CN111366525B - Method for rapidly detecting SARS-CoV-2 virus infection in isolated blood sample and application thereof - Google Patents
Method for rapidly detecting SARS-CoV-2 virus infection in isolated blood sample and application thereof Download PDFInfo
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- CN111366525B CN111366525B CN202010171734.1A CN202010171734A CN111366525B CN 111366525 B CN111366525 B CN 111366525B CN 202010171734 A CN202010171734 A CN 202010171734A CN 111366525 B CN111366525 B CN 111366525B
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a method for rapidly detecting SARS-CoV-2 virus infection in an isolated blood sample and application thereof, belonging to the technical field of biological medicine, the invention analyzes the position and corresponding quantity change of peripheral blood mononuclear cells based on FSC/SSC parameter change of flow cytometry to identify whether SARS-CoV-2 virus infection exists, and can be used for designing a specific detection instrument by combining with a photoelectric technology, and can find whether blood cells are infected by SARS-CoV-2 virus within 10 minutes. The analysis of the specific phenotypic molecular change of the specific mononuclear-macrophage of SARS-CoV-2 virus infected patients based on flow cytometry can be used for monitoring the infection condition.
Description
Technical Field
The invention belongs to the technical field of biological medicine, and relates to a method for rapidly detecting SARS-CoV-2 virus infection in an isolated blood sample and application thereof.
Background
2019 coronavirus disease (coronavirus disease 2019, CODV-19), and the virus causing the disease is named severe acute respiratory syndrome coronavirus 2(severe acute respiratory syndrome coronavirus 2, SARS-CoV-2 for short) [1 ]. SARS-CoV-2 is a novel coronavirus belonging to the genus beta of the same genus as SARS-CoV and MERS-CoV found previously, and has a cell membrane, and virus particles are circular or elliptical, mostly polymorphic, and have a diameter of 60-140 nM. SARS-CoV-2 is clearly distinguished from previously discovered SARS-CoV and MERS-CoV in terms of gene sequence, and homology with bat SARS-like coronavirus (bat-SL-CoVZC45) is currently found to be as high as 85% [2 ]. The population is generally susceptible to SARS-CoV-2, and based on epidemiological investigation, the latent period of the disease is about 1-14 days, and is mostly 3-7 days. The current laboratory detection of SARS-CoV-2 infection is mainly based on: (1) and (3) checking the etiology: viral nucleic acid is found in nasopharyngeal swab, sputum, secretion of lower respiratory tract, blood, excrement and other specimens by real-time fluorescent quantitative RT-PCR or genome sequencing technology. (2) Serological examination: detecting the specific IgM antibody and IgG antibody of the novel coronavirus. (3) Breast imaging: based on the early speckle images and the interstitial changes, and the characteristics of the ground glass images are screened. However, the above method still has obvious disadvantages: the detection rate and accuracy of RT-PCR are affected by factors such as the quality of a test sample, the virus titer and the like; serological detection has certain window period requirements; chest imaging has a low detection rate for atypical patients.
In 2003, researchers identified angiotensin-converting enzyme 2 (ACE 2) ACE2 as a functional receptor infected by SARS-CoV, SARS-CoV-2 and human coronavirus NL63 virus based on co-immunoprecipitation technology[3-6]. In vitro experiments demonstrated that the novel coronavirus was able to infect cells as long as the cells expressed ACE 2. The Spike protein of the virus interacts with ACE2 on the cell surface, and further internalizes through ACE2 to cause endocytosis of virus particles, induces fusion of the virus and host cells, and causes SARS-CoV infection. ACE2 is an exopeptidase which catalyzes the conversion of angiotensin I to angiotensin- (1-9) or angiotensin II to angiotensin- (1-7) and down-regulates the activated renin-angiotensin system[7,8]. ACE2 as SARS-CoV virus receptor function and degradation of Ang IICatalytic activity is not directly related mechanistically, but ACE 2-mediated degradation of Ang II is also important for lung protection from the pathogenesis of pneumonia caused by SARS-CoV infection, since ACE2 loss may exacerbate the symptoms of acute lung injury[9]。
A Flow Cytometer (FCM) is an advanced biomedical measuring instrument developed on the basis of cross fusion of laser technology, fluorescence labeling technology, antibodies, photoelectric technology, and the like. Flow cytometry is a technique for rapid, quantitative, and multi-parameter analysis of cells or other biological particles using flow cytometry. The cell suspension of the sample to be detected is wrapped and restrained by sheath liquid under the fine liquid flow and pressure control of a flow cytometer by one or more special fluorescent labels, and the cells are arranged in a single row and are ejected out from a nozzle of a flow chamber at high speed to form a cell liquid column. When the liquid column passes through the optical detection area, the cells in the liquid column generate forward scattered light (FSC) and side scattered light (SSC) and fluorescence under the irradiation of an incident laser beam, which respectively reflect the cell size (FSC), granularity (SSC) and intracellular and extracellular characteristics, and the cells can be classified according to the characteristics[10,11]. This significant advantage of flow cytometry has led to its unique advantages in the classification and analysis of blood and immune cells, among others.
