阅读说明：本技术 免疫球蛋白的检测试剂在制备结直肠癌诊断剂的应用 (Application of immunoglobulin detection reagent in preparation of colorectal cancer diagnostic agent ) 是由 杨湘玲 刘瑞贤 王学钦 张瀞丹 赵璐 徐重 刘焕亮 于 2019-10-12 设计创作，主要内容包括：本发明属于生物医药领域,涉及免疫球蛋白检测试剂在制备结直肠癌诊断或预后试剂/试剂盒中的应用。本发明研究发现,特定免疫球蛋白在结肠直肠癌中差异表达,且具有很大的差异性和可信度。将单一免疫球蛋白及不同免疫球蛋白组合作为结直肠癌诊断生物标记物,有利于提高结肠直肠癌的诊疗水平,有效防治结直肠癌。(The invention belongs to the field of biomedicine, and relates to an application of an immunoglobulin detection reagent in preparation of a colorectal cancer diagnosis or prognosis reagent/kit. The research of the invention finds that the specific immunoglobulin is differentially expressed in colorectal cancer and has great difference and reliability. The single immunoglobulin and different immunoglobulins are combined to be used as the colorectal cancer diagnosis biomarker, so that the diagnosis and treatment level of colorectal cancer is improved, and the colorectal cancer is effectively prevented and treated.)

1. Use of a detection reagent for immunoglobulins for the preparation of a colorectal cancer diagnostic reagent/kit, characterized in that the immunoglobulins comprise IgD and/or IgE.

2. The use according to claim 1, wherein the immunoglobulin comprises IgD and IgE;

preferably, the immunoglobulin is IgD and IgE;

preferably, the immunoglobulin further comprises one or more of IgG1, IgG3, or IgA 1;

preferably, the immunoglobulin is IgD, IgE and IgG 3;

preferably, the immunoglobulin is IgD, IgE, IgG1 and IgG 3;

preferably, the immunoglobulin is IgD, IgE, IgG3 and IgA 1.

3. The use of claim 1, wherein the detection reagent comprises detecting the amount of expression of mRNA of immunoglobulin; or a reagent for detecting the expression level of the immunoglobulin or for detecting the biological activity of the immunoglobulin; preferably, the detection reagent comprises a reagent for detecting the expression amount of immunoglobulin; preferably, the detection sample of the detection reagent is blood; more preferably plasma.

4. The use of claim 1, wherein the detection reagent comprises an antibody, antibody functional fragment or conjugated antibody per immunoglobulin; preferably, the conjugated antibody is a fluorescein conjugated antibody or a biological enzyme conjugated antibody.

5. The use of claim 1, wherein the kit is an ELISA kit.

6. The use of claim 1, wherein the detection method of the detection reagent comprises any one or more of an ELISA method, a protein chip method, a liquid chromatography method and a flow cytometry method; preferably, it is a protein chip method or an ELISA method.

7. The use of claim 1, wherein when the immunoglobulin is IgD, the detection reagent determines that the critical value of the IgD protein expression level for high risk or low risk of colorectal cancer is in the range of 1906.41pg/ml to 6168.47 pg/ml;

preferably, the critical value of the IgD protein expression amount is 5842.19 pg/ml; when the expression quantity of the IgD protein is less than or equal to 5842.19pg/ml, the colorectal cancer risk is low; the detection result of the detection reagent is as follows: if the expression amount of the IgD protein is greater than 5842.19pg/ml, the colorectal cancer is at high risk.

8. The use of claim 1, wherein when the immunoglobulin is IgE, the detection reagent determines that the critical value of the expression level of IgE protein at high risk or low risk of colorectal cancer is 978.68pg/ml to 1305.34 pg/ml;

preferably, the critical value of the IgE protein expression amount is 991.94 pg/ml; when the expression quantity of the IgE protein is less than or equal to 991.94pg/ml, the colorectal cancer is at high risk; the detection result of the detection reagent is as follows: the expression level of IgE protein is greater than 991.94pg/ml, and the colorectal cancer risk is low.

