CN112321700A - Novel coronavirus-infected peripheral blood TCR marker and detection kit and application thereof - Google Patents
Novel coronavirus-infected peripheral blood TCR marker and detection kit and application thereof Download PDFInfo
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Abstract
The invention discloses a novel coronavirus infected peripheral blood TCR marker, a detection kit and application thereof, wherein the marker comprises at least one protein shown in a sequence SpecSeq 1-100. Based on a high-throughput sequencing method, only a small amount of peripheral blood is needed to be taken, RNA is extracted, an immune map library is established through sample processing, and through high-throughput sequencing and TCR data analysis, a characteristic TCR sequence in the peripheral blood infected by the novel coronavirus is firstly determined, and then a test result of a sample to be tested is compared with the characteristic TCR sequence, so that whether the novel coronavirus is infected or not is determined. The invention can simultaneously compare a great number of novel coronavirus infection specific TCR sequences, has higher specificity and accuracy compared with the single detection of one or more markers, and improves the diagnosis efficiency.
Description
Technical Field
The invention belongs to the technical field of genetic engineering, and particularly relates to a novel coronavirus infected peripheral blood TCR marker, a detection kit and application thereof.
Background
The novel coronavirus (SARS-Cov-2) has strong infectivity and great harm, and no specific therapeutic medicine exists at present. The number of diagnosed new coronavirus infection has been over 1 million worldwide, resulting in the death of 50 million people.
The current diagnostic means aiming at the infection of the new coronavirus mainly comprise virus nucleic acid detection, serological IgM and IgG detection and double lung CT. The detection of the new coronavirus nucleic acid mainly depends on PCR amplification reaction, and whether a virus nucleic acid gene exists in a sample to be detected can be detected, so that the detection method is a standard for detecting the new coronavirus. Serological IgM and IgG detection is carried out by detecting a humoral immune response after viral infection. Serological IgM and IgG antibody detection is an auxiliary diagnostic basis for early infection. Positive IgM antibodies indicate that new coronaviruses have been infected. Generally, a human body can generate a virus antigen specific IgM antibody reaction 5 to 7 days after the infection of the new coronavirus, but the IgM antibody reaction is short in maintenance time and disappears quickly. The human body will generate a virus antigen specific IgG antibody response 10-15 days after infection with the new coronavirus. Virus-specific IgG antibodies can be retained in blood for a long time. IgG antibody positive, indicates that a new coronavirus has been infected and some immune resistance has developed. However, not all humans produce IgM or IgG antibodies after infection with the new coronavirus.
B-lymphocytes and T-lymphocytes in the human body are two important types of cells in the adaptive immune system. B cells recognize antigens through BCR on the cell surface, and the BCR is expressed into antibodies and secreted out of the cells in the later period when the B cells are differentiated into plasma cells. T cells recognize antigens via TCRs on the cell surface. The diversity of BCRs and TCRs is the basis for establishing an adaptive immune system. The theoretical value of the diversity of BCR is 1018Theoretical value of TCR diversity is 1014. In various diseases, the diversity or expression level of both BCR and TCR changes with different antigenic stimuli. Therefore, the development of disease can be tracked by using BCR or TCR high-throughput sequencing results. After viral infection in humans, the earliest specific immune response in humans is the T cell response. From the day after viral infection of humans, there is a specific T cell response. The B cell reaction is carried out after the fourth day to generate IgM antibody; IgG antibodies were produced only after the tenth day.
The current diagnostic means aiming at the infection of the new coronavirus mainly comprise virus nucleic acid detection, serological IgM and IgG detection and double lung CT. These three methods all have their own disadvantages. Nucleic acid detection due to sampling limitation, there are 30% -40% false negatives, that is, no virus nucleic acid can be detected. Serological IgM and IgG detection relies on the generation of specific antibody responses in patients following infection with the new coronavirus. Persistent viral antigens are required to stimulate B cells to produce specific antibody responses after infection with new coronavirus. And the double-lung CT detection requires that the patient has lung injury and is diseased.