Disclosure of Invention
The invention aims to provide a method for rapidly detecting SARS-CoV-2 virus infection in an isolated blood sample and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:
the invention discloses a method for rapidly detecting SARS-CoV-2 virus infection in an isolated blood sample, which monitors the isolated blood sample by flow cytometry, inspects the change of the form and the position of a monocyte in the blood sample to be detected by FSC/SSC, and judges the SARS-CoV-2 virus infection result.
Preferably, in a blood sample infected with SARS-CoV-2 virus, the number of cells in the P1 gate is reduced and the number of cells in the P2 gate is increased;
wherein the P1 gate represents a normal monocyte subpopulation; the P2 phylum is a characteristic cell subset of SARS-CoV-2 virus.
Further preferably, the cell subpopulation of the P2 gate is a monocyte derived macrophage.
Still further preferably, the monocyte-derived macrophage is capable of expressing cell-specific marker molecules CD11b, CD68, CD80 and CD 163.
The invention also discloses the application of the angiotensin converting enzyme 2 as a blood mononuclear cell SARS-CoV-2 virus detection target.
Preferably, the blood sample is monitored by flow cytometry in vitro, and the position change of the expression of angiotensin converting enzyme 2 of the mononuclear cells in the blood sample to be tested is examined by FSC/SSC to judge the SARS-CoV-2 virus infection result.
Preferably, in the blood sample infected by SARS-CoV-2 virus, the angiotensin converting enzyme 2 shows high expression in monocytes both in the P1 gate and in the P2 gate;
wherein the P1 gate represents a normal monocyte subpopulation; the P2 phylum is a characteristic cell subset of SARS-CoV-2 virus.
Preferably, the human monocyte cell line U937, the human monocyte cell line THP1 and the mouse macrophage cell line RAW264.7 can express SARS-CoV-2 virus and the receptor angiotensin converting enzyme 2 of SARS-CoV-2 virus.
Compared with the prior art, the invention has the following beneficial effects:
the method disclosed by the invention is used for analyzing the position and the corresponding quantity change of peripheral blood mononuclear cells based on the FSC/SSC parameter change of flow cytometry to identify whether the peripheral blood mononuclear cells are infected by SARS-CoV-2 virus, can be used for designing a specific detection instrument by combining a photoelectric technology, and can observe whether the blood cells are infected by SARS-CoV-2 virus within 10 minutes. The analysis of the specific phenotypic molecular change of the specific monocyte-macrophage of SARS-CoV-2 virus infected patients based on flow cytometry can be used for monitoring the infection condition.
