Lucira COVID-19 check it Test Kit Instructions

Lucira COVID-19 check it Test Kit

Lucira Tradmarked Check (checkmark) It Covid-19 Test Kit

Instructions for Use

• For Use under Emergency Use Authorization (EUA) only
• For in vitro Diagnostic Use Only
• For Use with Self-collected Nasal Swab Specimens in individuals aged 14 and older
• For Over-the-Counter (OTC) Use

Intended Use

The Lucira CHECKIT COVID-19 Test Kit is a single-use test kit intended for the qualitative detection of nucleic acid from the novel coronavirus SARS- CoV-2 that causes COVID-19. This test is for nonprescription home use with self-collected anterior nasal (nasal) swab specimens in individuals aged 14 years and older (self-collected) or individuals ≥2 years (collected by an adult) with or without symptoms or other epidemiological reasons to suspect COVID-19.
This test is similar to a PCR test in that it utilizes a molecular amplification technology for the detection of SARS-CoV-2 viral RNA. SARS-CoV-2 viral RNA is generally detectable in anterior nasal swab specimens during the acute phase of infection. Positive results indicate the presence of viral RNA, 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. Negative results are presumptive and confirmation with a molecular assay performed in a laboratory, if necessary for patient management, may be performed. Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for treatment
or management decisions for the individual, including infection control decisions. Negative 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. It is important to consult healthcare providers to discuss test results and if additional testing is necessary.
The Lucira CHECK IT COVID-19 Test Kit is for use only under the Food and Drug Administration’s Emergency Use Authorization. Test results can be reported through the LUCI secure web portal, 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 the CDC.

Summary and Explanation of the Test
Coronavirus disease 2019 (COVID-19) is a pandemic respiratory illness caused by a novel human coronavirus, first isolated in China and named SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) by the World Health Organization.1-3 SARS-CoV-2 has spread globally and can cause mild to severe respiratory illness including death. The Lucira CHECKIT COVID-19 Test kit is a single use molecular diagnostic test kit for the qualitative detection of SARS-CoV-2 RNA from nasal swab samples and can be used by individuals with or without COVID-19 symptoms. Each test kit contains all the components required to perform one (1) COVID-19 test.

Principles of the Procedures
The Lucira CHECK IT COVID-19 Test Kit utilizes RT-LAMP technology to detect RNA of the N gene for SARS-CoV-2. RT-LAMP is a molecular amplification technology similar to PCR. 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 chambers that target SARS-CoV-2 RNA (two non- overlapping regions of the N gene), a positive internal control (PIC), and a lysis internal control (LIC).
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 1 hour after the test has finished running.

PRECAUTIONS – GENERAL

• For FDA Emergency Use Authorization (EUA) only
• For in vitro diagnostic use.
• This OTC test kit has not been FDA cleared or approved, but has been authorized for emergency use by FDA under an EUA.
• This OTC test kit has been authorized only for the testing of nasal swabs for detection of nucleic acid from SARS-CoV-2, not for any other viruses or pathogens.
• The emergency use of this OTC test kit 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 revoked sooner.
• Leave kit components sealed in foil pouches until just before use.
• It is important to follow instructions for use. Proper use is essential for correct results.
• Do not insert batteries into test unit until ready to perform test.
• Do not touch swab tip when handling swab sample.
• Do not use any kit components with visible damage.
• Use the Product only as specified, or the protection supplied by the Product can be compromised.
• Do not use kit components after their expiration date.
• Choose a level location to do this test where test can sit undisturbed for 30 minutes.
• All kit components are single use items. Do not attempt to use the same test kit a second time.
• After use, place test unit in disposal bag and dispose all test kit materials in trash.

SECTION A: Reagents and Materials

Lucira™ CHECKIT COVID-19 Test Kit contents:

• Package Insert
• Nasal Swab: one sterile 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 RNA;
• Batteries: two AA batteries for the Test Unit; and
• Plastic disposal bag to dispose of the test kit after use.

NOTE: It is important to follow instructions for use. Proper use is essential for correct results.