Disclosure of Invention
The invention aims to: aiming at the defects in the prior art, a novel coronavirus infected peripheral blood TCR marker, a detection kit and application thereof are provided.
The technical scheme adopted by the invention is as follows:
a novel peripheral blood TCR marker of coronavirus infection comprising at least one of the proteins represented by the sequences SpecSeq1-100, as shown in table 1:
TABLE 1 marker sequences
Furthermore, the protein sequence of the marker is that the protein sequence shown by SpecSeq1-100 can express the protein with the same function after one or more bases are substituted, deleted and/or substituted.
Use of the above marker in the preparation of a formulation for detecting or treating a novel coronavirus infection.
Further, the preparation includes a plasmid, a viral vector or a nucleic acid fragment of a T cell receptor containing the marker.
A kit for detecting infection by a novel coronavirus, comprising an antibody capable of specifically binding to the above marker.
An agent comprising an antibody that specifically binds to the marker; the formulations can be used for diagnosis, prognosis, detection or screening of novel coronavirus infections.
A protein chip for detecting the infection of coronavirus is composed of a substrate and the specific antibody spotted on said substrate, which can be specifically combined with said marker.
The principle of the invention is as follows: b-lymphocytes and T-lymphocytes in the human body are two important types of cells in the adaptive immune system. B cells recognize antigens through a B Cell Receptor (BCR) on the cell surface, and later, BCR expresses antibodies and is secreted extracellularly when B cells differentiate into plasma cells. T cells recognize antigens via T Cell Receptors (TCRs) on the cell surface. The diversity of BCRs and TCRs is the basis for establishing an adaptive immune system. BCR varietyTheoretical value of sex is 1018Theoretical value of TCR diversity is 1014. Among the BCR and TCR sequences, epitope 3(CDR3) is the most important part in determining the antigenic specificity, and therefore the sequence of CDR3 is considered to represent the properties of the BCR and TCR sequences.
In various diseases, the diversity or expression level of both BCR and TCR changes with different antigenic stimuli. Therefore, the occurrence and development of diseases can be tracked by using BCR or TCR high-throughput sequencing results. In human cells, after degradation of the senescent protein, fragments thereof are transported to the cell surface and presented to T cells in the immune system by histocompatibility antigen II (MCHII). Antigen fragments presented by normal cells, due to immune tolerance, do not elicit an immune response. Once normal cells become cancerous, the mutated gene expresses an aberrant protein, a fragment of which is presented on the cell surface, which causes a targeted immune response in the human immune system. Therefore, analysis of changes in BCR or TCR enables detection of tumor development and progression.
When the method is applied, a method for analyzing the TCR sequence of the peripheral blood based on high-throughput sequencing is adopted to detect whether the novel coronavirus infection exists, and the specific steps are as follows:
(1) collecting peripheral blood of a subject and a control group, and obtaining the amino acid sequences of the antigenic determinant 3(CDR3) of the TCR of the subject and the control group by high-throughput sequencing to ensure that the total number of the CDR3 sequences of the functional TCR of each sample is not less than 30000 comprehensively;
in the blood collection process, aiming at a new coronavirus infected patient or suspected infected person, peripheral blood is collected and then is directly injected with Trisol lysate into a blood collection purple cap tube by using an injector, cells are lysed, viruses are inactivated, and RNA is extracted.
Specifically, the method comprises the following steps:
collecting peripheral blood: a sterile blood collection tube containing EDTA anticoagulant is used, venous blood is collected by 0.5-1mL (the blood volume just exceeds the bottom hemisphere of the blood collection tube, the blood collection volume is less than 0.5mL or more than 1mL, the follow-up inspection success rate is reduced), and the blood and the anticoagulant are mixed by reversing.
② cell lysis: injecting 5mL of RNAisso plus or TRIzol or SuperfecTRI into the blood sampling tube through the rubber stopper by using a disposable syringe, reversing and mixing, standing at room temperature for 5min to fully crack the cells, and reversing and mixing.
