ACCURACY & DURABILITY - LATERAL FLOW - SARS-CoV-2 RAPID ANTIGEN

Overseas Production

SARS-CoV-2 rapid Antigen test

OROPHARYNGEAL

RESULTS IN 20 MINUTES

Asymptomatic Identified

Specificity

Sensitivity

CT values to 30

SARS-CoV-2 rapid Antigen test

ANTERIOR NARES

RESULTS IN 20 MINUTES

OROPHARYNGEAL - Oral Administration - Swabs for Children

=> ongoing worldwide clinical trials performed on over 500 subjects from 2020-2021

=> 0% False Positives accomplished with 390 negative patient samples

=> 0% Cross Reactivity accomplished with 44 common viruses including SARS-CoV-1, interfering substances and bacteria

=> Withstanding Extreme Heat & Freezing Environments

=> Extended shelf-life

Backed by Robust Clinical Studies

The SARS-CoV-2 COVID-19 Antigen test is a lateral flow immunochromatographic assay intended for the qualitative detection of the nucleocapsid protein antigen from SARS-CoV-2 in Oral, nasopharyngeal or anterior nasal swab specimens directly collected from individuals who are either suspected of COVID-19 by their healthcare provider within first five days of symptom onset. Testing is limited to qualified and trained clinical laboratory personnel specifically instructed and trained in the techniques of collection and interpretation by Spectrum Diagnostics/Inner Health laboratory certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, that meets the requirements to perform moderate, high or waived complexity tests operating under CLIA Certificate of Waiver, Certificate of Compliance, or Certificate of Accreditation.

The LDT COVID-19 Antigen is intended for use in clinical settings and operated by healthcare professionals or trained users specifically instructed in the use of the COVID-19 Antigen Test Kit and proper infection control procedures. Testing with this Laboratory Developed Test, LDT is intended for use by trained clinical laboratory staff specifically instructed and trained in the techniques of sample collection under medical or CLIA license. Although results can be read onsite, they are not official until uploaded to the Test platform and validated by the Lab Director.

Results are for the identification of the SARS-CoV-2 nucleocapsid protein antigen. The antigen is generally detectable in nasopharyngeal or anterior nasal swab specimens during the acute phase of infection. Positive results indicate the presence of viral antigens, but the clinical correlation with patient history and other diagnostic information is necessary to determine infection status. Positive results do not rule out a bacterial infection or co-infection with other viruses. The agent detected may not be the definite cause of disease. Laboratories within the United States and its territories are required to report all results to the appropriate public health authorities.

Negative results should be treated as presumptive and confirmation with a molecular assay, if necessary, for patient management, may be performed. Negative results do not rule out SARS-CoV-2 infection and should not be used as the sole basis for treatment or patient management decisions, including infection control decisions. Negative results should be considered in the context of a patient’s recent exposures, history, and the presence of clinical signs and symptoms consistent with COVID-19.

The SARS-CoV-2 COVID-19 Antigen is a rapid – 20 minutes chromatographic immunoassay for the direct detection of the presence or absence of SARS-CoV-2 antigens in the respiratory specimens taken from patients with signs and symptoms who are suspected of COVID-19, or taken from asymptomatic individuals, as described in the intended use.

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

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

Positive results for asymptomatic patients

Cumulative RT-PCR Pos(+)

Cumulative Covid-19 Antigen Pos (+)

PPA

95% Confidence

22

22

100.0%

98.81 – 99.86%

SARS-CoV-2 COVID-19 Antigen nasal clinical performance within 9 days of symptom onset against the comparator method

Analytical Sensitivity: Limit of Detection (LoD):
The LoD for direct swab was established using heat-inactivated SARS-CoV-2 isolate USA-WA1/2020 (catalogue number 0810587CFHI) – Lot number 325828 – Exp date 2024/01/22. The strain was spiked into the pooled human nasal swab matrix obtained from multiple healthy volunteers eluted in VTM and confirmed as SARS-CoV-2 negative by RT-PCR to prepare positive samples. The estimated LoD found from the initial two-fold serial dilution test was confirmed by testing 20 replicates. The confirmed LoD for direct swab was 2.5 x 102 TCID50/ml.

Analytical Specificity Cross Reactivity (Exclusivity) and Microbial Interference:
The potential cross-reactivity (exclusivity) of a panel of common organisms was evaluated with SARS-CoV-2 negative samples using the SARS-CoV-2 COVID-19 Antigen test. Potential microbial interference was evaluated with samples containing heat-inactivated SARS-CoV-2 isolate USA-WA1/2020 at approximately 3x LoD. 3 bacteria and 18 viruses were tested.
All negative samples gave negative results with the potentially cross-reactive common organisms tested showing no cross-reactivity with SARS-CoV-2 COVID-19 Antigen assay. All samples with SARS-CoV-2 strain tested positive showing no microbial interference at the concentrations of the potentially interfering common organisms tested.