Drawings
FIG. 1 is a graph of peripheral blood results of flow cytometry FSC/SSC parameter analysis of healthy humans and SARS-CoV-2 virus infected patients; wherein A is a representative healthy human peripheral blood flow cytometry FSC/SSC graph; b is a representative SARS-CoV-2 virus infection patient peripheral blood flow cytometry FSC/SSC map; c is the statistical analysis result of the ratio of cells in 10 healthy people and 28 SARS-CoV-2 virus infected patients in peripheral blood flow cytometry FSC/SSC graph P1 and P2;
FIG. 2-1 is a flow cytometric analysis of the distinct cellular immunophenotype of the P2 subgroup significantly increased in peripheral blood of SARS-CoV-2 virus infected patients; wherein A is P1 and P2 gates under the peripheral blood FSC/SSC parameter of a patient infected by SARS-CoV-2 virus; b is CD11B +; c is CD14 +; d is CD68 +; e is CD80 +; f is CD163 +;
FIG. 2-2 is a comparison of statistics for B-F in FIG. 2-1;
FIG. 3 is a flow cytometry analysis of ACE2 expression in U937, THP1 and RAW264.7 cells; wherein, A is high expression ACE2 of U937 cell line; b is expression of high expression ACE2 of THP1 cell line; c is mouse RAW264.7 cell line high expression ACE 2;
FIG. 4 is a diagram of flow cytometry analysis of ACE2 expression of cells in a healthy human peripheral blood mononuclear cell subset (P1 gate) and a SARS-CoV-2 virus infected patient marker P2 gate;
FIG. 5 is a flow cytometry analysis of ACE2 expression from SARS-CoV-2 virus infected patient peripheral blood mononuclear macrophage.
Detailed Description
In order to make the technical solutions of the present invention better understood by those skilled in the art, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, shall fall within the scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
The invention is described in further detail below with reference to the accompanying drawings:
1. cell population with altered location in peripheral blood of SARS-CoV-2 virus infected patient found under FSC and SSC parameters of flow cytometry
1.1 Experimental methods:
(1) the confirmed hospitalized SARS-CoV-2 virus infected patient is from the eighth hospital of Ann city, Shaanxi province, China, and the test sample is from the residual blood of patient blood routine detection, K3 EDTA anticoagulation.
(2) 50 μ L of peripheral blood of healthy volunteers and SARS-CoV-2 virus-infected patients were taken into flow tubes, 450 μ L of hemolysin (BD Co., Ltd., product No. 349202) was added to each tube, and the mixture was vortexed and left to stand for 5 minutes.
(3) 3-4mL of PBS (containing 0.5% BSA and 0.1 sodium azide) was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
(4) The cell pellet was vortexed and the cells resuspended with 500. mu.L PBS (containing 0.5% BSA and 0.1 sodium azide).
(5) Flow cytometry was used for detection, focusing on the monocyte subpopulations under FSC and SSC parameters.
For the BD cantonii flow cytometer, the FSC voltage used is around 350 and the SSC voltage around 385.
(6) The results were analyzed using FlowJo 7.6.1 software.
1.2 Experimental results:
results referring to FIG. 1, the P1 gate in FIG. 1 represents a normal monocyte subpopulation, and the P2 gate is a new characteristic cell subpopulation in SARS-CoV-2 virus-infected patients. As can be seen in FIG. 1, normal human monocytes (monocytes) are predominantly located within the P1 gate (Mean. + -. SEM of 4.31. + -. 0.42), with cell numbers in the P2 gate ranging between 0-0.4% (Mean. + -. SEM of 0.1653. + -. 0.04242); the number of cells in P2 gate of SARS-CoV-2 virus infected patient is increased significantly (Mean + -SEM is 2.241 + -0.1665), the number of cells in P1 gate of SARS-CoV-2 virus infected patient is decreased (Mean + -SEM is 1.370 + -0.2045), and the cells in P2 gate are proved to accord with the characteristics of macrophage (macrocage) by the results of subsequent experiments. There was significant statistical significance between the changes in cell numbers in the P1 and P2 populations of patients and healthy persons (P <0.0001, unpaired T test results).
2. Flow cytometry characterization of monocyte subpopulations in peripheral blood and positionally altered cell populations of SARS-CoV-2 virus infected patients
2.1 Experimental methods:
(1) confirmed hospitalized SARS-CoV-2 virus infected patients were obtained from the eighth Hospital, Western Ann, and the test samples were obtained from patient blood, routinely tested for residual blood, anticoagulated with K3 EDTA.