Sample preservation: the lysed peripheral blood samples were stored in a-80 ℃ freezer.
Fourthly, sending the sample: the tubes were wrapped with a foam wrap and sent to the laboratory via a cold chain (possibly transported with dry ice).
(2) The CDR3 sequences of the TCR of each sample were randomly non-back sampled such that the sum of the number of CDR3 sequences of each sample was 30000. For any particular CDR3 sequence X, the number of repeats in the sample is counted as CX；
(3) Analysis of TCR CDR3 data: determination of the sequence of CDR3 of the neocorona-infected TCR marker by analysis:
a) summing and de-duplicating all CDR3 sequences of the control group sample to set as a control sequence set;
b) summarizing and removing the weight of all CDR3 sequences of a new crown patient sample, removing all sequences containing CDR3 sequence repeats in the control sequence set, and setting the sequences as a new crown characteristic sequence set;
c) the CDR3 sequences appearing in two or more samples in the new crown signature sequence set were repeated for "the number of repeats of this sequence in all new crown samples" CXThe sum of (a) x the number of new corona samples containing the sequence "are ordered from high to low, the top 100 being the new corona TCR marker CDR3 sequence.
(4) Judging the risk of infection of the unknown subject with the new corona by using the new corona characteristic index:
a) the index of new crown characteristics was analyzed according to the immune profiles of healthy control group, new crown patients, and subjects with unknown health condition.
Wherein the new crown characteristic index is defined as: in a sample, all CDR3 sequences belonging to the new crown feature sequence set repeat within the sample by the number of times CXThe sum of (a) and (b).
b) When the new coronary signature index of a subject with unknown health condition is higher than the mean value of healthy people and other disease control groups +2 × SD, the person has a higher risk of being infected with new coronary virus; if the new crown characteristic index of the disease is close to the average value of healthy people and other disease control groups, the infection risk is low.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. in the invention, firstly, an artificial intelligence analysis model is established by using TCR high-throughput sequencing data of a control group sample infected by non-novel coronavirus and a new coronary pneumonia (COVID-19) patient, and whether a higher novel coronavirus infection risk exists in a sample to be detected can be clearly judged by comparing with new coronary specificity TCR sequences;
2. the early novel coronavirus infection can be discovered by analyzing TCR change through high-throughput sequencing, and the novel coronavirus infection is identified by utilizing the specific TCR CDR3 sequence of the novel coronarism, so that the reaction of T cells in the immune system of a human to the novel coronarism is analyzed instead of detecting the virus or lung injury, and the method is a novel detection method;
3. in the invention, because a high-throughput sequencing technology is adopted, a great number of specific TCR sequences are compared at the same time, and compared with the method for singly detecting one or more markers, the method has higher specificity and accuracy;
4. the high-throughput sequencing instrument used in the invention has lower cost than large-scale imaging equipment, can be outsourced to a third party, and in addition, the labor cost for sampling and processing is lower than the labor cost for simultaneously detecting various markers and is also lower than the labor cost for a large number of cytological detections, so the detection cost is greatly reduced;
5. according to the invention, only a small amount of peripheral blood is needed to be collected, and various types of samples such as throat swabs and anus swabs are not needed to be collected, so that the sampling is simple, convenient and safe;
6. the TCR CDR3 sequence of the invention can also be used for immunotherapy of neocoronary pneumonia.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 shows the CDR3 sequence of the control group and the new crown signature sequence in the present invention. The abscissa represents the sequence order of the CDR3 sequence of a specific amino acid combination added to the control sequence set or the new crown characteristic sequence set, and the ordinate represents the number of times of the sequence appears repeatedly in a sample CXA logarithmic value of; the immune spectrum of the new corona patient has new corona characteristic sequences with multiple types and high repetition times, few new corona characteristic sequences exist in healthy people, the new corona characteristics of unknown subjects are obvious, and the risk of infecting new corona viruses is high.
