Gentaur 2 Antigen Test Kit (Colloidal Gold)

Contents

The clinical test of SARS-CoV-2 Antigen Test Kit (Colloidal Gold) was carried out; a
comparative study was conducted, and clinical diagnostic criteria of pneumonia
caused by novel coronavirus and the result of disease progression were used as the
comparison method. This study is a completely retrospective study. Clinical trials
were conducted in Shenzhen Center for Disease Control and Prevention (Shenzhen
CDC) and Shenzhen Engineering Laboratory for High-throughput Gene Sequencing
of Pathogens, Shenzhen Children’s Hospital. Testing was performed in August 2020.

L. Gevaert,Bio-engineer RUG 1996

1 General

1.1 The source, biology and physicochemical properties of the measured object
A novel coronavirus is a β genus coronavirus. It is enveloped and has round or elliptic
particles, usually pleomorphic, with a diameter of 60-140nm. Its genetic
characteristics were significantly different from sars-cov and mers-cov. At present, its
homology with of bat sars-like coronavirus (bat-sl-covzc45) is more than 85%.
At present, the infection source is mainly from novel coronavirus infected patients.
Asymptomatic infections may also be a source of infection.
Respiratory droplets and close contact is the main route of transmission. It is possible
to spread by aerosol in a relatively closed environment when exposed to a high
concentration of aerosol for a long time. Since novel coronavirus can be isolated from
feces and urine, note that feces and urine cause aerosol or contact transmission to the
environment.
The crowd is generally susceptible. Based on the current epidemiological
investigation, the incubation period is 1-14 days, most of which are 3-7 days. With
fever, dry cough, fatigue as the main performance. A few patients were accompanied
by nasal congestion, runny nose, sore throat, myalgia, and diarrhea. Severe patients
developed dyspnea and/or hypoxemia one week after the onset of the disease, and in
severe cases, they rapidly progressed to acute respiratory distress syndrome, septic
shock, refractory metabolic acidosis, coagulation dysfunction and multi-organ failure.
It is important to note that the course of severe and critical patients can be moderate or
low fever, or even no obvious fever.

1.2 Clinical intended use

Genrui SARS-CoV-2 Antigen Test Kit (Colloidal Gold) is an
immunochromatographic assay for rapid, qualitative detection of severe acute
respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen from the oropharyngeal
swab or nasopharyngeal swab. The test is to be used as an aid in the diagnosis of
coronavirus infection disease (COVID-19), which is caused by SARS-CoV-2.

The test provides preliminary test results. Negative results cannot exclude
SARS-CoV-2 infection and they cannot be used as the sole basis for treatment or
other management decision.
For in vitro diagnostic use only. For professional use only.

1.3 Test Principle

This product uses highly specific antibody-antigen reaction and colloidal gold
immunochromatographic technology. The reagent contains anti- SARS-CoV-2
monoclonal antibodies pre-fixed on the test area (T) on the membrane and anti
SARS-CoV-2 monoclonal antibody gold-labeled conjugate labeled on the gold label
pad.
During the test, the processed sample to be tested is dropped into the reagent loading
place. When the sample contains SARS-CoV-2 antigen, the SARS-CoV-2 antigen in
the sample is first combined with the anti- SARS-CoV-2 antibody labeled with
colloidal gold, and then the conjugate is chromatographed on the membrane, and it is
combined with another anti- SARS-CoV-2 antibody pre-immobilized on the
membrane. When the anti- SARS-CoV-2 monoclonal antibody binds, a purple-red
band will appear in the test area (T). If there is no SARS-CoV-2 antigen in the sample,
there will be no purple-red band in the test area (T). Regardless of whether the
SARS-CoV-2 antigen is present in the sample, a purple-red band will appear in the
quality control area (C). The purple-red band in the quality control area (C) is the
standard for judging whether there are enough samples and whether the
chromatography process is normal, and it also serves as an internal control standard
for reagents.