Analytical Specificity Cross Reactivity (Exclusivity) and Microbial Interference:
The potential cross-reactivity (exclusivity) of a panel of common organisms was evaluated with SARS-CoV-2 negative samples using the SARS-CoV-2 COVID-19 Antigen test. Potential microbial interference was evaluated with samples containing heat-inactivated SARS-CoV-2 isolate USA-WA1/2020 at approximately 3x LoD. 3 bacteria and 18 viruses were tested.
All negative samples gave negative results with the potentially cross-reactive common organisms tested showing no cross-reactivity with SARS-CoV-2 COVID-19 Antigen assay. All samples with SARS-CoV-2 strain tested positive showing no microbial interference at the concentrations of the potentially interfering common organisms tested.

Potential Cross Reaction

Negative Specimen

Spiked with SARS-CoV-2

Adenovirus Type 3

Negative

Positive

Influenza A H1N1

Negative

Positive

Influenza A H3

Negative

Positive

Influenza A 2009 H1N1

Negative

Positive

Influenza B

Negative

Positive

Metapneumovirus 8

Negative

Positive

Respiratory Syncytial Virus A

Negative

Positive

Rhinovirus 1A

Negative

Positive

Parainfluenza virus type 1

Negative

Positive

Parainfluenza virus type 2

Negative

Positive

Potential Cross Reaction

Negative Specimen

Spiked with SARS-CoV-2

Parainfluenza virus type 3

Negative

Positive

Parainfluenza virus type 4

Negative

Positive

Coronavirus NL63

Negative

Positive

Coronavirus 229E

Negative

Positive

Coronavirus OC43

Negative

Positive

Coronavirus HKU-1

Negative

Positive

B. pertussis

Negative

Positive

C. pneumonia

Negative

Positive

M pneumonia

Negative

Positive

Interfering substances
To assess substances with the potential to interfere with the performance of the SARS-CoV-2 COVID-19 Antigen, positive and negative samples were tested with the addition of potential interfering substances. The SARS-CoV-2 target concentration in the positive samples was approximately 3x LoD. All samples tested produced expected results, demonstrating that the SARS-CoV-2 COVID-19 Antigen test performance was not affected by any of the 22 potentially interfering substances listed in the table below at the concentrations tested.

Interfering substance

Final Concentration

Negative Specimen

Spiked with SARS-CoV-2

Benzocaine

150 mg/dL

Neg.

Pos.

Blood

5%

Neg.

Pos.

Mucin

5 mg/mL

Neg.

Pos.

NasoGEL

5%

Neg.

Pos.

Phenylephrine

15%

Neg.

Pos.

Oxymetaoline

15%

Neg.

Pos.

NasalCrom

15%

Neg.

Pos.

Zicam Cold Remedy

5%

Neg.

Pos.

Alkalol (homeopathic)

10%

Neg.

Pos.

Chloraseptic

15%

Neg.

Pos.

Tobramycin

3.3 mg/dL

Neg.

Pos.

Interfering substance

Final Concentration

Negative Specimen

Spiked with SARS-CoV-2

Mupirocin

0.15 mg/dL

Neg.

Pos.

Fluticasone

0.000126 ug/dL

Neg.

Pos.

Tamiflu

500 mg/dL

Neg.

Pos.

Budenoside

0.00063 mg/dL

Neg.

Pos.

Biotin

0.35 mg/dL

Neg.

Pos.

Methanol

150 mg/dL

Neg.

Pos.

Acetylsalicyclic Acid

3 mg/dL

Neg.

Pos.

Diphenhydramine

0.0774 mg/dL

Neg.

Pos.

Dextomethorphan

0.00156 mg/dL

Neg.

Pos.

Dexamethasone

1.2 mg/dL

Neg.

Pos.

Mucinex

5%

Neg.

Pos.

Cross Reactivity with SARS-CoV-1
The SARS-CoV-2 COVID-19 Antigen was tested with heat inactivated SARS-CoV-1 (strain 2003-00592/lot 324780) by Zeptometrix. The Test was successful since E-gene exhibited a positive value of 26.76 CT value with both RdRP & N Gene exhibiting a Ct value.

The SARS-CoV-2 COVID-19 Antigen exhibits NO cross reactivity with SARS-C0V-1.

Specimen Stability (Refrigerated & Room Temperature)
The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV- 2 Antigen Test was evaluated with SARS-CoV-2 negative and positive specimens. The stability at room temperature was evaluated by placing the samples in a dry tube and stored at 24 – 27o C (room temperature) or 4o C refrigerated for 0, 1, 3 and 7 days.

Specimen Stability (Refrigerated & Room Temperature)
The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV- 2 Antigen Test was evaluated with SARS-CoV-2 negative and positive specimens. The stability at room temperature was evaluated by placing the samples in a dry tube and stored at 24 – 27o C (room temperature) or 4o C refrigerated for 0, 1, 3 and 7 days.

These studies demonstrated that storage for up to 7 days at either temperature did not have an effect on the stability of the SARS-CoV-2 samples when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen Te s t and the comparator PCR essay.