FIG. 2 is a graph showing that new crown characteristic indexes of healthy people, other disease control groups and new crown patients are calculated according to a new crown characteristic sequence set, and the healthy people and the other disease control groups have significant differences with the new crown patients, so that the specificity of the new crown characteristic sequence set is proved. Therefore, whether the unknown subject is infected with the new coronavirus can be judged.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
Through immune map analysis, a novel coronavirus infection TCR marker CDR3 sequence set is obtained:
1. sampling and immune profiling
Collecting 1161 control groups (including healthy people and patients not infected with new crowns, 1160 people for establishing a model, 1 healthy person for verification), 81 novel coronavirus infected patients (80 people for establishing a model, 1 person for verification) and peripheral blood (10 mL per person) of 1 unknown health condition subject, obtaining the amino acid sequences of the antigenic determinant 3(CDR3) of the TCR of the subject and the control groups by high-throughput sequencing, and ensuring that the total number of the CDR3 sequences of the functional TCR of each sample is not less than 30000 comprehensively;
2. the CDR3 sequences of the TCR of each sample were randomly non-back sampled such that the sum of the number of CDR3 sequences of each sample was 30000. For any particular CDR3 sequence X, the number of repeats in a single sample sequencing result was counted as CX；
3. By analyzing the TCR CDR3 data, the novel coronavirus infection TCR marker CDR3 sequence was determined:
a) summarizing and de-duplicating all CDR3 sequences of 1160 control group samples for establishing a model, and setting the CDR3 sequences as a control sequence set;
b) all CDR3 sequences of 80 novel coronavirus infection samples used for establishing the model are summed up and de-duplicated, and then all sequences containing sequence repeats in the control sequence set are removed to set the sequences as a novel coronavirus infection characteristic sequence set. As shown in FIG. 1A, the CDR3 sequence with the abscissa representing a specific amino acid combination is added to the sequence of the control sequence set or the sequence set of the novel coronavirus infection characteristic sequence set, and the ordinate represents the repeated occurrence number C of the sequence in a sampleXThe logarithmic value of (c).
c) According to the same mapping method, the immune profiles of 1 healthy person, 1 novel coronavirus-infected patient and 1 subject with unknown health condition are mapped with reference to the control sequence set and the novel coronavirus-infection signature sequence set, as shown in FIGS. 1B-D. As can be seen from the figure, the immune map of the novel coronavirus infected patient contains more types of novel coronavirus infection characteristic sequences with higher repeated occurrence frequency; in the immune map of a healthy person, only a few novel coronavirus infection characteristic sequences exist; the unknown healthy subjects have the characteristic sequences of the novel coronavirus infection higher than those of healthy persons, which indicates that the persons are at higher risk of suffering the novel coronavirus infection.
d) Concentrating the infection characteristic sequence of the novel coronavirus, and determining the repeated occurrence times C of the CDR3 sequences in the single sample of the sequence in the sampleXThe total sum of (a) x the number of samples participating in modeling of the novel coronavirus infection containing the sequence is ranked from high to low, and the top 100 are the sequences of the CDR3 marker of the novel coronavirus infection TCR.
Example 2
The specificity of the CDR3 sequence set of the novel coronavirus infection TCR marker is verified:
1. sampling and immune profiling
Collecting peripheral blood (10 mL per person) of 224 tumor patients without new crown infection and 5 subjects with unknown health conditions, and obtaining the amino acid sequences of antigenic determinant 3(CDR3) of TCR of the subjects and the control group by high-throughput sequencing to ensure that the total number of CDR3 sequences of functional TCR of each sample is not less than 30000 comprehensively; the CDR3 sequences of the TCR of each sample were randomly non-back sampled such that the sum of the number of CDR3 sequences of each sample was 30000.
2. 100 healthy and 45 patients with non-tumor diseases were randomly selected from the control group of example 1.
