COVID-19 Antigen Detection Kit (Latex Lateral Flow Assay)

Product stability research data

Shenzhen Aivd Biotechnology Co., LTD

Directory
Directory …………………………………………………………………………………………………2
1.Research data on the stability of COVID-19 antigen detection kit (latex method) products… 3
1). Basis for determination of stability research method …………………………………………… 3
2). Stability study Scheme …………………………………………………………………………… …3
3）The specific method and process of stability study ………………………………………………4
1. Stability assessment at storage temperature……………………………………………………… 4
1.1 Experimental Materials ………………………………………………………………………………4
1.2 Experimental Method …………………………………………………………………………………4
1.3 Test results should meet the standard ………………………………………………………………4
2. 37 ℃ accelerated thermal stability test ………………………………………………………………4
2.1 Experimental materials ………………………………………………………………………………4
2.2 Experimental Method……………………………………………………………………………… …4
2.3 Test results should meet the standard ………………………………………………………………5
3. Simulated vehicle-mounted road transport stability test…………………………………………… 5
3.1 Background…………………………………………………………………………………………… 5
3.2 Experimental material ………………………………………………………………………………5
3.3 Experimental Method …………………………………………………………………………………5
4. Stability test after opening …………………………………………………………………………5
4.1 Experimental materials ……………………………………………………………………………… 6
4.2 Experimental Method ……………………………………………………………………………… 6
Quality standards to be achieved by stability testing ………………………………………………… 6
Handling method when the stability assessment is inconsistent …………………………………… 6
Stability test and data statistics ……………………………………………………………………… 7
1. Stability assessment at storage temperature (2 ~ 30) ℃ ………………………………………… 7
2. Thermal stability test for 37℃ acceleration …………………………………………………………11
3. Simulated vehicle-mounted road transport stability test ………………………………………… 13
4. Stability test after opening during the experiment …………………………………………………14
Analysis of stability assessment results ……………………………………………………………15
Summary and conclusion of stability assessment ……………………………………………………15

1．Data from a study on the stability of the product of the COVID-19 antigen detection kit (latex method)
1) the basis for determining the stability research method
El23640:2002 Stability Testing of in Vitro Diagnostic Reagents.
2). Stability study scheme
The stability study of kit is mainly to investigate the quality change of kit with the increase of storage time
under the specified storage conditions, and determine the validity period of kit under the specified storage
conditions according to the obtained data.
Stability study test design:
1.1 The storage condition of this kit is sealed and stored at (2 ~ 30) ℃ for 12 months. Therefore, the
quality change of the product with the increase of storage time to the 15th month under this storage
condition is designed.
1.2. Accelerated thermal stability test at 37 ℃ : The kit was stored in a constant temperature environment
of 37℃ for 15 days, and tested in stages during the storage;  The kit should be kept at 37℃ for more than
10 days.
1.3. Vehicle transport process simulation stability test: This reagent needs to be sent to domestic provinces
and cities by vehicle, the longest vehicle time is 5 ~ 7 days. Therefore, we placed the kit on the company's
medium bus for 10 days, and investigated the product performance stability of the kit simulating
environmental changes such as temperature and vibration during road transportation. The kits were then
stored at (2 ~ 8) ℃ and (8 ~ 30) ℃ until the sixth month and the 12th month of the validity of the kits. The
stability of the kits after vehicle transportation was investigated.
1.4. After opening the experiment process stability test: place the kit product after opening experiment in
kit regulation of temperature and humidity operating environment (temperature 20 ℃ to 25 ℃, humidity
35% ~ 65%) in the next period of time, then test, inspect the kit experimental process after opening is
placed under the temperature and humidity conditions of a specific test operation stability.
1.5. Stability test at storage temperature after opening: the kit uses a test card to assemble aluminum foil
bag and seal the package separately. There is no case of repeated use after opening.  Therefore, this test
scheme does not involve the stability test of the product after repeated use at the storage temperature after
opening.

