Wondfo SARS-CoV-2 Antibody Test (Lateral Flow Method)是一种免疫层析法，用于快速、定性地检测人全血、血清或血浆样本中严重急性呼吸综合征冠状病毒2型(SARS-CoV-2) IgG/IgM抗体。本试验将有助于诊断由SARS-CoV-2引起的冠状病毒感染疾病(COVID-19)。
试验提供了初步的试验结果。阴性结果不能排除SARS-CoV-2感染，它们不能作为治疗或其他管理决策的唯一依据。
仅用于体外诊断。只供专业人士使用。
Wondfo SARS-CoV-2 Antibody Test (Lateral Flow Method)是基于捕获免疫分析法的原理，检测人全血、血清和血浆中的SARS-CoV-2 IgG/IgM抗体。当试样加入到测试装置中时，试样通过毛细作用被吸收到装置中，与SARS-CoV-2抗原-染料偶联物混合并流经预涂膜。
当标本SARS-CoV-2抗体水平以上目标截止(测试的检测极限),捕获抗体绑定到antigen-dye共轭反人类免疫球蛋白抗体和反μ链抗体固定在测试区域(T)的设备,这产生一个彩色测试带,表明一个积极的结果。当样本中的SARS-CoV-2抗体水平为零或低于目标截止值时，该装置的测试区域(T)没有可见的彩色带。这表示结果是否定的。
提供的材料
- 20个独立密封袋，每个袋内装:□1盒测试盒
1个干燥袋
- 2 .一次性滴管20支检测缓冲液(1* 6ml)使用说明的
需要但未提供的材料
- 样本收集容器
- 离心机(用于血清/血浆样本)
3.计时器
- 个人防护装备，如防护手套、医用
面罩，护目镜和实验服。
- 适当的生物危害废物容器和消毒剂。
- 本试剂用于检测人全血、血浆、血清样本中SARS-CoV-2抗体。
- 这是一种定性分析试剂。它不是用来测定SARS-CoV-2抗体的定量浓度的。
3.测试的准确性取决于样品采集过程。样品采集不当、储存不当或反复冻融会影响检测结果。
- 本试剂的检测结果仅供临床参考，只有在所有临床及实验室检查结果均经过评估后才能确诊。
- 受抗体检测试剂方法的限制，阴性检测结果建议使用核酸检测或病毒培养鉴定方法进行复核确认。
- 阳性检测结果并不排除与其他病原体合并感染的可能。该试剂的阴性结果可由以下原因引起:1)样品采集不当，样品转移不当或
处理时，样品中的病毒效价过低;
2) SARS-CoV-2抗体水平低于检测限
的测试。
3)病毒基因的变异可能导致抗体的改变
决定因素。
获取596例临床病例样本进行检测，其中包括361例确诊病例样本*和235例确诊排除病例样本*，然后将Wondfo SARS-CoV-2抗体检测(侧流法)与确诊病例样本的检测结果进行对比。结果两者之间的敏感性和特异性
下面展示了两种方法。
*确诊病例为根据治疗方案诊断的患者。
*经证实的排除性护理经PCR阴性结果鉴定。
结果分析:
敏感性:86.43% (95%CI: 82.41%~89.58%)特异性:99.57% (95%CI: 97.63%~99.92%)
总一致性:91.61% (95%CI: 89.10%~93.58%)
b .大
采用Wondfo SARS-CoV-2抗体试验(侧流法)对患者标本进行检测。结果显示无交叉反应。
阴性值和阳性值100%正确识别
时间。
- 之间的运行精度是用三种不同的方法来确定的
在3个不同批次的试验装置中含有不同浓度抗体的标本。阴性和阳性结果再次被100%正确识别。
参考书目
朱娜，博士，张定宇，博士，王文玲，等(2020)。一种来自中国肺炎患者的新型冠状病毒，2019年。新英格兰医学杂志。
[2]陈旺，Peter W Horby, Frederick G Hayden, George F Gao。(2020)。全球健康关注的新型冠状病毒爆发。《柳叶刀》，395(10223)，470-473。
黄朝林，叶明，等(2020)。武汉地区2019年新型冠状病毒感染者的临床特征。《柳叶刀》，395(10223)，497-506。
陈南山，周敏，董轩，等(2020)。武汉地区2019年新发冠状病毒肺炎99例流行病学及临床特征的描述性研究。《柳叶刀》，395(10223)，507-513。
2020年1月12日，世界卫生组织:当怀疑新冠状病毒(nCoV)感染时对严重急性呼吸道感染的临床管理:临时指南。
指数的符号
试剂
临床病例
总计
确诊病例样本
排除案例样本
Wondfo SARS-CoV-2抗体检测(侧流法)
积极的
312
1
313
负
49
234
283
总计
361
235
596
副流感病毒病毒抗体
甲型流感抗体
乙型流感抗体
肺炎衣原体抗体
肺炎支原体抗体
腺病毒抗体
呼吸道合胞体病毒抗体
乙型肝炎表面抗体
丙型肝炎病毒抗体
梅毒螺旋体抗体
艾滋病病毒抗体
EB病毒抗体
麻疹病毒抗体
巨细胞病毒抗体
肠病毒71型抗体
流行性腮腺炎抗体
水痘-带状疱疹病毒阳性样本
在体外
诊断使用
见使用说明
截止日期
每箱测试
生产日期
保持干燥
批号
制造商
避免阳光照射
2
存储2 ~ 30摄氏度之间
不重用
目录#
c干扰
Wondfo SARS-CoV-2抗体试验(侧流法)的检测结果不干扰以下浓度的物质:
物质
浓度
胆红素
250年μmol / L
血红蛋白
9 g / L
甘油三酸酯
15更易/ L
类风湿因子
80国际单位/毫升
抗核抗体效价
1:240
Anti-mitochondrial抗体(AMA)
80 U /毫升
小鼠免疫球蛋白
1000μg /毫升
d .精度
在运行精度是通过使用10个重复的三个不同的样品含有不同浓度的抗体。
INTENDED USE
Wondfo SARS-CoV-2 Antibody Test (Lateral Flow Method) is an
