GLOW+ COVID-19 Antigen Rapid Test Instructions for Use

GLOW+ COVID-19 Antigen Rapid Test

Instructions for Use

Version: 1.0                     Release Date: 2020/04/03
Doc #: 20-03-100            REF 300100

20
Panacyte Scientific LLC
341 Science Park Rd. Ste 210, State College, PA 16802
+1-814-441-5277
support@panacyte.com

www.panacyte.com

Attention: Please Read this Instruction for Use before conducting test.

INTENDED USE

Panacyte Glow+ COVID-19 Antigen Rapid Test is a fluorescent-enhanced lateral flow immunoassay intended for the presumptive qualitative detection of nucleocapsid protein from the 2019-nCoV in nasopharyngeal swab, oropharyngeal swab and sputa from individuals meeting CDC 2019-nCoV clinical criteria (e.g., clinical signs and symptoms associated with 2019-nCoV infection) in conjunction with CDC 2019-nCoV epidemiological criteria (e.g., history of residence in or travel to a geographic region with active 2019-nCoV transmission at the time of travel, or other epidemiologic criteria for which 2019-nCoV testing may be indicated). Testing is limited to laboratories – certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42
U.S.C. §263a, to perform high complexity tests, or by similarly qualified non-U.S. laboratories.
Results are for the detection and identification of 2019-nCoV antigen(s). The 2019- nCoV nucleocapsid protein is generally detectable in nasopharyngeal swab, oropharyngeal swab or sputa during the acute phase of infection. Positive results are
indicative of active infection. Laboratories within the United States and its territories are required to report all positive results to the appropriate public health authorities.
Negative results do not preclude 2019-nCoV infection and should not be used as the sole basis for patient management decisions. Negative results must be combined with clinical observations, patient history, and epidemiological information.

STORAGE

• Panacyte Glow+ COVID-19 Antigen Rapid Test can be stored at 4°C-30°C for up to 6 months
• Avoid direct sunlight to the exposed Panacyte Glow+ COVID-19 Antigen Rapid Test device

REAGENTS

Components below are included in the Glow+ COVID-19 Antigen Rapid Test kit

The lysis buffer contains PH7.4 Tris-HCL, NaCl, NP40, Sodium Deoxycholate, EDTA.

Needed but not provided

1. Timer

Positive Control

Positive control samples can be separately purchased, with REF 300101.

PRINCIPLE OF THE TEST

Panacyte Glow+ COVID-19 Antigen Rapid Test is a lateral flow immunoassay. Polyclonal antibody specific to the nucleocapsid protein of the SARS-CoV-2 virus was covalently labeled with 300nm-diameter Time Resolved Fluorescent Microspheres (TRFM). The TRFM is filled with rare earth ions that has much higher fluorescence upon excitation. Usage of TRFM increases signal strength for low-level viral antigen, hence increases the sensitivity of the test. Fig 1 illustrates the Glow+ COVID-19 Antigen Rapid Test Strip inside the device cartridge. In the direction of the sample flow, the strip contains a sample pad, antibody-tracer pad, cellulous nitrate (CN) membrane, and last absorption pad. Two antibody lines, the T line (test line) and the C line (control line), were immobilized on the NC membrane by spray printing. The antibody-tracer pad contains TRFM-conjugated polyclonal IgG that is antiN protein (Ab1-TRFM complex). The T line was spray painted with the same polyclonal IgG (Ab2). The C line was spray painted with anti-IgG antibody. After sample was introduced, assuming it contains N protein, N proteins will be captured by the TRFM-Ab1 and form NP-Ab1-TRFM complex. Sample fluid carries the NP-Ab1- TRFM complex into the NC membrane. When flowing through the T line, the immobilized Ab2 will capture the NP-Ab1-TREM, forming Ab2-NP-Ab1-TRFM. Unbounded Ab1-TRFM and surplus NP-Ab1-TRFM will continue flow to the C line, where the anti-IgG antibody will capture Ab1-TRFM and NP-Ab1-TRFM. The two capturing processes will continue until all sample fluid has passed and absorbed by the absorption pad.

The entire lateral flow process will take 12 mins to complete.