3）. Specific methods and processes of stability research

1. Stability assessment at storage temperature
1.1 Experimental materials
The kits for stability study are batch numbers 20200301,20200302, 20200303, and the specifications are 1
PCS/bag, 25 PCS/box.
Internal reference products: 202003 batches
1.2 Experimental methods
Six batches of finished COVID-19 antigen detection kits (latex method) were extracted from each batch of
108 people, stored in cold storage environment (2-8) ℃ and workshop environment (8-30) ℃ for 15
months respectively, and tested once at 0, 1, 3, 6, 10 and 15 months respectively. Internal reference
samples of the enterprise were used as assessment samples, and the test items were carried out according to
the stability test should meet the quality standard of finished products of the project. The change of kit
quality with the increase of storage time was investigated. Finally, the detection results are summarized
statistically.
1.3 Test results should meet the standards
The kit should be stored at storage temperature (2 ~ 30) ℃ for more than 12 months, that is, the quality of
the kit must still meet the quality standard of finished product verification after 12 months of storage under
this condition.

2. 37 ℃ acceleration thermal stability test

2.1 Experimental materials
COVID-19 antigen detection kit (latex method), batch number: 20200301,20200302, specifications: 1
PCS/bag, 25 PCS/box 20200303.
Internal reference products: 202003 batches
2.2 Experimental methods
54 samples from each batch of the finished COVID-19 antigen detection kit (latex method) were extracted
and stored in a constant temperature environment of 37 ℃ for 15 days. Tests were performed on the first,
second, third, sixth, tenth and fifteenth days respectively. Internal reference samples of the enterprise were
used as assessment samples, and the test items were carried out according to the stability test should meet
the quality standard of finished products of the project, and the change of kit quality with the increase of
storage time was investigated. Finally, the detection results are summarized statistically.
2.3 Standards that test results should meet

The kit should be kept at a constant temperature of 37 ℃ for more than 10 days, that is, after 15 days of
storage under this condition, the quality of the kit must still reach (iv). Quality standards to be achieved by
stability testing).
3. Simulated vehicle road transport stability test
3.1 Related Background
Because the developed product is sent to domestic provinces and cities by road transportation, the longest
road transportation time involved is 5 ~ 7 days. Therefore, we designed the following simulation
conditions: a batch of reagent finished products were placed on the company's shuttle bus for 10 days, and
the main driving distance of the company's shuttle bus was: national road, highway, township road,
etc. During the experiment, the temperature and humidity conditions and the mileage of the minibus were
recorded every day. The product performance stability of the kit was investigated by simulating the
environmental changes of temperature and vibration during road transportation. The kits were then stored
at (2 ~ 8) ℃ and (8 ~ 30) ℃ until the sixth month and the 12th month of the validity of the kits. The
stability of the kits after vehicle transportation was investigated.
3.2 Experimental materials
COVID-19 antigen detection kit (latex method), lot no. 20200301,20200304, size: 1 PCS/bag, 25 PCS/box.
Internal reference products: 202003 batches
3.3 Experimental methods
A total of 2,020,301 batches of COVID-19 antigen detection kit detection cards were randomly selected
from the finished products and placed on the company's shuttle bus to inspect the stability of the simulated
transportation process of the kit. After 10 days of simulated road transportation, the kits were stored at (2-
8) ℃ and (8-30) ℃ until the 6th and 12th month of reagent validity period.  Internal reference samples of
the enterprise were used as assessment samples, and the test items were carried out according to the
stability test should meet the quality standard of finished products of the project, and the change of kit
quality with the increase of storage time was investigated. Finally, the detection results are summarized
statistically.

4. Stability test after opening

4.1 Experimental Materials
COVID-19 Antigen Detection Kits (latex method), lot numbers 20200301 and 20200303, specifications: 1
PCS/bag, 25 PCS/box.
Internal reference products: 202003 batches.

4.2 Experimental Methods

A total of 2020,301 batches of finished COVID-19 antigen detection kits (30 persons) were randomly
selected and placed in the operating environment specified by the kits (temperature 20-25 ℃, humidity 35-
65%). After the reagent detection cards were balanced for 5-10 minutes, the sealed aluminum foil bag of
the detection cards was torn open and exposed to the temperature and humidity environment. The test was
carried out at 0, 5, 15, 30 and 60 minutes after tearing the package. The internal reference samples of the
enterprise were used as assessment samples, and the test items were carried out according to the finished
product quality standards of the project for stability testing. The change of the kit with time was
investigated when the kit was placed in the specified operating environment (temperature 20-25 ℃,
humidity 35-65%) after opening in the experimental process. Finally, the detection results are summarized
statistically.