(2) Each patient took 3 flow tubes and added 50 μ L of peripheral blood of SARS-CoV-2 virus infected patients to the flow tubes, wherein:
1 tube without antibody as negative control;
1 tube is added with the combined antibodies of CD11b-PerCP-Cy5.5, CD14-PE, CD16-APC-Cy7, CD68-APC and CD80-FITC, and each antibody is added with 0.5 mu L;
1 tube is added with the combined antibodies of CD11b-PerCP-Cy5.5, CD14-PE, CD16-APC-Cy7, HLA-DR-FITC, CD68-APC and CD163-PE-Cy7, and each antibody is added with 0.5 mu L;
the antibodies are all from Biolegend and have the cargo numbers 101230, 301806, 302018, 333810, 305205, 307603 and 326514 respectively. Vortex and mix well, incubate 15 minutes at room temperature in the dark.
(3) 450 μ L of hemolysin (BD company, product number 349202) specific for flow cytometry was added to each tube, vortexed, and allowed to wait for 5 minutes.
(4) 3-4mL of PBS (containing 0.5% BSA and 0.1 sodium azide) was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
(5) The cell pellet was vortexed and the cells resuspended with 500. mu.L PBS (containing 0.5% BSA and 0.1 sodium azide).
(6) Flow cytometry was used for detection, focusing on the monocyte subpopulations under FSC and SSC parameters.
For the BD cantonii flow cytometer, the FSC voltage used is around 350 and the SSC voltage around 385.
(7) The results were analyzed using FlowJo 7.6.1 software.
2.2 Experimental results:
the macrophage differentiation characteristics of the monocytes which are shifted to the right in the peripheral blood of the SARS-CoV-2 virus-infected patients are shown in FIG. 2-1 and FIG. 2-2, and the result shows that the cells of the group express strong CD11b, CD68 and CD163, and express CD14 and CD80 simultaneously, which accord with the macrophage immunophenotype, and the increased P2 subgroup cells which are shown in the patients are mononuclear-derived macrophages.
3. Flow cytometry analysis shows that human THP-1, U937 monocyte cell line and mouse RAW264.7 macrophage cell line highly express ACE2
3.1 Experimental methods:
(1) respectively culturing human monocyte THP-1 and U937 cells in RPMI160 culture medium (supplemented with 10% fetal calf serum and double antibody); mouse RAW264.7 macrophage cell line was cultured in high-glucose DMEM medium (supplemented with 10% fetal bovine serum, without added diabody).
(2) The THP-1, U937 and RAW264.7 cells were collected in logarithmic growth phase into flow tubes, about 50 ten thousand cells per tube (2 tubes per cell, with 1 tube without primary antibody as a negative control).
(3) Centrifuge at 1500rpm for 5 minutes and discard the supernatant.
(4) The cell mass was vortexed, 3-4mL of PBS was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
3.2 results of the experiment
Results see fig. 3, in fig. 3, indirect staining: red represents fluorescent marker 2 anti-staining (negative control); blue represents staining results with primary anti-ACE 2 and fluorescent secondary antibody. MFI is the mean fluorescence intensity. From A, B and C in FIG. 3, it can be seen that the U937 cell line, the THP1 cell line and the mouse RAW264.7 cell line all highly expressed ACE 2.
4. Discovery of expression of ACE2 by peripheral blood mononuclear cell of healthy human through flow cytometry
4.1 Experimental methods:
(1) collecting peripheral blood of healthy volunteers to K3EDTA anticoagulation tube.
(2) 50 μ L of peripheral blood was taken from each person into flow tubes (2 tubes per person, 1 tube without primary antibody as negative control) and 0.2 μ L of rabbit anti-human ACE2 primary antibody (abcam, cat # ab87436) was added. Revolve and mixing. Staining was carried out at room temperature for 15 minutes in the dark.
(3) 450 μ L of hemolysin (BD corporation, product number 349202) specific for flow cytometry was added to each tube, vortexed, and allowed to wait for 5 minutes.