STORAGE AND HANDLING

• Test kits must always be stored at an ambient temperature (15-30°C / 59-86°F).
• Do not open kit components sealed in foil pouches until immediately before use.
• Do not reuse kit components.

Section B – Directions for running the Lucira COVID-19 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.
• Do not discard box so it can be used for LUCI reporting.
• 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. 

Set Up Your 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

Note: Keep vial away from children. Avoid contact with eyes and skin. If contact occurs, rinse with water. If irritation persists, seek medical attention.

Swab Both Nostrils
For this test to work properly, it is important you 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/patient nostril and you meet resistance.
• 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.
  Rotate 5x in BOTH nostrils.
  Make sure to roll around inside walls to collect a good sample. 

Adults must swab children ages 2-13.

Stir Swab and Run Test

• Insert swab into the sample vial until it touches the bottom.
• Mix sample by stirring around the sample vial 15 times.
• Discard swab.
• Snap cap closed and press vial down into test unit until it clicks.
• Ready light will start blinking when test is running. 

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.

Read and Report Result
Done light will display when test is ready in 30 minutes.

Next, please use Lucira’s secure LUCI portal to receive a verified test result on your phone and transmit your result to public health agencies. To get started, please text the word ‘LUCI’ to 44544.

If the test is POSITIVE
It is very likely you have COVID-19 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 you test NEGATIVE
A negative result means the virus that causes COVID-19 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. This means you could possibly still have COVID-19 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 a 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.

Dispose of Test Kit
After test is completed, place the test unit in plastic disposal bag and dispose all test kit materials in trash.

Section C – Test Unit Display Result

When the test is complete, the results are clearly displayed on the Test Unit.

Section D – Quality Control Testing for OTC Settings

The Lucira CHECKIT is a single-use test kit and does not require external run controls (ERCs).

Section E – Conditions of Authorization for OTC Use

The Lucira COVID-19 All-In-One Test Kit Letter of Authorization, along with the authorized Fact Sheet for Healthcare Providers and authorized labeling are available on the FDA website:
https://www.fda.gov/medical-devices/coronavirus-disease-2019-covid-19 -emergency-use-authorizations-medical-devices/in-vitro-diagnostics-euas- molecular-diagnostic-tests-sars-cov-2

PERFORMANCE CHARACTERISTICS

1. Limit of Detection (LoD) – Analytical Sensitivity
   Quantified heat-inactivated SARS-CoV-2 virus was serially diluted in Natural Nasal Swab Matrix (NNSM), 35 µL was pipetted onto a fresh, unused nasal swab, and run on two device lots. The LoD for the device was determined by testing three (3) target concentrations 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. The LoD for each lot was separately determined as the lowest concentration of genome copy equivalents per swab that yielded greater than 95% positive results. The preliminary LoD for the device was defined as the highest LoD of the two lots. Genome equivalent

/ Swab

(per reaction)

| ****

Positive/Total Valid

| ****

Percent Positive

---|---|---
 | Preliminary Lot 1| Preliminary Lot 2| Confirmatory| Preliminary Lot 1| Preliminary Lot 2| Confirmatory
2700*| 20/21| 20/21| 20/20| 95.2%| 95.2%| 100%
1350| 18/21| 20/21| —| 85.7%| 95.2%| —
675| 16/21| 15/21| —| 76.2%| 71.4%| —

*2700 cp/swab is determined as LOD and is equivalent to 900 cp/mL of VTM assuming 100% elution of the swab in 3 mL of VTM. The LoD for Lot 1 was determined to be 2700 copies per swab, while Lot 2 was 1350 genome equivalents per swab. As such, 2700 copies per swab was reported as the preliminary LoD.
The LoD was confirmed by testing 20 replicates at the preliminary LoD concentration on a single lot. All 20 of 20 devices at this concentration were positive.