2 Research purposes

Qualitative detection of severe acute respiratory syndrome coronavirus 2
(SARS-CoV-2) antigen was performed on oropharyngeal swab or nasopharyngeal
swab specimen from confirmed/excluded SARS-CoV-2 cases with assessment reagent.
The test results were compared with the clinical diagnostic criteria of covid-19 and
the determination results of disease progression, and verify the clinical performance
and effectiveness of the product.

3 Experiment management

3.1 Laboratory quality control
Formulated and strictly implemented the standard operating procedures for clinical
testing projects, established laboratory quality control procedures, and regularly
trained laboratory staff on quality control, statistics and various clinical testing
techniques.
3.2 Statistics/data management
(1) The original data generated from the clinical trial shall be recorded and reviewed
by the operator in a complete and accurate manner to ensure the reliability of the data
and ensure that the conclusions in the clinical trial are derived from the original data.
(2) The process of statistical analysis of clinical trial data and the expression of its
results adopt standard statistical methods, and the special person who is familiar with
medical statistics is responsible for it.
4 Experiment design
4.1 Description of the experiment
The clinical diagnosis criteria of novel coronavirus pneumonia and the judgment of
disease progression were selected as comparative methods for comparative study to
verify the clinical performance of this product. This study is a clinically validated
study using blind methods.
4.2 Experiment design and test method selection
(1) Sample size
According to the diagnostic criteria for novel coronavirus pneumonia; the sample
oropharyngeal swab or nasopharyngeal swab confirmed cases no less than 60 and
excluded cases no less than 200.
(2) Sample selection basis, selection criteria, exclusion criteria and exclusion
criteria
1) Selection criteria
The enrolled population of the clinical trial should be the applicable population
of the product, which should be suspected cases of novel coronavirus pneumonia. For
the definition of “suspected cases”, refer to the “Novel Coronavirus Pneumonia
Diagnosis and Treatment Program (Trial Version 7)”.
The enrolled population of clinical trials should be able to represent various
types of populations that suitable for the product, including confirmed cases of
pneumonia (including partial recovery cases) and excluded cases of novel coronavirus
infection.
a. According to the clinical diagnosis, refer to the sample storage period of the
product manual, and select oropharyngeal swab or nasopharyngeal swab samples that
meet the requirements.
b. There is no limitation on gender and age. Select oropharyngeal swab or
nasopharyngeal swab samples of confirmed and excluded cases of novel coronavirus
pneumonia.
2) Exclusion criteria
a. Those that did not meet the selection criteria and were selected by mistake;
b. Those that do not meet the sample storage requirements;
c. The sample information records are confusing or ambiguous.
3) Elimination criteria
a. Those that fail to meet the selection criteria and were selected by mistake and
those who meet the exclusion criteria but not excluded;
b. Incorrect results due to operational errors and quality control failures.
(3) Sample collection, storage method, and transfer method

1) Sample collection
All samples come from the remaining samples of clinical testing. According to
sample selection basis and selection criteria, swab samples are collected in clean, dry,
waterproof containers that free of transmission media, preservatives, and detergents.
Perform blind coding according to the blind coding and solve blindness procedures,
and complete the sample selection.
2) Sample storage

The sample should be used as soon as possible after collection. If it cannot be
used immediately, it must be stored at 2-8°C within 3 days. For long-term storage, it
must be stored frozen below -70°C.
(4) Establishment of comparison method
The clinical diagnosis of novel coronavirus-infected pneumonia and the judgment
results of disease progression (Reference PCR results) were selected as comparative
method for comparative study.
(5) All products used in clinical trials
Reagent Test reagents
Name SARS-CoV-2 Antigen Test Kit (Colloidal Gold)
Specification / Model 25 T/Kit
Manufacturer Genrui Biotech Inc.
LOT 20200307
Expiration date 20210906
Storage Condition 2-30℃
(6) Quality control method
1) Operation training
Before the start of this clinical trial, the manufacturer conducts training and
pre-experiment on the trial researchers to enable the trial researchers to achieve the
following goals after training:
a. Familiar and understand the clinical trial plan, familiar with the “Instruction”
of the reagents used, and master the operation steps.
b. Be familiar with the safety instructions, precautions and relevant warnings in
the “Instruction ” of the reagents used.
c. Confirm all the functions of the reagents used. If there are any abnormal
conditions that do not conform to the “Instruction”, the sponsor should be notified in
time, and the test can be carried out after the abnormalities are eliminated.
d. Implement unified recording methods and judgment standards.