immunochromatographic assay for rapid, qualitative detection of
severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)
IgG/IgM antibody in human whole blood, serum or plasma sample.
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 don’t
preclude 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.
SUMMARY
On December 31, 2019, several cases of pneumonia in Wuhan City,
Hubei Province of China were reported to the World Health
Organization (WHO). The novel virus, now known as SARS-CoV-2
(previously known as 2019-nCoV), a RNA virus of the beta
coronavirus family, has since spread across China and to other
countries and territories. The WHO has named the disease caused by
SARS-CoV-2 as coronavirus disease 2019 (abbreviated
“COVID-19”).
PRINCIPLE
Wondfo SARS-CoV-2 Antibody Test (Lateral Flow Method) is based
on the principle of capture immunoassay for determination of
SARS-CoV-2 IgG/IgM antibodies in human whole blood, serum and
plasma. When the specimen is added into the test device, the
specimen is absorbed into the device by capillary action, mixes with
the SARS-CoV-2 antigen-dye conjugate and flows across the
pre-coated membrane.
When the SARS-CoV-2 antibodies level in the specimen is at or
above the target cutoff (the detection limit of the test), the antibodies
bound to the antigen-dye conjugate are captured by anti-human IgG
antibody and anti-human μ chain antibody immobilized in the Test
Region (T) of the device, and this produces a colored test band that
indicates a positive result. When the SARS-CoV-2 antibody level in
the specimen is zero or below the target cutoff, there is not a visible
colored band in the Test Region (T) of the device. This indicates a
negative result.
To serve as a procedure control, a colored line will appear at the
Control Region (C), if the test has been performed properly.
PRECAUTION
- This kit is for in vitro diagnostic use only.
- All specimens should be treated as capable of transmitting
diseases. Use appropriate precautions in the collection, handling,
storage and disposal of patient samples and used kit contents. And
follow biosafety level 2 or higher guidelines.
- Wear appropriate personal protective equipment (e.g. gowns,
gloves, eye protection) when handing the contents of this kit.
- Proper specimen collection storage and transport are critical to the
performance of this test.
- Discard after first use. The test cannot be used more than once.
- Do not touch the reaction area of test strip.