2. Inclusivity (analytical sensitivity)
   Inclusivity of the Lucira CHECKIT COVID-19 Test Kit was demonstrated by in- silico reactivity of the assay against all publicly available SARS-CoV-2 strains using the assay’s primers and probes. A total of 28,555 SARS-CoV-2 sequences were downloaded from the Global Initiative on Sharing All Influenza Data (GISAID, https://www.gisaid.org) database, and trimmed from whole genomes to 2.4-3.6kb windows covering the target regions. All 28,555 (100%) of the downloaded valid sequences were found to be reactive to at least one primer set of the assay.
   Regarding the number of SNPs, none of the sequences had 3 or more mismatches to either primer set. 95.98% and 98.35% of the 28,555 sequences evaluated had 100% identity to Set 1 and Set 2, respectively. 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).
   Lucira performs monthly surveillance of emerging SARS-CoV-2 strains by periodically evaluating in silico reactivity against sequence databases to ensure that emerging strains of SARS-CoV-2 are reactive to the Lucira CHECK IT COVID-19 Test Kit. As of March 2, 2021, all of the sequences from variants of concern as identified by CDC were found to be reactive to at least one primer set of the assay. The analysis included 16,572 sequences of B.1.1.7, 261 sequences of B.1.351, 195 sequences of P.1, and 2,737 sequences of B.1.427/B.1.429. A Technical Brief related to Lucira’s most current analysis is available on Lucira’s website.

3. Cross-reactivity (Analytical Specificity)
   

4. Analytical specificity of the Lucira CHECK IT COVID-19 Test Kit was demonstrated by testing cross-reactivity with and interference from other organisms as well as endogenous substances.
   a. Cross-reactivity (organisms tested in the absence of SARS-CoV-2) The specificity of the assay was evaluated in cross-reactivity testing using 33 commensal organisms, including 13 bacteria/fungi, and 20 viruses. The cross- reactivity testing confirmed that none of the 33 organisms were cross-reactive with the Lucira CHECK IT COVID-19 Test Kit at the concentrations tested. ****

Organism Tested

| Concentration Tested (per mL of spike)| Number Pos / # Tested| Cross Reactive
---|---|---|---
Bordetella pertussis| 1.34E+10 CFU/mL| 0/3| No
Candida albicans| 6.27E+08 CFU/mL| 0/3| No
Chlamydia pneumoniae| 1.25E+07 IFU/mL| 0/3| No
Haemophilus influenzae| 2.27E+09 CFU/mL| 0/3| No
Legionella pneumophila| 6.82E+09 CFU/mL| 0/3| No
Mycobacterium tuberculosis| 6.86E+07 CFU/mL| 0/3| No
Mycoplasma pneumoniae| 3.98E+07 CCU/mL| 0/3| No
Pseudomonas aeruginosa| 6.64E+07 CFU/mL| 0/3| No
Pneumocystis jirovecii| 1.0E+08 nuclei/mL| 0/3| No
Staphylococcus epidermidis| 1.32E+08 CFU/mL| 0/3| No
Streptococcus pneumoniae| 4.73E+08 CFU/mL| 0/3| No
Streptococcus pyogenes (Group A)| 4.30E+09 CFU/mL| 0/3| No
Staphylococcus salivarius| 1.2E+08 CFU/mL| 0/3| No
---|---|---|---
Adenovirus 1| 1.02E+08 TCID50/mL| 0/3| No
Adenovirus 7| 1.02E+08 TCID50/mL| 0/3| No
Coronavirus 229E| 1.26E+06 TCID50/mL| 0/3| No
Coronavirus NL63| 1.1E+06 TCID50/mL| 0/3| No
Coronavirus OC43| 1E+6.18 TCID50/mL| 0/3| No
Enterovirus/Coxsackievirus B4| 6.61E+06 TCID50/mL| 0/3| No
Enterovirus Type 68| 9.55E+06 TCID50/mL| 0/3| No
Enterovirus Type 71| 4.17E+05 TCID50/mL| 0/3| No
Human metapneumovirus| 1.0E+06 TCID50/mL| 0/3| No
Influenza A (pH1N1)| 1.58E+07 EID50/mL| 0/3| No
Influenza B (Yamagata)| 1.1E+08 CEID50/mL| 0/3| No
MERS Coronavirus| 8.9E+05 TCID50/mL| 0/3| No
Parainfluenza 1| 8.9E+06 TCID50/mL| 0/3| No
Parainfluenza 2| 1.0E+08 TCID50/mL| 0/3*| No
Parainfluenza 3| 6.6E+06 TCID50/mL| 0/3| No
Parainfluenza 4A| 4.57E+06 TCID50/mL| 0/3| No
Respiratory Syncytial Virus Type A| 3.16E+06 TCID50/mL| 0/3| No
Respiratory Syncytial Virus Type B| 1.26E+06 TCID50/mL| 0/3| No
Rhinovirus 1A| 1.58E+08 TCID50/mL| 0/3| No
SARS Coronavirus (SARS-CoV-1)| 1.0E+08 PFU/mL| 0/3| No