3. The index characteristic of neocoronary infection was analyzed on the basis of the immune profiles of 100 healthy persons from example 1, 45 non-neoplastic patients not infected with neocoronary, 80 neocoronary infected persons, and 224 newly acquired neoplastic patients not infected with neocoronary, 5 subjects of unknown health status from example 2.
Wherein the index characteristic of neocoronal infection is defined as: in a sample, all CDR3 sequences belonging to the set of signature sequences for new crown infections are repeated within the sample for a number of times CXThe sum of (a) and (b).
The results of the analysis are shown in Table 2 below and FIG. 2. The specificity of the characteristic sequence set of the new crown infection is proved by the remarkable differences of the new crown infection group and healthy people (p is 1.4E-117), non-tumor diseases (p is 5.6E-71) and tumors (p is 4.7E-216).
TABLE 2 novel coronavirus infection characteristic indices for different sample groups
4. Analyzing the novel characteristic indexes of coronavirus infection in each group (table 3 below), wherein the characteristic index of new coronary infection in the first 3 people in the unknown health condition subjects (test samples) is higher than the average value of healthy people and other disease control groups, namely +2 × SD (347+2 × 1048 ═ 2443), and the 3 people are at higher risk of new coronary infection; the index of the new crown infection characteristic of the later 2 people is close to the average value of healthy people or other disease groups, and the risk of new crown infection is low. After comparison with the clinical examination results, the first 3 patients were confirmed to be new crown infected patients, and the second 2 were healthy people. This example demonstrates the feasibility of predicting a subject's risk of contracting a new corona infection using a set of signature sequences characteristic of a new corona infection and an index characteristic of a new corona infection.
TABLE 3 analysis table of characteristic indexes of novel coronavirus infection
In conclusion, the CDR3 sequence of the novel coronavirus infection TCR marker of the present invention has significant specificity for novel coronavirus infection, and can be used for predicting the risk of a subject suffering from novel coronavirus infection, and for biological immunotherapy of novel coronavirus infection in the future.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Sequence listing
<110> Hospital for people in Sichuan province
THE FIFTH MEDICAL CENTER OF THE CHINESE PEOPLE'S LIBERATION ARMY GENERAL Hospital
The First Affiliated Hospital of Zhengzhou University
<120> novel coronavirus infected peripheral blood TCR marker and detection kit and application thereof
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Ala Thr Ser Ser Gly Asn Ile Asn Glu Gln Phe
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Ala Ser Ser Ser Gln Gln Leu Val Asp Arg Leu Trp Gly Gln Tyr
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Ser Ala Arg Asp Asn Asn Gly Val Gly Leu Ala Gly Leu Tyr Glu Gln
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Ala Ser Ser Thr Ala Tyr Arg Asp Asn Ser Pro Leu His
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Ala Thr Leu Thr Gly Gly Leu Lys Thr Gln Tyr
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Ala Thr Glu Met Gly Gln Arg Gly Thr Tyr Asn Glu Gln Phe
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Ala Ser Ser Gln Asp Glu Val Arg Val Ser Tyr Asn Glu Gln Phe
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Ala Ser Pro Ile Ala Gly Pro Tyr Glu Gln Tyr
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Ala Ser Ser Ser Gly Trp Pro Gln Glu Thr Gln Tyr
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Ala Ser Ser Lys Arg Ala Tyr Pro Tyr Asn Glu Gln Phe
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Ala Ser Ser Tyr Thr Ala Asp Ser Ser Asp Thr Gln Tyr
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<210> 27
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Ala Ser Ser Leu Asp Val Ala Ser Gly Tyr Thr Asp Thr Gln Tyr
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Ser Val Leu Thr Gly Arg Glu Thr Asp Thr Gln Tyr