2) Process control

a. The researcher should follow the requirements of the clinical trial record form
and record the contents of the trial truthfully, in detail, and carefully to ensure that the
content of the clinical trial record form is complete, true and reliable.
b. All observations and findings during this trial should be verified to ensure the
reliability of the data and ensure that the conclusions in the clinical trial are derived
from the original data.
c. In order to eliminate the influence of subjective deviations and personal
preferences that may appear in the consciousness of the operator on the test results,
this test uses a blind design.
3) Test control
The test is carried out in strict accordance with the instruction.
4) Security control
a. All human materials in this product have been determined to be free of
HBsAg, HIV antibody, Treponema pallidum antibody and HCV antibody.
b. The clinical samples tested by this product are oropharyngeal swab or
nasopharyngeal swab. The test researchers should strictly follow the requirements of
the “National Clinical Inspection Operation Rules” and the instructions for use, wear
masks and latex gloves to prevent biological contamination.
c. The product is not in direct contact with the patient during use. The samples
tested are the remaining waste samples routinely tested by the test institute, and the
data measured by this product are only used for clinical trials, not as a clinical test
report issued by the hospital. There is no risk.
(7) Clinical performance evaluation method
Using clinical diagnostic criteria as controls, the clinical sensitivity, clinical
specificity, and confidence interval of the detection reagent test results were
calculated.
(8) Evaluation criteria of clinical performance statistical method
1) Statistical method
The results are summarized in the form of a 2 × 2 table, and the clinical
sensitivity, clinical specificity, and confidence interval are calculated accordingly. A
two-sided test was used, and the statistically significant test level α was set at 0.05,
and the parameter confidence interval was estimated to use 95% confidence interval.
2) Evaluation criteria
a. Clinical sensitivity
The results are summarized in the form of a 2 × 2 table, and the clinical
sensitivity of the detection results is ≥85%.
b. Clinical specificity
The results are summarized in the form of a 2 × 2 table, and the clinical
specificity of the detection results is ≥95%.
5 Analysis of clinical trial results
Using clinical diagnostic criteria as controls, the clinical sensitivity, clinical
specificity, and confidence interval of the detection results were calculated.
PCR method
Subtotal
Pos Neg
SARS-CoV-2 Antigen
Test Kit (Colloidal Gold)
Pos 59 2 61
Neg 8 203 211
Subtotal 67 205 272
Positive Percent Agreement: 88.06% (95%CI: 77.82% – 94.70%)
Negative Percent Agreement: 99.02% (95%CI: 96.52% – 99.88%)
Overall Percent Agreement: 96.32% (95%CI: 93.34% – 98.22%)
6 Discussion and conclusion
To conduct clinical trials on the SARS-CoV-2 Antigen Test Kit (Colloidal Gold)
produced by Genrui Biotech Inc., the clinical diagnosis of novel coronavirus
pneumonia and the judgment results of disease progression were used as comparative
method for comparative study.

 

Statistical analysis results:

The clinical sensitivity of the detection results is 59/67: 88.06% (77.82% – 94.70%);
the clinical specificity of the detection results is: 203/205: 99.02% (96.52% – 99.88%).
Overall Percent Agreement is 262/272:96.32% (93.34% – 98.22%).

Statistical analysis results show that the SARS-CoV-2 Antigen Test Kit (Colloidal
Gold) produced by Genprice Inc. meets the clinical needs.

Lieven Gevaert