9. The use according to claim 2, wherein the detection reagent determines the critical value of the risk index X for high or low risk of colorectal cancer in the range of 0.5000 to 0.7416 when the number of immunoglobulins is at least two;

preferably, when the immunoglobulins are IgD and IgE, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.00158C)_IgE+0.000131C_IgD+1.474))) of said C_IgEIs the concentration of IgE, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5920-0.6418; preferably, the critical value of the risk index X is 0.6039;

preferably, when the immunoglobulins are IgD, IgE and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001602C)_IgE+0.000119C_IgG3+0.0001C_IgD+1.091))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5000-0.6762; preferably, the critical value of the risk index X is 0.5265;

preferably, when the immunoglobulins are IgD, IgE, IgG1 and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001904C)_IgE+0.000273C_IgG3+0.000107C_IgD-0.000008C_IgG1+2.047))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgG1The concentration of IgG1 is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.6231-0.6659; preferably, the critical value of the risk index X is 0.6280;

preferably, when the immunoglobulins are IgD, IgE, IgG3 and IgA1, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001587C)_IgE+0.000212C_IgG3+0.000110C_IgD-0.000019C_IgA1+2.769))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgA1The concentration of IgA1 is the concentration of a detection sample diluted by 10000 times, and the critical value range of the risk index X is 0.5933-0.7416; preferably, the critical value of the risk index X is 0.7374;

preferably, the detection result of the detection reagent is as follows: when the risk index X is less than or equal to a critical value, the colorectal cancer is at low risk; when the risk index X > cutoff value, there is a high risk of colorectal cancer.

10. A diagnostic system for colorectal cancer, wherein the detection system comprises:

a) a detection means: the detection component is used for detecting the expression amount of immunoglobulin of a diagnosis object, and the immunoglobulin comprises IgD and/or IgE;

preferably, the immunoglobulin is IgD and IgE;

preferably, the immunoglobulin further comprises one or more of IgG1, IgG3, or IgA 1;

preferably, the immunoglobulin is IgD, IgE and IgG 3;

preferably, the immunoglobulin is IgD, IgE, IgG1 and IgG 3;

preferably, the immunoglobulin is IgD, IgE, IgG3 and IgA 1;

b) a result judgment means: the result judging component is used for obtaining the possibility or the risk value of the colorectal cancer according to the expression quantity of the immunoglobulin detected by the detecting component;

preferably, the detection component comprises one or more of a microplate reader, a laser scanner, a flow cytometer and a liquid chromatograph; more preferably, the detection component is one or two of a laser scanner and a microplate reader;

preferably, the result judging means comprises software including an input module, an analysis module and an output module; the input module is used for inputting the expression quantity of the immunoglobulin; the analysis module is used for analyzing the colorectal cancer morbidity possibility or risk value according to the expression quantity of the immunoglobulin; the output module is used for outputting the analysis result of the analysis module;

preferably, the expression level of the immunoglobulin is the expression level of mRNA of the immunoglobulin; or the amount of protein expressed; more preferably the amount of protein expressed;

preferably, the diagnostic sample of the diagnostic system is a blood sample; more preferably a plasma sample;

preferably, when the immunoglobulin is IgD, the value of the critical value of the IgD protein expression quantity for judging the high risk or low risk of colorectal cancer by the detection reagent is in the range of 1906.41pg/ml to 6168.47 pg/ml;

preferably, the critical value of the IgD protein expression amount is 5842.19 pg/ml; when the expression quantity of the IgD protein is less than or equal to 5842.19pg/ml, the colorectal cancer risk is low; the detection result of the detection reagent is as follows: if the expression quantity of the IgD protein is greater than 5842.19pg/ml, the colorectal cancer is high in risk;

preferably, when the immunoglobulin is IgE, the value of the critical value of the IgE protein expression quantity for judging the high risk or the low risk of the colorectal cancer by the detection reagent is in the range of 978.68pg/ml to 1305.34 pg/ml;