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Ala Ser Ser Pro Val Arg Ala Asp Gly Asn Thr Ile Tyr
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Ala Ser Ser Phe Ser Asn Phe Trp Thr Asp Thr Gln Tyr
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Ala Ser Arg Ser Glu Arg Gly Glu Ala Phe
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Ala Ser Ser Leu Gln Gly Ala Ser Val Asn Gln Pro Gln His
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Ala Thr Ser Gly Leu Pro His Thr Asp Thr Gln Tyr
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Ala Ser Ser Asp Trp Leu Gly Glu Leu Phe
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Ala Ser Ser Gln Val Trp Asp Lys Thr Tyr Glu Gln Tyr
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Ala Ser Gln Gly Ser Leu Asp Ser Pro Leu His
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<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ile Val Met Phe Gly Glu Gly Arg Asn Thr Glu Ala Phe
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<212> PRT
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Ala Ser Arg Ala Gly Ile Ile Ser Gly Asn Thr Ile Tyr
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<210> 39
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Lys Val Thr Gly Thr Tyr Glu Gln Tyr
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<210> 40
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<212> PRT
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Ala Thr Glu Val Asp Ser Thr Asn Glu Lys Leu Phe
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<210> 41
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<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ser Leu Leu Gly Gln Arg Glu Gln Tyr
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Ala Ser Ser Ala Thr Ser Gln Asp Gln Tyr
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Ala Ser Arg Gly Thr Gly Asp Met Ser Asn Gln Pro Gln His
1 5 10
<210> 44
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Thr Pro Ser Ser Asn Gln Tyr Asn Glu Gln Phe
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<210> 45
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 45
Ala Ser Ser Leu Arg Leu Ala Asp Met Thr Asp Thr Gln Tyr
1 5 10
<210> 46
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ser Gln Ser Asp Gly Asp Pro His Tyr Glu Gln Tyr
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<210> 47
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<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ser Leu Glu Trp Thr Ser Val Asp Thr Gln Tyr
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<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ser Phe Gly Thr Ser Gly Asn Leu Phe
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Ala Ser Cys Thr Ser Gly Ser Leu Ser His Glu Gln Phe
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Ala Ser Ser Tyr Pro Asp Val His Glu Gln Phe
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<210> 51
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<212> PRT
<213> Artificial Sequence (Artificial Sequence)
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Ala Ser Ser Gln Ala Pro Thr Ser Ser Tyr Asn Glu Gln Phe
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<210> 52
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<212> PRT
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Ala Ser Ser Lys Val Arg Gly Asn Tyr Asn Glu Gln Phe
1 5 10
<210> 53
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 53
Ala Ser Ser Leu Val Trp Leu Gly Lys Trp Asn Glu Gln Phe
1 5 10
<210> 54
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 54
Ala Ile Ser Thr Asp Trp Thr Ser His Phe Ser Tyr Glu Gln Tyr
1 5 10 15
<210> 55
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 55
Arg Pro Gly Glu Arg Gly Lys Gln Phe
1 5
<210> 56
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 56
Ala Thr Ser Asp Ser Glu Asp Arg Thr Gln Tyr
1 5 10
<210> 57
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 57
Ala Ser Ser Tyr Leu Phe Pro Gly Leu Ala Glu Asp Glu Gln Tyr
1 5 10 15
<210> 58
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 58
Ala Thr Ser Asp Leu Gly Gly Arg Gly Glu Gln Phe
1 5 10
<210> 59
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 59
Ala Ser Ser Ser Gly Gln Gly Ser Ile Tyr Asn Glu Gln Phe