SAMPLE AND SAMPLE COLLECTION

Glow+ COVID-19 Antigen Rapid Test system for the SARS-CoV-2 can accept the following samples：

• Nasopharyngeal swab (recommended for early stage patients [Ref. 1])
• Oropharyngeal swab

Special notes for swab sample stored in storage/transport media

Swab sample stored in storage/transport media will decrease the test sensitivity, due to the dilution of the virus concentration by larger volume of the media. It is thus not recommended to be used for this test. If a media- stored swab sample has to be used, it is recommended taking the following steps:

1. Allow the vial of swab-containing media to return to room temperature
2. Manually mix the vial for one minute
3. Use pipette to aspirate 150-microliter of mixed media into the Glow+ COVID19 Antigen Rapid Test Lysis Buffer bottle
4. Follow from Step 6 of the protocol
5. Remember, this can cause false negative

Following Specimen can NOT be used:

• NP Swab specimen stored in RNA extraction can NOT be used, due to the presence of proteinases. May lead to False Negative.
• Nasopharyngeal swab taken improperly or by untrained professionals may lead to false negative results.
• Nasopharyngeal swab taken with excessive blood

Tips for Better Nasopharyngeal Sample Collection

1. Clear nose with tissue before sample collection
2. Title head back 30 degree off vertical, and push the swab horizontally in
3. Push the NP swab all the way to the back (3 to 4 inches). One way to estimate how deep it should go in is by measuring the straight distance from tip of nose to the bottom tip of the earlobe, using a stiff ruler, then minus half inch.

Image was taken from YouTube, published by The Joint Commission on Oct. 15, 2012. Permission Pending

Fig. 1 Collect Nasopharyngeal swab specimen, making sure the swab penetrates deeply to the end

PROTOCOL (NASOPHARYNGEAL SWAB)

1. Collect patient’s nasopharyngeal or pharyngeal samples using the included sterile swab
2. [Fig. 2, Step 1] Twist open the colored top of the drip bottle containing the lysis buffer. Do not remove the clear top cap.
3. [Fig. 2, Step 1] Insert the swab to the bottle.
4. [Fig. 2, Step 2] Snap the swab at the notch (3 inches from the tip) and leave the front half in the bottle.
5. [Fig. 2, Step 3] Use previously removed bottle top to push in the front half of the swab, twist it all the way to close the bottle.
6. Gently swirl the bottle for 2 minutes for the lysis buffer to work.
7. [Fig. 3] Check the color of the lysis buffer. It should be clear or slightly cloudy (double check marks). It should not appear to be red, which may be caused by nosebleed. If recollecting NP swab is not possible, slightly yellow-red buffer can be used (one check mark).
8. Remove the test device from the pouch and leave it on a flat surface.
9. [Fig. 2, Step 4] Remove the clear top cap.
10. [Fig. 2, Step 4] Carefully invert the bottle, above the Sample Well of the device。
11. [Fig. 2, Step 4] Gently squeeze the body of the drip bottle, to allow 5 drips of the lysis buffer free fall into the sample well.
12. Wait 15 minutes for the lateral flow process to complete.
13. [Fig. 4] Illuminate the observation window with 365nm UV light. Identify thebrighter C line, and then the T line that is 1/8”  closer to the sample well, using [Fig. 5] as a guidance.
14. If test is negative, wait another 5 minutes and re-read the results.

Fig 2. To Use the Glow+ COVID-19 Antigen Rapid Test

(May not resemble the final product)

Fig 3. Inspect the lysis buffer before adding to the test device

LIMIT OF DETECTION (LoD)

20ng Nucleocapsid Protein per test device
Or (equivalent to)

100ng Nucleocapsid Protein per bottle of Lysis Buffer

RESULT INTERPRETATION

Step 1. Identify the Control Line (labeled C in [Fig. 4]). If Control Line can’t be found, the test is NOT valid. If Control Line presents, move on to Step 2.
Step 2. Find the Test Line, as indicated by the arrows in [Fig. 4] and make decision of “Negative” or “Positive” using reference images provided in [Fig. 5].
If Control Line is missing
If the control line cannot be found, please following these steps

1. Please check if the 365nm wavelength UV flashlight has insufficient power
2. Please run a negative control test
3. If negative control test fails, please follow the steps outlined in the QUALITY CONTROL section.
4. If negative control test is successful, repeat the test with a freshly collected swab specimen.

Special notes for result interpretation

1. A negative result can only be determined after step 14 of the protocol and if there is absolutely no sign of Test Line. Please reference all images provided in the Positive section in [Fig.5].
2. Streaming ([Fig. 5] Panel A.1. and Panel B.1.) should be reduced after allowing longer time as in step 14. You may allow up to 20 minutes.
3. A wake positive (dim T line) should be treated as positive. Please pay special attention to images in [Fig. 5] Panel B.3.
4. With a positive specimen, the T line may develop before the 15 min lateral flow time, as in step 12 of the protocol. When T line is observed before 15 min, the test can be determined as positive at this time and thus completed.