• One replicate repeated with another device due to an initial Invalid result.
  In addition, 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. BLAST alignments showed that only SARS-CoV-1 has > 80% homology on individual primers.

Microbial Interference Studies (organisms tested in the presence of SARS- CoV-2)Based on cross-reactivity testing and in silico sequence alignment reported above, SARS-CoV-1 was identified as the only microorganism that may potentially interfere with the performance of the Lucira assay for SARS-CoV-2. In addition to SARS-CoV-1, Influenza (Flu) A, Flu B, respiratory syncytial virus (RSV) A, and RSV B were tested to verify that these high prevalence pathogenic microorganisms do not interfere with the detection of SARS-CoV-2 in the Lucira assay. In order to evaluate potential interference, swabs were co- spiked with 35 µL of the potentially interfering microorganisms, at the concentrations listed in Table 4 below, with SARS-CoV-2 (at just less than 3 × LOD at 8.08 x 103 genome equivalents / swab) diluted in NNSM. Three positive spikes of only SARS-CoV-2 viral template and three NTC devices were run as controls.  The results showed that no microbial interference was detected for any of the microorganisms tested. Devices spiked with SARS-CoV-2 at 8.08 x 103 genome copies/swab (less than 3 × LOD) were positive, regardless of the presence of SARS-CoV-1, Flu A, Flu B, RSV A, or RSV B. All controls gave the expected results as well. As already shown in Table 2 above, these five microorganisms do not cross-react with the assay. Therefore, these microorganisms tested are unlikely to interfere with the performance of the assay.

Organism Tested

| ****

Concentration Tested (35 µL spiked per swab)

| Number Positive / Valid Tests| Lucira CHECK ü IT COVID-19 Test Kit
---|---|---|---
SARS-CoV-1| 1.00E+08 PFU / mL| 3 / 3| No
Influenza A pH1N1 A/Michigan/45/2015| 1.58E+07 EID50 / mL| 3 / 3| No
Influenza B| ****

1.10E+08 CEID50 / mL

| ****

3 / 3

| ****

No

Respiratory Syncytial| 3.16E+06 TCID50 / mL| 3 / 3| No
Respiratory Syncytial| 1.26E+06 TCID50 / mL| 3 / 3| No

Endogenous Interference Substances Studies
Endogenous interference studies were conducted to assess potential interference effects on the Lucira CHECKIT COVID-19 Test Kit from substances that may naturally be present in respiratory specimens or artificially introduced onto the nasal swab, including common household items that may be present in the testing environment.
Fifteen (15) interfering substances were tested, including 12 substances that are naturally present or artificially introduced into the nasal cavity, which were tested at the highest medically relevant concentration, and 3 substances that may be present within the testing environment, which were tested at their highest expected concentrations in the environment. These potentially interfering substances were tested in NNSM spiked with SARS-CoV-2 virus at less than 3× the LoD, and without spiking. None of the 15 substances tested showed interference effects with the Lucira CHECKIT COVID-19 Test Kit.