1 5 10
<210> 60
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 60
Ala Ser Ser Val Thr Glu Phe Thr Tyr Gly Tyr Thr
1 5 10
<210> 61
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 61
Ala Ser Ser Tyr Met Gly Gln Val Ser Ser Tyr Glu Gln Tyr
1 5 10
<210> 62
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 62
Ser Val Glu Arg Asp Ser Ile Asn Thr Glu Ala Phe
1 5 10
<210> 63
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 63
Ala Ser Asp Gly Arg Thr Glu Ala Phe
1 5
<210> 64
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 64
Ala Thr Thr Pro Arg Pro Asn Thr Gly Glu Leu Phe
1 5 10
<210> 65
<211> 10
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 65
Ala Ser Ser Trp Phe Glu Asn Glu Gln Phe
1 5 10
<210> 66
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 66
Ala Ser Ser Lys Gly Arg Tyr Asn Thr Glu Ala Phe
1 5 10
<210> 67
<211> 13
<212> PRT
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<400> 67
Gln Gln Leu Leu Arg Val Gly Ala Arg Gln Arg Ala Val
1 5 10
<210> 68
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 68
Ala Ser Ser Tyr Ser Val Gly Leu Glu Val Tyr Glu Gln Tyr
1 5 10
<210> 69
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 69
Ala Ser Ser Tyr Thr Pro Asp Arg Ala Asn Glu Ala Phe
1 5 10
<210> 70
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 70
Ala Thr Ser Asp Leu Val Ser Gly Glu Asp Glu Gln Phe
1 5 10
<210> 71
<211> 16
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 71
Ala Ser Ser Pro Asp Val Gly Gly Gln Val Val Asn Tyr Gly Tyr Thr
1 5 10 15
<210> 72
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 72
Ala Ser Ser Pro Met Thr Gly Gln Gln Asp Thr Gln Tyr
1 5 10
<210> 73
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 73
Ala Thr Ala Gly Gly Arg Thr Ser Ser Tyr Asn Glu Gln Phe
1 5 10
<210> 74
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 74
Ala Ser Ser Pro Leu Ala Gly Asp Pro Gln His
1 5 10
<210> 75
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 75
Ala Ser Arg Tyr Gly Thr Ser Leu Ile Asn Glu Gln Phe
1 5 10
<210> 76
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 76
Ala Ser Ser Ser Thr Arg Thr Gly Glu Lys Leu Phe
1 5 10
<210> 77
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 77
Ala Ser Ser Leu Gly Ile Gln Gly Pro Glu Gln Tyr
1 5 10
<210> 78
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 78
Ala Ser Asn Ser Pro Gly Gln Gly Ala Tyr Glu Gln Tyr
1 5 10
<210> 79
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 79
Ala Ser Ser Gln Glu Asn Ser Gly Arg Thr Ser Gly Ala Asn Val Leu
1 5 10 15
Thr
<210> 80
<211> 18
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 80
Ala Thr Ser Asp Leu His Trp Ile Arg Glu Leu Ile Ala Lys Asn Ile
1 5 10 15
Gln Tyr
<210> 81
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 81
Ala Ser Ser Leu Glu Ser Arg Pro Gly Tyr Thr
1 5 10
<210> 82
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 82
Ser Ala Arg Asp Leu Gly Thr Gly Asn Glu Gln Phe
1 5 10
<210> 83
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 83
Ala Ile Arg Ala Thr Ser Gly Gly Thr Asp Thr Gln Tyr
1 5 10
<210> 84
<211> 17
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 84
Ala Ser Ser Gln Glu Gly Ser Gly Arg Val Thr Ala Lys Asn Ile Gln
1 5 10 15
Tyr
<210> 85
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 85
Ala Ser Ser Pro Asn Glu His Ser Asn Gln Pro Gln His
1 5 10
<210> 86
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 86
Ala Leu Arg Asp Lys Gly Phe Thr Glu Ala Phe
1 5 10
<210> 87
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 87
Ser Val Glu Asp Arg Glu Ala Tyr Gly Tyr Thr
1 5 10
<210> 88
<211> 9
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 88
Ala Thr Ser Trp Asp Gly Glu Ala Phe
1 5
<210> 89
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 89
Ser Ala Arg Asp Arg Ser Ser Asp Tyr Ser Tyr Glu Gln Tyr
1 5 10
<210> 90
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 90
Ala Ser Ser Thr Lys Arg Arg Pro Pro Tyr Asn Glu Gln Phe
1 5 10
<210> 91
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 91
Ala Ser Leu Gly Tyr Ser Gly Asn Thr Ile Tyr
1 5 10
<210> 92
<211> 15
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 92
Ala Ser Ser Gln Asp Ser Glu Arg Glu Tyr Tyr Asn Glu Gln Phe
1 5 10 15
<210> 93