- Do not use test kit beyond the expiration date.
- Do not use the kit if the pouch is punctured or not well sealed.
- Testing should be applied by professionally trained staff working in
certified laboratories or clinics at which the sample(s) is taken by
qualified medical personnel.
- The test result should be interpreted by the physician along with
clinical findings and other laboratory test results.
- DISPOSAL OF THE DIAGNOSTIC: All specimens and the
used-kit has the infectious risk. The process of disposing the
diagnostic must follow the local infectious disposal law or
laboratory regulation.
MATERIAL
Material Provided
- 20 Individual sealed pouches, each pouch contains:
■ 1 x Test cassette
■ 1 x Desiccant pouch
- 20 disposable droppers
- Detection buffer (1*6 mL)
- Instructions for use
Material Required but Not Provided
- Specimen Collection Containers
- Centrifuge (for serum/plasma sample)
- Timer
- Personal protective equipment, such a protective gloves, medical
mask, goggles and lab coat.
- Appropriate biohazard waste container and disinfectants.
STORAGE AND STABILITY
- Store at 2~30ºC in the sealed pouch up to the expiration date
printed on the package. Do not freeze.
- The test cassette should be used within 1 hour after taking out from
the foil envelope. Buffer solution should be re-capped in time after
use.
- Keep away from sunlight, moisture and heat.
- Kit contents are stable until the expiration date printed on the outer
box.
SPECIMEN COLLECTION AND PREPARATION
The test can be performed with whole blood, serum and plasma.
For whole blood:
- Using standard phlebotomy procedure, collect a venipuncture
whole blood specimen using a blood collection tube with suitable
anticoagulant (containing EDTA, Heparin or Citrated sodium).
Other anticoagulants have not been validated and may give
incorrect result.
- It is recommended that whole blood specimen is tested at the time
- of specimen collection. If the specimens are not tested
immediately, they may be stored at 2ºC～8ºC for up to 7 days.
Prior to testing, mix the blood by gentle inversion several times, do
not freeze or heat whole blood specimens.
For Serum and Plasma:
- Using standard phlebotomy procedure, collect a venipuncture
whole blood specimen using a blood collection tube. If collecting
plasma use a blood collection tube containing suitable
anticoagulant (containing EDTA, Heparin or Citrated sodium).
Other anticoagulants have not been validated and may give
incorrect result.
- Centrifuge whole blood and separate the plasma from red blood
cell as soon as possible to avoid hemolysis.
- Test should be performed within 8 houres after the specimens have
been collected. Do not leave the specimens at room temperature
for prolonged periods. Serum or plasma specimens may be stored
at 2ºC～8ºC for up to 3 days prior to testing. Serum or plasma
specimens may be stored at -20ºC for up to 9 days.
Note: Bring specimens to room temperature before testing. Frozen specimens
must be completely thawed and mixed well prior to testing. Specimens should
not be frozen and thawed repeatedly. Severe hemolytic or heat-inactivated
specimens are not recommended.
- TEST PROCEDURE
Please read the instruction for use carefully before performing the
test.
- Allow the device, buffer and specimen to equilibrate to room
temperature (10ºC ～30ºC) prior to testing.
- Remove a test cassette from the foil pouch by tearing at the notch
and place it on a level surface.
- Transfer 10 μL of whole blood or serum or plasma specimen to the
sample well (small well) and then add 2-3 drops (80 μL) of buffer
solution to the buffer well (large well).
- As the test begins to work, you will see purple color move across
the result window in the center of the test device.
- Wait for 15 minutes and read the results. Do not read results after
20 minutes.
RESULT INTERPRETATION
Positive Result
Colored bands appear at both test line (T) and control line (C). It
indicates a positive result for the SARS-CoV-2 antibodies in the
specimen.
Negative Result
Colored band appear at control line (C) only. It indicates that the
concentration of the SARS-CoV-2 antibodies is zero or below the
detection limit of the test.
Invalid Result
No visible colored band appear at control line after performing the
test. The directions may not have been followed correctly or the test
may have deteriorated. It is recommended that the specimen be
re-tested.