4）. Quality standards to be met by stability testing

1. Liquid transit time: use the kit of the same batch to conduct quantitative test on the reference product in
the enterprise, and start timing with liquid dropping into the sample well. The time required for liquid to
pass through the observation window should be less than 10 minutes.
2. Accuracy: Use enterprise internal control accuracy reference products (L2,L2,L3) to test, the test results
should be within the following range:
N1 (negative) : using the enterprise internal control accuracy reference NI test, in 15 minutes, T band does
not display, C band color should be close to the control;
L3 (low) : by using the enterprise internal control accuracy reference L3 test, at 15 minutes, T band and C
band color should be similar to the control;
L1 (strong) : detected by L1, the color display of T band and C band should be close to the control at 15
minutes;

5）. Handling method when the stability assessment does not meet the requirements
The inspection process shall be operated strictly in accordance with the instructions. If the test results do
not conform to the provisions of internal control reference products, the reasons shall be found first. After
finding out the reasons, the non-conforming samples shall be retested for three times, and the judgment
shall be made according to the retest results.
Strictly follow the temperature and humidity environment for reagent operation, the temperature is 20-25
℃, and the relative humidity is controlled between 35-65%.
6）. Stability assessment test and data statistics
1. Stability assessment under storage temperature (2 ~ 30) ℃

1.1Liquid migration time

 lot TEST 1 TEST2 TEST3

20200301 1'38" 1'49" 1'34"

20200302 1'37" 1'51" 1'35"

20200303 1'48" 1'43" 1'29"

Eligibility criteria：<10min result meets requirements

1.2 Stability results of the kit at storage temperature (2-30) ℃

Table 1 Stability results of kit no. 20200301 at storage temperature (2 ~ 30 ℃)
Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
0months
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
1months
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
3months Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
8～30℃ Quality N1 （ negative L3（ positive L1（positive Yes
controls ） ） ）
Parallel test - + +++
three times - + +++
 - + +++
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ） Yes
2～8℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
6months
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
10months
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
15months
Quality N1 （ negative L3（ positive L1（positive
controls ） ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
(Note: Test result "+++" means strong positive; The test result of "++" means medium positive; The test
result of "+" means weak positive; A test result with a "-" means negative.)

Table 2 Stability results of kit batch No. 20200302 at storage temperature (2-30 ℃)
Storage storage Yes or no meet
results
time conditions requirements
0months 2～8℃ Quality N1 （ negative L3 （ positive Yes
L1（positive）
controls ） ）
Parallel test - + +++
three times - + +++
- + +++
 Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
1months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
3months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
6months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
10months Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
2～8℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
8～30℃ Quality N1 （ negative L3 （ positive L1（positive） Yes
controls ） ）
 - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
15months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
(Note: Test result "+++" means strong positive; The test result of "++" means medium positive; The test
result of "+" means weak positive; A test result with a "-" means negative.)

Table 3 Stability results of batch no. 20200303 kit at storage temperature (2-30 ℃)
Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
0months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
1months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
3months 2～8℃ Quality N1 （ negative L3 （ positive L1（positive） Yes
controls ） ）
 - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
6months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
2～8℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
10months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
8～30℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
Yes
2～8℃ - + +++
Parallel test
- + +++
three times
- + +++
15months
Quality N1 （ negative L3 （ positive
L1（positive）
controls ） ）
8～30℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
(Note: Test result "+++" means strong positive; The test result of "++" means medium positive; The test
result of "+" means weak positive; A test result with a "-" means negative.)
 2. Thermal stability test for acceleration at 37℃
Table 4 Results of accelerated thermal stability of kit 20200301 at 37℃
Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 1 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 2 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 3 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Yes
Day 6 37℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Yes
Day 10 37℃ - + +++
Parallel test
- + +++
three times
- + +++
Day 15 37℃ Quality N1 （ negative L3 （ positive L1（positive Yes
controls ） ） ）
Parallel test - + +++
three times - + +++
- + +++

(Note: Test result "+++" means strong positive; The test result of "++" means medium positive; The test
result of "+" means weak positive; A test result with a "-" means negative.)