Substance

| ****

Concentratio n Tested

| NTC COVID-19

Positive Results / Valid Devices

| POS COVID-19

Positive Results / Valid Devices

| ****

Interfering Substance

---|---|---|---|---
PBS (control)| undiluted| 0/3| 3/3| No
Afrin| undiluted| 0/3| 3/3| No
Blood| 1.0 % v/v| 0/3| 3/3| No
Chloroseptic Max| 20% w/v| 0/3| 3/3| No
Flonase Allergy Relief| undiluted| 0/3| 3/3| No
Mucin| 0.5% w/v| 0/3| 3/3| No
Method All-Purpose| 5% v/v| 0/3*| 3/3| No
Mupirocin| 4.3 mg/ mL| 0/3| 3/3| No
NeoSynephrine Cold| ****

20% v/v

| ****

0/3

| ****

3/3

| ****

No

Relenza| 10 mg/mL| 0/3| 3/3| No
Saline| undiluted| 0/3| 3/3| No
Seventh Generation| 5% v/v| 0/3| 3/3| No
Softsoap Moisturizing| 5% v/v| 0/3**| 3/3| No
Tamiflu| 6 mg/mL| 0/3| 3/3| No
Tobramycin| 2.5 mg/ mL| 0/3| 3/3| No
Zicam Allergy Relief| undiluted| 0/3| 3/3| No

1. NTC condition involved testing the potentially interfering substances in NNSM.
2. POS condition involved testing the potentially interfering substances in NNSM spiked with SARS-CoV-2 virus at less than 3× the LoD.

• One replicate repeated with another device due to an initial Invalid result. **Two replicates repeated with additional devices due to initial Invalid results.

  5. Near the Cutoff Study
     To demonstrate the reproducibility of the Lucira CHECKIT COVID-19 Test Kit with analyte concentrations near the cutoff, a study was conducted with 20 contrived nasal swabs run by untrained, intended users. The test included 20 well-characterized contrived nasal swab samples: 10 positive contrived samples at 2×LOD for SARS-CoV-2 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 10 untrained, intended users with various level of experience.
     All results in the study were valid and matched the expected results. Overall agreement with expected results was 100% for both SARS-CoV-2 Positive and SARS-CoV-2 Negative samples. The results demonstrate that untrained, intended users are able to use the Lucira CHECKIT COVID-19 Test Kit and obtain the expected results.

  6. Flex Studies
     To assess the robustness of the Lucira CHECKIT COVID-19 Test Kit, the company has conducted the following flex studies to test the assay’s performance under conditions that may be evaluated as potential user errors and anticipated environmental stresses.
     a. Sample Mixing
     The study was conducted to evaluate the impact of incomplete or excessive swab mixing in the sample vial on performance of the Lucira CHECKIT COVID-19 Test Kit. For each condition, 3 devices were run using the positive samples. All devices tested with positive samples returned a positive result, demonstrating that the assay is robust to incomplete or excessive swab mixing conditions.

     ---

Flex Condition

| Positive Spike (#Positive/ # Valid
---|---
0 Swirls (12 Second Dip)| 3/3
5 Swirls| 3/3
10 Swirls| 3/3
40 Swirls| 3/3

Contaminating Substance
The study was conducted to evaluate the impact on device performance from hand sanitizer or hand lotion that might be transferred to the swab head from a user’s hand. All devices displayed expected results.

Flex Condition

| Positive Spike (# Positive / # Valid)| NTC Condition

(# Positive / # Valid)

---|---|---
Bare Hands| 3/3| 0/3
Hand Lotion| 3/3| 0/3
Hand Sanitizer| 3/3| 0/3

Battery Removal
The study was conducted to evaluate the impact of removal of a battery, as well as sequential removal and re-insertion of a battery after a device has started running, on the Lucira CHECK  IT COVID-19 Test Kit performance. The results showed that when battery was removed, the device turned off; and that if the battery was re-inserted, the device displayed an “Invalid” result, as designed.
Device Movement
The study was conducted to evaluate the impact of vertical and horizontal device movement and tilting of the device while the device is running on the Lucira CHECK IT COVID-19 Test Kit performance. Each device underwent vertical movement at 0 minute to simulate someone picking up the device immediately after starting the device, vertical and horizontal movements at 12 minutes to simulate picking up and moving the test to another room during a test run, and tilting at 12 minutes to simulate the user picking up and tilting the device to observe the display.