QUALITY CONTROL
A procedural control is included in the test. A colored line appearing in
the control region (C) is considered an internal procedural control. It
confirms sufficient specimen volume, adequate membrane wicking
and correct procedural technique.
Good laboratory practice recommends the use of the control
materials. Users should follow the appropriate federal state, and local
guidelines concerning the frequency of assaying external quality
control materials.
LIMITATIONS OF PROCEDURE
- This reagent is designed to detect antibodies against SARS-CoV-2
in human whole blood, plasma, serum sample.
- This reagent is a qualitative assay. It is not designed to determine
the quantitative concentration of SARS-CoV-2 antibodies.
- The accuracy of the test depends on the sample collection process.
Improper sample collection, improper sample storage, or repeated
freezing and thawing of the sample will affect the test result.
- The test result of this reagent are for clinical reference only, a
confirmed diagnosis should only be made after all clinical and
laboratory findings have been evaluated.
- Limited by the method of antibody detection reagents, for negative
test results, it is recommended to use nucleic acid detection or
virus culture identification methods for review and confirmation.
- Positive test results do not rule out co-infections with other
pathogens. A negative result of this reagent can be caused by:
1) Improper sample collection, improper sample transfer or
handing, the virus titer in the sample is too low;
2) The level of SARS-CoV-2 antibodies is below the detection limit
of the test.
3) Variations in viral genes may cause changes in antibody
determinants.
PERFORMANCE CHARACTERISTICS
- Sensitivity and Specificity
596 clinical case samples which include 361 confirmed case
samples* and 235 confirmed excluded case samples*, were obtained
for testing, and then compared the test results between Wondfo
SARS-CoV-2 Antibody Test (Lateral Flow Method) and the confirmed
case samples. The results of sensitivity and specificity between the
two method are show below.
Confirmed cases were the patients diagnosed according to the
treatment plan.
* Confirmed excluded cares were identified by negative PCR results.
Results analysis：
Sensitivity: 86.43% (95%CI: 82.41%~89.58%)
Specificity: 99.57% (95%CI: 97.63%~99.92%)
Total consistent: 91.61% (95%CI: 89.10%~93.58%)
- Cross-reactivity
Specimens which tested positive with following various agents from
patients were investigated with Wondfo SARS-CoV-2 Antibody Test
(Lateral Flow Method). The results showed no cross reactivity.
- Interferences
The test result of Wondfo SARS-CoV-2 Antibody Test (Lateral Flow
Method) do not be interfered with the substance at the following
concentration:
Substance
Bilirubin
Hemoglobin
Triglyceride
Rheumatoid factors
Antinuclear antibody (ANA) titer
Anti-mitochondrial antibody (AMA)
Mouse IgG
Concentration
250 μmol/L
9 g/L
15 mmol/L
80 IU/mL
1:240
80 U/mL
1000 μg/mL
- Precision
- Within run precision was determined by using 10 replicates of three
different specimens containing different concentrations of antibody.
The negative and positive values were correctly identified 100% of
the time.
- Between run precision was determined by using the three different
specimens containing different concentrations of antibody in 3
different lots of test devices. Again negative and positive results
were correctly identified 100% of the time.
BIBLIOGRAPHY
[1] Na Zhu, Ph.D., Dingyu Zhang, M.D., Wenling Wang, Ph.D., et al.
(2020). A Novel Coronavirus from Patients with Pneumonia in
China, 2019. The New England Journal of Medicine.
[2] Chen Wang, Peter W Horby, Frederick G Hayden, George F Gao.
(2020). A novel coronavirus outbreak of global health concern. The
Lancet, 395(10223), 470-473.
[3] Chaolin Huang, Yeming, et al. (2020). Clinical features of patients
infected with 2019 novel coronavirus in Wuhan, China. The
Lancet, 395(10223), 497-506.
[4] Nanshan Chen, Min Zhou, Xuan Dong, et al. (2020).
Epidemiological and clinical characteristics of 99 cases of 2019
novel coronavirus pneumonia in Wuhan, China: a descriptive
study. The Lancet, 395(10223), 507-513.
[5] World Health Organization: Clinical management of severe acute
respiratory infection when Novel coronavirus (nCoV) infection is
suspected: Interim Guidance. 12 January, 2020.