(4) 3-4mL of PBS (containing 0.5% BSA and 0.1 sodium azide) was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
(5) mu.L of PBS (containing 0.5% BSA and 0.1 sodium azide) was added to each tube, followed by 0.1. mu.L of goat anti-rabbit IgG-AF488 fluorescently labeled antibody, and 0.5. mu.L of CD14-PE (Biolegend, cat. 301806). Vortex and mix well, incubate 15 minutes at room temperature in the dark.
(6) 3-4mL of PBS (containing 0.5% BSA and 0.1 sodium azide) was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
(7) Repeating the previous step.
(8) The cell pellet was vortexed and the cells resuspended with 500. mu.L PBS (containing 0.5% BSA and 0.1 sodium azide).
(9) Detecting by using a flow cytometer.
For the BD cantonii flow cytometer, the FSC voltage used is around 350 and the SSC voltage around 385.
(10) The results were analyzed using FlowJo 7.6.1 software.
4.2 Experimental results:
referring to fig. 4, a total of 5 healthy persons were tested and the results showed that: healthy human peripheral blood mononuclear cells express strong ACE 2. ACE2 is expressed in most monocytes, whether gated by monocyte subpopulations or by the monocyte specific marker CD 14. Among them, monocytes within the P1 gate (normal position) expressed higher ACE2 than at the P2 position.
5. ACE2 is expressed by mononuclear cells and macrophages in peripheral blood of SARS-CoV-2 virus infected patient
5.1 Experimental methods:
(1) the confirmed hospitalized SARS-CoV-2 virus infected patient is from the eighth hospital in Xian city, the test sample is from the patient blood to conventionally detect residual blood, K3 EDTA anticoagulation;
(2) each patient took 2 tributary tubes and added 50. mu.L of SARS-CoV-2 virus infected patient peripheral blood into the flow tubes: in 1 of these samples, 0.2. mu.L of rabbit anti-human ACE2 primary antibody (abcam, cat. ab87436) was added to the sample in the other 1. Vortex and mix evenly. Dyeing for 15 minutes at room temperature in a dark place;
(3) 450 μ L of hemolysin (BD company, product number 349202) specific for flow cytometry was added to each tube, vortexed and mixed, and the mixture was allowed to wait for 5 minutes;
(4) 3-4mL of PBS (containing 0.5% BSA and 0.1 sodium azide) was added, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded.
(5) mu.L of PBS (containing 0.5% BSA and 0.1 sodium azide) was added to each tube, followed by 0.1. mu.L of goat anti-rabbit IgG-AF488 fluorescently labeled antibody, and 0.5. mu.L of CD14-PE (Biolegend, cat. 301806). Vortex, mix evenly, incubate 15 minutes in dark at room temperature;
(6) 3-4mL PBS (containing 0.5% BSA and 0.1 sodium azide) was added to each tube, centrifuged at 1500rpm for 5 minutes, and the supernatant was discarded;
(7) repeating the previous step;
(8) vortex the cell pellet, add 500 μ Ι _ PBS (containing 0.5% BSA and 0.1 sodium azide) to resuspend the cells;
(9) detection using flow cytometry
Using a BD cantonii flow cytometer as an example, the FSC voltage used is around 350, and the SSC voltage is around 385;
(10) the FlowJo 7.6.1 software analyzed the results.
5.2 Experimental results:
see FIG. 5 for a representative result of 5 patients infected with SARS-CoV-2 virus. The above results show that: monocytes from SARS-CoV-2 virus infected patients express strong ACE 2. Whether the gate was set by monocyte subpopulation or by monocyte-specific marker CD14 cycle, ACE2 was shown to be expressed in most monocytes and cells within P1 and P2 showed high expression.
