Lucira conducted Human Usability testing among a total sample of 200 healthy, non-symptomatic users to evaluate the ability of various ages, ethnicities, and education levels to successfully run the Lucira COVID-19 & Flu Home Test and interpret test results. 100% (200/200) of users were able to run the test on their own. Participants were each shown six (6) simulated test results and correctly interpreted 99.7% (1194/1200) of results.
Clinical Study
The clinical performance of the Lucira COVID-19 & Flu Home Test was evaluated with symptomatic subjects suspected of COVID-19. Two separate studies were utilized for analyzing performance of the SARS-CoV-2 component of the test.
a) Clinical Study 1 Study 1 was performed using previously authorized Lucira COVID-19 All-In-One Test Kit (SARS-CoV-2 components are in separate chambers and there are no changes to the SARS-CoV-2 primers of this test with respect to the Lucira COVID-19 & Flu Home Test Kit). A total of 101 subjects were enrolled in the study. The subjects independently collected nasal swab samples and ran the test. The results were compared to a high sensitivity molecular FDA Authorized SARS-CoV-2 assay, using samples collected concurrently with those for the Lucira assay that were tested in clinical laboratories.
Table 21. Clinical Study 1 Results

Sample Category

Comparator (PCR)

Positive

Negative

Lucira

N

Pos Neg Pos Neg

Success Total N

PPA 95% Wilson CI

Success Total N

NPA 95% Wilson CI

SARS-CoV-2 48

3

1 49 101

48

94.1%

49

98.0%

51

85.5% 98.4%

50

89.4% 99.9%

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b) Clinical Study 2 Clinical performance of the Lucira COVID-19 & Flu Home Test was evaluated at seven (7) study sites. Prospective nasal samples were collected from patients with signs and symptoms consistent with respiratory infection in the US during the 2022-2023 flu season and were tested at the study sites.
Each patient sample was tested using the Lucira COVID-19 & Flu Home Test and a PCR method as the comparator assay (FDA emergency use authorized SARS-CoV-2 molecular assay and FDA cleared Influenza A&B molecular assay). The Lucira COVID-19 & Flu Home Test was compared against results from the comparator assays.
For prospective specimens, a total of one thousand one hundred sixty-five (1165) subjects were enrolled in the study. One (1) participant withdrew prior to specimen collection and three (3) subjects were excluded due to previous participation. A total of one thousand one hundred sixty-one (1161) samples were evaluated in the performance analysis.
Of the total participants, nine hundred fifty-two (952) participants were evaluated for SARS-CoV-2 results (two hundred-nine (209) samples were assessed ineligible, 195 of which did not have comparator results), one thousand sixty- five (1065) samples were evaluated for Influenza A results (ninety-six (96) samples were assessed as ineligible, 82 of which did not have comparator results) and one thousand sixty-five (1065) samples were evaluated for Influenza B results (ninety-six (96) samples were assessed as ineligible, 82 of which did not have comparator result). Compared to the comparator assay, the Lucira COVID-19 & Flu Home Test demonstrated positive agreement of 88.3% and 90.0% for SARS-CoV-2 and Influenza A, respectively; and negative agreement of 100.0%, 99.3% and 99.9% for SARSCoV-2, Influenza A and Influenza B, respectively. No samples positive for Influenza B were collected during the study.

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Table 22. Clinical Study 2 SARS-CoV-2 Results

SARS-CoV-2 Comparator Test

Positive

Negative

Total

Lucira COVID-19 & Flu Home Test

Positive

83

Negative

111

0

83

858

869

Total

94

858

952

Positive Percent Agreement (PPA)

88.3% (95% CI: 80.2% to 93.3%)

Negative Percent Agreement (NPA)

100.0% (95% CI: 99.6% to 100.0%)

1All false negatives occurred in samples with comparator Ct values of [Target 1] 33.3 – 38.4 and [Target 2] 32.8 – 36.7

Table 23. Clinical Study 2 Influenza A Results

Influenza A Comparator Test

Lucira COVID-19 & Flu Home Test

Positive Negative

Total

Positive Percent Agreement (PPA)

Negative Percent Agreement (NPA)

Positive

Negative

108

7

12

938

120

945

90.0% (95% CI: 83.3% to 94.2%)

99.3% (95% CI: 98.5% to 99.6%)

Total 116 950 1065

Table 24. Clinical Study 2 Influenza B Results

Lucira COVID-19 & Flu Home Test

Positive Negative

Total

Positive Percent Agreement (PPA)

Negative Percent Agreement (NPA)

Influenza B Comparator Test

Positive

Negative

0

1

0

1064

0

1065

N/A

99.9% (95% CI: 99.5% to 100.0%)