Movement

| Vertical Movement| 1 foot| 0 minutes / 12 minutes
Horizontal Movement| 20 feet| 12 minutes

Tilting

| 90° perpendicular to ground

(device tilted from long axis horizontal to long axis vertical)

| ****

12 minutes *

The movements at 12 minutes were performed in the following order: Vertical Movement, Horizontal Movement, Tilting. All devices displayed expected results following the movement conditions.

Unlevel Surface
The study was conducted to evaluate the impact of device placement on an angled work surface on the Lucira CHECKIT COVID-19 Test Kit performance. Devices were run on a custom fixture with slope. Each sample and orientation were tested in triplicate, with devices kept in their position for the full duration of the run.
All devices run following this test protocol displayed expected results, satisfying the acceptance criteria.

Storage of Packaged Devices
The study was conducted to evaluate the tolerance of the Lucira CHECKIT COVID-19 Test Kit to short-term storage at low and high temperatures outside the recommended temperature storage range. Device performance was assessed within a storage temperature range of 5°C to 45°C for up to 3 days. For each condition (temperature and duration combination), 3 devices were tested per sample type. All tested devices displayed expected results following the low and high temperatures.

Condition

| ****

Description

| Positive Spike (# Positive / # Valid)| NTC Condition (# Positive / # Valid)
---|---|---|---
Storage Temperature and Time| 5°C – 24 hours| 3/3| 0/3
Storage Temperature and Time| 5°C – 72 hours| 3/3| 0/3
Storage Temperature and Time| 45°C – 24 hours| 3/3| 0/3
Storage Temperature and Time| 45°C – 72 hours| 3/3| 0/3

Shipping and/or storage of kits in extreme conditions
The Lucira CHECKIT COVID-19 Test Kits, in a shipper without temperature- control assistance, underwent a climatic simulation per ASTM D4332-14 and a shipping simulation per ASTM D4169-16 Distribution Cycle 13 to demonstrate the stability of packaged test kits under multiple temperature and humidity conditions expected during shipments. Shipping testing was done with four packages containing 24 test kits each packaged in a shipper (single-corrugated ECT>44 cardboard box from Packaging Corporation of America) with no additional temperature-control assistance. These four packages were subjected to testing outlined in the ASTM D4332-14 Standard Practice for Conditioning Containers, Packages, or Packaging Components for Testing as shown in Table 10 below.
Table 10: Temperature and Humidity Conditioning per ASTM D4332-14

Shipper

| Temperature/Humidity Conditions| Total           Conditioning Time
---|---|---
+60ºC/15%| -30ºC| +40ºC/90%
S1| X|  |  | 24 hours
S2|  | X|  | 24 hours
S3|  |  | X| 24 hours
S4| X| X| X| 72 hours

Kits from these shippers were tested with known positive and negative samples to evaluate any functional impacts of the climactic and shipping testing. Eight (8) kits from the corners of each shipper were tested and the kits passed the acceptance criteria of successfully classifying four known positive samples, at 3 × LoD in NNSM, and four known negative samples (PBS in NNSM matrix) per shipper. One kit from Shipper S4 generated an invalid result and was successfully retested with an additional kit from the shipper. These functional results demonstrate that there was no impact to the performance of the kits when exposed to several temperature and humidity conditions, including below freezing.

Operating Temperature and Humidity Extremes
The study was conducted to evaluate the tolerance of the Lucira CHECKIT COVID-19 Test Kit to both temperature and humidity extremes which may be encountered during normal operation of the device by end users. Device performance was assessed within extreme temperature and humidity combinations, with temperature ranging from of 5°C to 45°C and humidity ranging from 5% to 95%.
All devices displayed expected results under the tested environment conditions. The 5°C and 45°C conditions resulted in “Invalid” results for all devices tested as they were out of the temperature range upon the device waking up at battery insertion.