QUALITY CONTROL

Positive Control

Use Positive Control (purchased separately) and a vial of lysis buffer, Add lysis buffer to the Positive Control tube, swirl until dry powder fully dissolve. Using transfer pipette to move all solution back into the lysis buffer bottle. Follow protocol step 7 thru 14. Both C line and T line should be visible under 365nm wavelength UV illumination.

Negative Control

Use provided sterile swab, without collecting any samples, follow protocol step 2 thru 14. Only C line should be visible under 365nm wavelength UV illumination.

Recommended Action for Control Failure

In general, any failure in either negative control test or positive control test, should prevent subsequent usage of the test kits with the same lot number within the same facility. Failed control test can be in one of the 4 modes below.

1. Positive control test with C line and no T line, including questionable T line: (step 1) check the expiration date of the control sample. If not expired, (step 2) check if the storage condition has been violated. If not, (step 3) The test strip has failed. The same box of test kit should not be used.

2. Positive control test with no C line and no T line: (step 1) Make sure lateral flow has taken place (the strip inside the observation window appears wet). If not, repeat the test and make sure 4 to 5 drops of sample is added to the sample well. If still no lateral flow, the same box of test kit should not be
   used. (step 2) The test strip has failed. The same box of test kit should not be used.

3. Negative control test with a C line and a T line: The likely cause is bad lysis buffer. The same box of test kit should not be used.

4. Negative control test with no C line and no T line: (step 1) Make sure lateral flow has taken place (the strip inside the observation window appears wet). If not, repeat the test and make sure 4 to 5 drops of sample is added to the sample well. If still no lateral flow, the same box of test kit should not be used. (step 2) The test strip has failed. The same box of test kit should not be used.

INTERFERENCE SUBSTANCES

1. Swab media containing proteases will abolish the formation of T line.
2. More than 5 microliters of blood in the collected sample willabolish the formation of T line.
3. Fluticasone Propionate (allergy nasal spray, commonly known as Flonase) may weaken the test lines
4. Allergy eyedrops do not interfere with the assay
5. Saliva and tear do not interfere with the assay

WARNINGS

1. During the processes of sample collection, please follow your local SOP and guidance. Personal protective equipment (PPE) must be used. Avoid crosscontamination.
2. Upon completion of the lateral flow, T line and C line are not visible without UV excitation.
3. When 365nm UV flashlight has insufficient battery, the test lines may not be visible. Please always use a fully charged flashlight or use with a new battery.
4. Avoid directly shine the UV light into eyes.
5. Negative or positive determination serves as a supplemental test to the nucleicacid test. Clinical diagnosis should be reached in conjunction with symptoms and according to local diagnostic guideline for COVID-19.

DECLARATIONS AND LIMITATIONS

• Currently, the detection of viral protein can only be used as a supplemental test, not to confirm or diagnose.
• If sample contains high concentration of virus, T line may beabsent (false negative). At such condition, COVID-19 symptoms should be present and be considered when reaching a diagnostic determination.
• Antihistamine nasal spray, such as Flonase, may decrease the signal strength of test lines
• Excessive nosebleed will interfere with the assay. Do NOT take NP Swab from
• The comparative study between SARS-CoV-2 and other coronaviruses, including SARS, MERS and common cold, has not been conducted on the Glow+ COVID-19 Antigen Rapid Test.
• Please dispose used NP swab, lysis buffer drip bottle and test device in biohazard waste container.

Fig 4. Panacyte Glow+ COVID-19 Antigen Rapid Test, Negative and Positive

Reference Images

A. Negative

Panel A.1. and Panel A.2. are both negative results
Panel A.1. Negative Test

Panel A.2. Negative Test with Streaming

B. Positive

Panel B.1., B.2., B.3., B.4. are all Positive Results
Panel B.1. Positive Test

Panel B.2. Positive Test with Streaming

Panel B.3. Weak Positive Test

Panel B.4. Positive Test – Analyte Quantity High to Low

Fig 5. Reference images of negative, weak positive and positive determinations

REFERENCE

1. Clinical presentation and virological assessment of hospitalized cases of coronavirus disease 2019 in a travelassociated transmission cluster, R. Woelfel, et.al MedRxiv, posted online on March 08, 2020
2. The severe acute respiratory syndrome coronavirus 3a is a novel structural protein, S. Shen, Y-J Tian, Biochem and Biophy Res Comm, 2005, 330[1], pp286-292
3. Influenza Specimen Collection, FDA, flu-specimen-collection-poster [pdf]
4. A Novel Coronavirus from Patients with Pneumonia in China, Na Zhu, et.al, 2019, N Engl J Med 2020; 382:727-733

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