Total 1 1064 1065

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Near the Cut-off Evaluation (NTCO)
The Near The Cut-off (NTCO) evaluation study was performed to determine the effects of operator-to-operator variation. Contrived nasal swabs were run by untrained, intended users. The test included 40 well-characterized contrived nasal swab samples: 10 positive contrived samples at 2X LoD for SARS-CoV-2 virus in NNSM, 10 positive contrived samples at 2X LoD for Influenza A virus in NNSM, 10 positive contrived samples at 2X LoD for Influenza B virus in NNSM, and 10 negative contrived samples with NNSM only. This study design tested blinded, contrived swabs prepared by Lucira Health employees and run by ten untrained, intended users. All results in the study were valid and matched the expected results. Overall agreement with expected results was 100% for SARS-CoV-2, Influenza A, Influenza B Positive and Negative samples. The results demonstrate that untrained, intended users are able to use the Lucira COVID-19 & Flu Home Test and obtain the expected results.
Table 25. Summary of NTCO Results by Sample

Sample SARS-CoV-2 Positive
Flu A Positive Flu B Positive
Negative

Percent Agreement (95% CI) 100% (72.2%100%) 100% (72.2%100%) 100% (72.2%100%) 100% (72.2%100%)

(# Successes / # Tested) 10 / 10 10 / 10 10 / 10 10 / 10

Operator #
1 2 3 Total

Table 26. Summary of NTCO Results by Operator and Sample

SARS-CoV-2 Spike

Flu A Spike

Flu B Spike

Negative Spike

(# Positive / # Tested) (# Positive / # Tested) (# Positive / # Tested) (# Positive / # Tested)

3 / 3 4 / 4 3 / 3 10 / 10

4 / 4 3 / 3 3 / 3 10 / 10

0 / 4 0 / 3 0 / 3 0 / 10

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LIMITATIONS
· Performance was evaluated with nasal swab specimens only, using the procedures provided in this instruction. · Failure to follow these procedures may alter test performance. · False negative results may occur if a specimen is improperly collected or handled. · False negative results may occur if inadequate levels of viruses are present in the specimen. · False negative results may occur if the virus mutates in the regions targeted by the test. · The test is a qualitative test and does not provide the quantitative value of detected organism present. · Cross-reactivity with respiratory tract organisms other than those tested in the Analytical Specificity Study may
lead to erroneous results. · This test cannot rule out diseases caused by other bacterial or viral pathogens. · Analyte targets (viral sequences) may persist in vivo, independent of virus viability. Detection of analyte
target(s) does not imply that the corresponding virus(s) are infectious, nor are the causative agents for clinical symptoms. · Positive and negative predictive values are dependent upon prevalence. False negative results are more likely during peak activity when disease prevalence is high and false positive results are more likely during periods of low activity. The performance of the test has not been established in individuals who received nasal administered Influenza vaccine. Individuals who received nasal administered Influenza A vaccine may have positive Influenza A test results for up to three days after vaccination. https://www.cdc.gov/mmwr/preview/ mmwrhtml/rr57e717a1.htm · The performance of this test was established based on the evaluation of a limited number of clinical specimens. Clinical performance has not been established with all circulating variants of SARS- CoV-2 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 home use tests than with laboratory-based molecular tests. This means that there is a higher chance this test will give you a negative result when you have COVID-19. · Performance characteristics for Influenza A was established with clinical specimens collected during the 2022 influenza season when H3N2 was the predominant influenza A virus subtype in circulation. When other influenza A viruses are emerging, performance characteristics may vary. · Influenza B performance was assessed by testing surrogate samples only; it was not assessed by testing natural influenza B positive clinical samples.
TECHNICAL ASSISTANCE
Contact Lucira at customerservice@lucirahealth.com, or call 1-888-LUCIRA-4 (582-4724).
REFERENCES
1. Zhu N, Zhang D, Wang W, et al. A Novel Coronavirus from Patients with Pneumonia in China, 2019. N Engl J Med. 2020;382:727-33. PMID: 31978945.
2. https://www.who.int/emergencies/diseases/novel-coronavirus-2019 3. https://www.cdc.gov/coronavirus/2019-ncov/index.html

Instructions For Use (IFU) INST095 Rev. 6

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TABLE OF SYMBOLS
Product is CE marked.

Product is for single-use only. Do not re-use the same test kit.
Consult the instructions for use.

Product is for in vitro Diagnostic Use.

75 kPa

Total number of IVD tests that can be performed with this IVD is 1.

Caution is necessary when operating the device or control close to where the symbol is placed, or the situation needs operator awareness or operator action in order to avoid undesirable consequences.

30°C 86°F

Store and use product at temperature in the

15°C

range of 15-30°C / 59-86°F.

59°F

Product should not be used if the package has been damaged or opened and that the user should consult the Instructions for Use for additional information.

Use-by date.

80% Store and use the product at relative humidity 10-80%.
10%
The swab is sterilized by ethylene oxide.
STERILE EO

Name and location of the product manufacturer.
Product catalog number.
Product batch code.
106 kPa Store and use the product at atmospheric pressure in the range of 75-106 kPa.
Batteries within the test unit should be disposed of separately from household waste and recycled. Applies in the European Union only. Test unit should be disposed of separately from household waste and recycled. Applies in the European Union only.
Type BF applied part.

Lucira Health, Inc. 1315 63rd Street, Emeryville, CA 94608 United States
Instructions For Use (IFU) INST095 Rev. 6

Covered by one or more of US Patents 10,146,909, 10,253,357 and other pending US and International Patents.
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