Condition| Positive Spike

(# Positive / # Valid)

| NTC Condition

(# Positive / # Valid)

---|---|---
10°C, Low Humidity| 3/3| 0/3
35°C, Low Humidity| 3/3| 0/3
45°C, Ambient Humidity| 0/0| 0/0
20°C, Ambient Humidity| 3/3| 0/3
35°C, High Humidity| 3/3| 0/3
10°C, High Humidity| 3/3| 0/3
5°C, Ambient Humidity| 0/0| 0/0

All test units tested were invalid by design.

Human Usability Study
Lucira conducted Human Usability testing among a total sample of 398 healthy, non-symptomatic users in two locations to evaluate the ability of various ages, ethnicities and education levels to successfully run the Lucira CHECKIT COVID-19 Test Kit.
For all of Lucira’s Usability studies, participants were recruited to come to a research facility and try the Lucira All-In-One Test Kit on their own using only the Package Inserts provided in the test kit box. Subjects were observed through a one-way mirror and videoed so that complete records of their test kit use experience, including nasal self-collection, could be documented and verified. Post-use interviews to assess test kit ease of use and instructions clarity were entered electronically in real time.
All users of all ages were able to successfully start the Lucira test running. Notably, 96% of all users aged 14 and above were able to successfully start the test running and ready light blinking on the first try and without having to look back at the Package Insert instructions. The remaining were all able to successfully start the test running with one look back at the instructions. 97% of users rating their experience as “Easier or About What They Expected,” with 59% of users rated their test experience as “Easier Than What They Expected.” Nearly 100% of users rated the overall test kit instructions as sufficient to be able to understand how to perform the test. After using the Lucira CHECKIT COVID-19 Test Kit, 100% of users across all age cohorts expressed confidence they could run the test at home on their own. All users were able to interpret the test results correctly. 99% of adults aged 18 and older and 95% of teens aged 14-17 collected sample in both nostrils. Among the parent collection sample, 98% were able to collect a sample in both their child’s nostrils. 92% of adults aged 18 and older had a swabbing time of at least 11 seconds and a median swabbing time of 18 seconds. 92% of teens aged 14-17 had a swabbing time of 9 seconds and a median swabbing time of 17 seconds. 90% of parent assisted collections had a swabbing time of 9 seconds and a median swabbing time of 13 seconds.
Clinical Evaluation:
Lucira has conducted two Community Testing Studies to establish the clinical performance of the Lucira CHECKIT COVID-19 Test Kit among individuals both with and without COVID-19 symptoms, respectively. In the symptomatic study, subjects were enrolled who were: 1) experiencing at least three of the ten CDC COVID-19 symptoms and including at least one symptom being cough, shortness of breath or new loss of taste or smell; 2) known COVID positive in the past 14 days and experiencing at least one CDC COVID-19 symptom; or 3) currently experiencing fever or self-reported fever of 100 degrees or higher in past 48 hours and one additional CDC symptom. In the asymptomatic study, only subjects who were not experiencing any of the CDC COVID-19 symptoms at the time of specimen collection and testing were enrolled. In both of these studies, subjects were tested outside their home in a real-world setting. All subjects independently self-collected Lucira nasal swab samples and successfully ran the test. After performing the Lucira test, all subjects self-collected a second nasal swab sample which was tested on a FDA authorized high sensitivity SARS-CoV-2 PCR test. Results were compared between the Lucira test and the PCR test run in clinical laboratories.
In the two Community Testing studies that included a total of 404 individuals with and without COVID-19 symptoms, the Lucira test achieved 98% (267/272) negative percent agreement (NPA) when compared to a FDA authorized high sensitivity SARS-CoV-2 PCR test. Positive percent agreement (PPA) in the symptomatic study was 94% and 90% in asymptomatic study. Total PPA was 92% (121/132) across all subjects and included 10 samples with very low levels of virus (>37.5 Ct) as shown in Table 14 below.

Subject Type

| Lucira Community Tes�ng Study Summary Clinical Performance

Vs High Sensi�vity FDA EUA SARS-CoV-2 PCR Test

---|---
PPA| NPA

Symptoma�c

| 94% (48/51)| 98% (49/50)

Asymptoma�c

| 90% (73/81)| 98% (218/222)

Total

| 92% (121/132)| 98% (267/272)

*Seven (7) of the 11 discrepant samples had high Ct values (>37.5) when tested by the comparator assay.