Table 5 Results of accelerated thermal stability of kit 20200301 at 37℃

Storage storage Yes or no meet
results
time conditions requirements
 Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 1 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 2 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 3 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Yes
Day 6 37℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ） Yes
Day 10 37℃
Parallel test - + +++
three times - + +++
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 15 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++

Table 6 Results of accelerated thermal stability of kit 20200301 at 37℃

Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 1 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Day 2 37℃ Quality N1 （ negative L3 （ positive L1（positive Yes
 controls ） ） ）
- + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 3 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Yes
Day 6 37℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Yes
Day 10 37℃ - + +++
Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Day 15 37℃ - + +++ Yes
Parallel test
- + +++
three times
- + +++

3. Simulated vehicle road transport stability test

Table 7 Stability results of kit simulation for vehicle-mounted road transport, Batch No. 20200301 
Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3 （ positive L1（positive
controls ） ） ）
Ten days
8～30℃ - + +++ Yes
in the car Parallel test
- + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
2～ After 10 controls ） ） ）
8℃6mo days in the - + +++ Yes
Parallel test
nths car - + +++
three times
- + +++
2～8℃ After 10 Quality N1 （ negative L3 （ positive L1（positive Yes
12month days in controls ） ） ）
 - + +++
Parallel test
s the car - + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
After 10 controls ） ） ）
8～30℃
days in - + +++ Yes
6months Parallel test
the car - + +++
three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
After 10
8～30℃ controls ） ） ）
days in Yes
months Parallel test - + +++
the car
three times - + +++
- + +++

4. Stability test after opening during the experiment

Table 8 Stability results of kit after opening, Lot no. 20200301
Storage storage Yes or no meet
results
time conditions requirements
Quality N1 （ negative L3 （ positive L1（positive
Temperature 20
controls ） ） ）
~ 25℃,
0min - + +++ 符合
humidity 35 ~ Parallel test
- + +++
65%) three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
Temperature 20
controls ） ） ）
~ 25℃,
5min - + +++ 符合
humidity 35 ~ Parallel test
- + +++
65%) three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
Temperature 20
controls ） ） ）
~ 25℃,
15min - + +++ 符合
humidity 35 ~ Parallel test
- + +++
65%) three times
- + +++
Quality N1 （ negative L3 （ positive L1（positive
Temperature 20
controls ） ） ）
~ 25℃,
30min - + +++ 符合
humidity 35 ~ Parallel test
- + +++
65%) three times
- + +++
60min Temperature 20 Quality N1 （ negative L3 （ positive L1（positive 符合
~ 25℃, controls ） ） ）
humidity 35 ~ Parallel test - + +++
 - + +++
three times
65%) - + +++
(Note: Test result "+++" means strong positive; The test result of "++" means medium positive; The test
result of "+" means weak positive; A test result with a "-" means negative.)

7）Analysis of stability assessment results

1. When stored at 2-30 ℃, the kit was stored for 15 months and all indicators were in line with the quality
standards.
2. The kit was tested at 37℃ for 15 days, and all indicators were in line with the quality standards.
3. Vehicle road transportation process simulation stability test. The kits simulated road transportation for 10
days were stored at (2 ~ 8)℃ and (8 ~ 30)℃ until the 6th and 12th month of the validity of the kits for
testing.
4. After the finished kit was unsealed, it was placed in the operating environment (20-25 ℃, 35-65%
humidity) specified by the reagent in the room for one hour, and all indicators were in line with the quality
standards.

8）Summary and conclusion of stability assessment

The comprehensive assessment results showed that the stability of the reagent stored at (2 ~ 30 ℃), the
accelerated thermal stability of the reagent at 37 ℃, the simulation stability of the reagent during
transportation, and the unsealing stability of the reagent during experiment. The stability of our COVID-19
antigen test kit (latex method) fully meets the expected quality standards, That is, storage at 2-30 ℃ for 15
months, thermal acceleration damage at 37℃ for 15 days, and 10 days after the simulated vehicle
transportation by road, continue to store at (2-8) ℃ and (8-30) ℃ until the 12th month of the validity of the
kit, and store in the specified operating environment conditions of the reagent after opening (20-25
℃, Humidity 35 ~ 65%) reagent within one hour, all the test indicators are in line with the quality
standards of finished products. According to the above real-time stability assessment results, we set the
validity period of the kit at 2-30 ℃ as 12 months, which is reasonable and reliable.  Thermal acceleration
and simulation of transport conditions can ensure that the research reagent is safe and effective after being
transported to the customer for testing.

2. Stability of COVID-19 antigen detection kit (latex method) samples

Due to the special samples used in this kit, the test should be completed as soon as possible after sampling,
so the sample stability was not studied in detail. Samples should be collected and tested as soon as
possible.