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 93
Ala Ser Ser Gln Asp Ala Met Asp Ala Asp Thr Gln Tyr
1 5 10
<210> 94
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 94
Ala Ser Ser Asp Ser Arg Arg Thr Asn Tyr Gly Tyr Thr
1 5 10
<210> 95
<211> 14
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 95
Ala Ser Ser Leu Asn Leu Leu Gly Lys Ala Gly Glu Leu Phe
1 5 10
<210> 96
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 96
Ser Ala Ala Arg Asp Ser Tyr Ser Ser Tyr Glu Gln Tyr
1 5 10
<210> 97
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 97
Ala Thr Ser Asp Leu Gly Ser Gly His Glu Gln Phe
1 5 10
<210> 98
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 98
Ala Thr Arg Glu Glu Leu Pro Thr Tyr Asn Glu Gln Phe
1 5 10
<210> 99
<211> 13
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 99
Ala Ser Ser Gln Asp Gln Ala Gly Tyr Glu Lys Leu Phe
1 5 10
<210> 100
<211> 12
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 100
Ala Ser Arg Lys Gly Gln Glu Gln Glu Thr Gln Tyr
1 5 10
Claims (7)
1. A novel peripheral blood TCR marker of coronavirus infection comprising at least one protein of sequence SpecSeq 1-100.
2. The novel peripheral blood TCR marker for coronavirus infection according to claim 1, wherein the protein sequence of the marker is a protein capable of expressing the same function after substitution, deletion and/or substitution of one or more bases of the sequence shown in SpecSeq 1-100.
3. Use of a marker according to claim 1 or 2 for the preparation of a formulation for the detection or treatment of a novel coronavirus infection.
4. The use of claim 3, wherein the preparation comprises a plasmid, viral vector or nucleic acid fragment of a T cell receptor comprising the marker.
5. A kit for the detection of a novel coronavirus infection, comprising an antibody that specifically binds to the marker of claim 1.
6. An agent comprising an antibody that specifically binds to the marker of claim 1.
7. A protein chip for detecting infection by a novel coronavirus, said protein chip comprising a substrate and a specific antibody spotted on the substrate, wherein said specific antibody is an antibody capable of specifically binding to the marker of claim 1.
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| CN202010995218.0A CN112321700A (en) | 2020-09-21 | 2020-09-21 | Novel coronavirus-infected peripheral blood TCR marker and detection kit and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010995218.0A CN112321700A (en) | 2020-09-21 | 2020-09-21 | Novel coronavirus-infected peripheral blood TCR marker and detection kit and application thereof |
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| Publication Number | Publication Date |
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| CN112321700A true CN112321700A (en) | 2021-02-05 |
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| CN202010995218.0A Pending CN112321700A (en) | 2020-09-21 | 2020-09-21 | Novel coronavirus-infected peripheral blood TCR marker and detection kit and application thereof |
Country Status (1)
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| CN (1) | CN112321700A (en) |
Cited By (3)
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| CN113030473A (en) * | 2021-03-15 | 2021-06-25 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute B lymphocyte leukemia and detection kit and application thereof |
| CN113150113A (en) * | 2020-05-23 | 2021-07-23 | 湖南源品细胞生物科技有限公司 | TCR enrichment clone type and acquisition method and application thereof |
| CN113567682A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of Alzheimer disease and detection kit and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113030473A (en) * | 2021-03-15 | 2021-06-25 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of acute B lymphocyte leukemia and detection kit and application thereof |
| WO2022194039A1 (en) * | 2021-03-15 | 2022-09-22 | 成都益安博生物技术有限公司 | Peripheral blood tcr marker for acute b lymphocytic leukemia, and detection kit thereof and use thereof |
| CN113567682A (en) * | 2021-07-23 | 2021-10-29 | 成都益安博生物技术有限公司 | Peripheral blood TCR marker of Alzheimer disease and detection kit and application thereof |
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