Stored at 24-270 C

Positive Samples

Negative Samples

Days

Positive

Negative

Positive

Negative

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

3

10

0

0

5

Stored at 24-270 C

Positive Samples

Negative Samples

Days

Positive

Negative

Positive

Negative

4

10

0

0

5

5

10

0

0

5

6

10

0

0

5

7

10

0

0

5

Comparator PCR

Stored at 24-270 C

Positive Samples

Negative Samples

Days

Positive

Negative

Positive

Negative

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

3

10

0

0

5

Stored at 24-270 C

Positive Samples

Negative Samples

Days

Positive

Negative

Positive

Negative

4

10

0

0

5

5

10

0

0

5

6

10

0

0

5

7

10

0

0

5

Specimen Stability (Freeze/Thaw)
The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen Test was evaluated with SARS-CoV-2 negative and positive specimens. The stability during freeze/thaw cycles was evaluated by placing the samples in a dry tube and stored at -16oC (freeze) then thawed five cycles to 24 – 27o C (room temperature).
These studies demonstrated that extreme volatility freeze/thaw cycles did not have an effect on the stability of the SARS-CoV-2 Antigen Test when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen and the comparator PCR assay.

Specimen Stability (Freeze/Thaw)
The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen Test was evaluated with SARS-CoV-2 negative and positive specimens. The stability during freeze/thaw cycles was evaluated by placing the samples in a dry tube and stored at -16oC (freeze) then thawed five cycles to 24 – 27o C (room temperature).
These studies demonstrated that extreme volatility freeze/thaw cycles did not have an effect on the stability of the SARS-CoV-2 Antigen Test when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen and the comparator PCR assay.

-16 0 C to 27 0 C

Positive Samples

Negative Samples

volatility

Cycles

Pos.

Neg.

Pos.

Neg.

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

-16 0 C to 27 0 C

Positive Samples

Negative Samples

volatility

Cycles

Pos.

Neg.

Pos.

Neg.

3

10

0

0

5

4

10

0

0

5

5

10

0

0

5

Comparator PCR

-16 0 C to 27 0 C

Positive Samples

Negative Samples

volatility

Cycles

Pos.

Neg.

Pos.

Neg.

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

-16 0 C to 27 0 C

Positive Samples

Negative Samples

volatility

Cycles

Pos.

Neg.

Pos.

Neg.

3

10

0

0

5

4

10

0

0

5

5

10

0

0

5

Specimen Stability (Extreme Heat) to 40oC and back to RT

The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV- 2 Antigen was evaluated with SARS-CoV-2 negative and positive specimens. The stability during consistent extreme heat was evaluated by placing the samples in a 40 oC incubator and allowed to acclimate and then removed from incubator and acclimated to Room Temperature for 30 minutes. This procedure was repeated 5 cycles.

These studies demonstrated that extreme heat did not have an effect on the stability of the SARS-CoV-2 when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen and the comparator PCR assay.

These studies demonstrated that extreme heat did not have an effect on the stability of the SARS-CoV-2 when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen and the comparator PCR assay.

Room Temp to

Extreme Heat at 400C

Positive Samples

Negative Samples

Cycles

Pos.

Neg.

Pos.

Neg.

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

Room Temp to

Extreme Heat at 400C

Positive Samples

Negative Samples

Cycles

Pos.

Neg.

Pos.

Neg.

3

10

0

0

5

4

10

0

0

5

5

10

0

0

5

Comparator PCR

Room Temp to

Extreme Heat at 400C

Positive Samples

Negative Samples

Cycles

Pos.

Neg.

Pos.

Neg.

0

10

0

0

5

1

10

0

0

5

2

10

0

0

5

Room Temp to

Extreme Heat at 400C

Positive Samples

Negative Samples

Cycles

Pos.

Neg.

Pos.

Neg.

3

10

0

0

5

4

10

0

0

5

5

10

0

0

5

Specimen Stability (Extreme Heat) sustained long term at 40oC
A 2-year long term long term heat study at 40oC is currently underway. The specimen stability of nasopharyngeal swab in extraction buffer for Rapid Diagnostic Test for the Detection of SARS-CoV- 2 Antigen was evaluated with SARS-CoV-2 negative and positive specimens. The stability during consistent extreme heat was evaluated by placing the samples in a 40oC incubator long term. A subset of the kits and reagent are tested at the time points noted below.

These studies demonstrated that extreme heat of 40oC sustained for One (1) Month does not have an effect on the stability of the SARS-CoV-2 when tested against the Rapid Diagnostic Test for the Detection of SARS-CoV-2 Antigen Test Kit and the comparator PCR assay.

Stored at 40oC

Positive Samples

Negative Samples

Weeks

Pos.

Neg.

Pos/Neg

Neg.

1

10

0

0/5

5

2

10

0

0/5

5

4

10

0

0/5

5

12

pending

pending

pending

pending

Stored at 40oC

Positive Samples

Negative Samples

Weeks

Pos.

Neg.

Pos/Neg

Neg.

24

pending

pending

pending

pending

52

pending

pending

pending

pending

104

pending

pending

pending

pending