Lucira Community Tes�ng Studies

| A High Sensi�vity FDA Authorize d SARS-CoV-2 PCR T est
---|---
Posi�ve| Nega�ve| Total
Positive| 121| 5| 126
Negative| 11| 267| 278
Total| 132| 272| 404

• Positive Percent Agreement (PPA)
• Negative Percent Agreement (NPA)
• 7 invalid result (1.73% invalid rate); 2 retests (0.49% retest rate)
• 91.7% (95%CI: 85.6%-95.8%)
• 98.2% (95%CI: 95.8%-99.4%)

*Seven (7) of the 11 discrepant samples had high Ct values (>37.5) when tested by the comparator assay.
In the graph below of Ct values of reference positives samples from the Community Testing studies, BLUE color is used to show that the Lucira test result matched the comparator positive result. GREY color is used to show that the Lucira test result did not match the comparator reference test result. Nearly all of the GREY bars occurred in samples where there were very low levels of virus detected by the comparison test. Ct values are shown in ascending order.

Table 13. Lucira vs. a High Sensitivity Molecular FDA Authorized SARS-CoV-2 PCR Assay Positive Percent Agreement Summary

*Seven (7) of the 11 discrepant samples had high Ct values (>37.5) when tested by the comparator assay.
The symptomatic Community Testing study enrolled a total of 101 subjects. When compared to a FDA authorized high sensitivity SARS-CoV-2 PCR test, the total PPA across all samples was 94.1% (48/51), or 96.0% (48/50) with discrepancy analysis using the Roche cobas SARS-CoV-2 test. All three of the discrepant samples had high Ct values (>37.5) when tested by the comparator assay. There was 1 invalid result (0.99%) and 2 retests (1.98%) in this study.
Table 16. Clinical Study Summary Results – Symptomatic

A High Sensi�vity FDA

Community Tes�ng Study                           Authorized SARS-CoV-2 PCR Assay

Lucira 07A-CLI-006| Posi�ve| Nega�ve| Total
Posi�ve| 48| 1| 49
Nega�ve| 3*| 49| 49
Total| 51| 50| 101

Positive Percent Agreement (PPA)
Negative Percent Agreement (NPA)
*All 3 False Negatives occurred in parents with reference test Ct values of >37.5 1 invalid result (0.99% invalid rate); 2 retests (1.98% retest rate) The asymptomatic Community Testing study enrolled a total of 303 subjects. When compared to a high sensitivity molecular FDA Authorized SARS-CoV-2 assay, the total PPA across all samples was 90.1% (73/81). Four of the 8 discrepant samples had high Ct values (>37.5). There were 6 invalids (1.98%) in this study.

Community Tes�ng Study Lucira 07A-CLI-007

| A High Sensi�vity FDA Authorized SARS-CoV-2 PCR Test
---|---
Posi�ve| Nega�ve| Total
Posi�ve| 73| 4| 77
Nega�ve| 8*| 218| 226
Total| 81| 222| 303

LIMITATIONS

• Performance was evaluated with 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 the 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 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.
• The performance of this test was established based on the evaluation of a limited number of clinical specimens. 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.

TECHNICAL ASSISTANCE
Contact Lucira at customerservice@lucirahealth.com, or call 888-582-4724.

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 

TABLE OF SYMBOLS

| This symbol indicates that the product is for single use only. It is not to be re-used.
---|---
| This symbol indicates that you should consult the instructions for use.
| This symbol indicates that the product has a temperature limitation.
| This symbol indicates the use-by date.
| This symbol indicates the product batch code.
| This symbol indicates the name and location of the product manufacturer.
| This symbol indicates the product’s catalog number.
| For In Vitro Diagnostic Use.

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

References

• Coronavirus Disease 2019 (COVID-19) | CDC
• GISAID - gisaid.org
• Coronavirus disease (COVID-19)

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