In conclusion, the method disclosed by the invention proves that the monocyte and macrophage cell line of human and mouse and the peripheral blood mononuclear macrophage system of human express SARS-CoV and SARS-CoV-2 virus receptor ACE2 through experiments, which shows that the mononuclear macrophage can be attacked by SARS-CoV and SARS-CoV-2 virus infection. The position of some or all monocytes in the peripheral blood of a human infected with SARS-CoV-2 virus shifts to the right under the FSC/SSC parameters of the flow cytometer. The rightward shift of peripheral blood mononuclear cells indicates an increase in the volume of this fraction of mononuclear cells. Further analysis by flow cytometry phenotypic analysis proves that the part of the monocytes which are shifted to the right in position express macrophage marker molecules and are differentiated into macrophage-like cells. Based on the characteristic change of peripheral blood mononuclear cells, which is found by flow cytometry and is different from normal human, the method can be further combined with a photoelectric detection technology to judge whether the blood cells are infected by SARS-CoV-2 virus.
The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical solution according to the technical idea proposed by the present invention falls within the protection scope of the claims of the present invention.
The present invention relates to the following references:
【1】 World health organization official website:
https://www.who.int/zh/emergencies/diseases/novel-coronavirus-2019/technical- guidance/naming-the-coronavirus-disease-(covid-2019)-and-the-virus-that-causes-it.
【2】Cheng AC,Williamson DA.An outbreak of COVID-19caused by a new coronavirus:what we know so far.Med J Aust.2020Mar 8.doi:10.5694/mja2.50530.
【3】Weiss SR,Navas-Martin S.Angiotensin-converting enzyme 2--a new cardiac regulator.Microbiol Mol Biol Rev.2005,69(4):635-664.
【4】Kuba K.,et al.Trilogy of ACE2:A peptidase in the renin–angiotensin system, a SARS receptor,and a partner for amino acid transporters.Pharmacology& Therapeutics,2010,128(1):119-128.
【5】Xu,X.,et al.Evolution of the novel coronavirus from the ongoing Wuhan outbreak and modeling of its spike protein for risk of human transmission.Journal of SCIENCE CHINA Life Sciences,2020.
【6】Hoffmann M,et al.SARS-CoV-2Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor.Cell.2020,pii: S0092-8674(20):30229-30224.
【7】Donoghue M,et al.A novel angiotensin-converting enzyme-related carboxypeptidase(ACE2)converts angiotensin I to angiotensin 1-9.Circ.Res.2000,87 (5):E1-9.
【8】Keidar S,Kaplan M,Gamliel-Lazarovich A.ACE2 of the heart:From angiotensin I to angiotensin(1-7).Cardiovasc Res.2007,73(3):463-469.
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Claims (3)
1. a method for rapidly detecting SARS-CoV-2 virus infection in an isolated blood sample is characterized in that the isolated blood sample is monitored by flow cytometry, the change of the form and the position of a monocyte in the blood sample to be detected is inspected by FSC/SSC, and the SARS-CoV-2 virus infection result is judged;
in the blood sample infected by SARS-CoV-2 virus, the number of cells in P1 gate is reduced, and the number of cells in P2 gate is increased;
wherein the P1 gate represents a normal monocyte subpopulation; the P2 gate is a characteristic cell subset of SARS-CoV-2 virus, and the P2 gate subset cells are mononuclear-derived macrophages;
the monocyte-derived macrophages are capable of expressing the cell-specific marker molecules CD11b, CD68, CD80 and CD 163.
2. The application of the angiotensin converting enzyme 2 as a blood mononuclear cell SARS-CoV-2 virus detection target is characterized in that the position change condition of the mononuclear cell expression angiotensin converting enzyme 2 in a blood sample to be detected is investigated through FSC/SSC, and the SARS-CoV-2 virus infection result is judged;
in a blood sample infected by SARS-CoV-2 virus, the angiotensin converting enzyme 2 shows high expression in monocytes in both P1 gate and P2 gate;
wherein the P1 gate represents a normal monocyte subpopulation; the P2 gate is a characteristic cell subset of SARS-CoV-2 virus, and the P2 gate subset cells are mononuclear-derived macrophages; the monocyte-derived macrophages are capable of expressing the cell-specific marker molecules CD11b, CD68, CD80 and CD 163.
3. The use according to claim 2, wherein the human monocyte cell line U937, the human monocyte cell line THP1 and the mouse macrophage cell line RAW264.7 are capable of expressing SARS-CoV-2 virus and the receptor angiotensin converting enzyme 2 of SARS-CoV-2 virus.
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