preferably, the critical value of the IgE protein expression amount is 991.94 pg/ml; when the expression quantity of the IgE protein is less than or equal to 991.94pg/ml, the colorectal cancer is at high risk; the detection result of the detection reagent is as follows: the expression amount of the IgE protein is greater than 991.94pg/ml, and the colorectal cancer risk is low;

preferably, when the number of the immunoglobulins is at least two, the critical value of the risk index X for judging the high risk or the low risk of colorectal cancer by the detection reagent is in the range of 0.5000-0.7416;

preferably, when the immunoglobulins are IgD and IgE, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.00158C)_IgE+0.000131C_IgD+1.474))) of said C_IgEIs the concentration of IgE, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5920-0.6418; preferably, the critical value of the risk index X is 0.6039;

preferably, when the immunoglobulins are IgD, IgE and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001602C)_IgE+0.000119C_IgG3+0.0001C_IgD+1.091))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5000-0.6762; preferably, the critical value of the risk index X is 0.5265;

preferably, when the immunoglobulins are IgD, IgE, IgG1 and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001904C)_IgE+0.000273C_IgG3+0.000107C_IgD-0.000008C_IgG1+2.047))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgG1The concentration of IgG1 is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.6231-0.6659; preferably, the critical value of the risk index X is 0.6280;

preferably, when the immunoglobulins are IgD, IgE, IgG3 and IgA1, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001587C)_IgE+0.000212C_IgG3+0.000110C_IgD-0.000019C_IgA1+2.769))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgA1The concentration of IgA1 is the concentration of a detection sample diluted by 10000 times, and the critical value range of the risk index X is 0.5933-0.7416; preferably, the critical value of the risk index X is 0.7374;

preferably, the detection result of the detection reagent is as follows: when the risk index X is less than or equal to a critical value, the colorectal cancer is at low risk; when the risk index X > cutoff value, there is a high risk of colorectal cancer.

Technical Field

The invention belongs to the field of biomedicine, and relates to application of a detection reagent of immunoglobulin in preparation of a colorectal cancer diagnosis reagent/kit and a colorectal cancer diagnosis reagent/kit.

Background

Colorectal cancer (CRC) is the most common malignancy of the digestive tract, the second largest cancer in the world, with high morbidity and mortality at the third and fourth sites, respectively. In recent years, with the improvement of living standard of people in China, the change of living style and the aging of population, the incidence of colorectal cancer in China shows a trend of gradually rising, and the third incidence and the fifth incidence of malignant tumor in China seriously threaten the health and life of people. Early discovery, early diagnosis and early treatment are the key points for treating colorectal cancer. However, most colorectal cancers have no obvious symptoms at the initial stage, and are diagnosed by enteritis as the disease progresses, so that most colorectal cancer patients are in the tumor progression stage when diagnosed, the optimal treatment time is lost, the five-year survival rate is less than 20%, and the prognosis is poor. Therefore, exploring an early, rapid and large-scale diagnosis method for colorectal cancer and improving the diagnosis and treatment level are problems to be solved urgently for preventing and treating colorectal cancer at present.

Human serum contains a large amount of Immunoglobulin (Ig), a large Y-shaped protein that is secreted mainly by plasma cells and used by the immune system to identify and neutralize foreign substances such as pathogens like bacteria, viruses, etc., and has various Immunoglobulin subtypes; more importantly, different subtypes of immunoglobulins fluctuate when the body is infected; compared with other protein molecules, the immunoglobulin has more stable property and longer half-life, and the characteristics ensure that the expression levels of different immunoglobulin subtypes in blood plasma have wide prospects in the diagnosis and application of diseases such as infection, tumor and the like.

Igs can be classified into five classes, i.e., IgA, IgG, IgM, IgD, IgE, depending on the number of Y-shaped structures and the type of heavy chain, which are γ, μ, α, δ, and ε, respectively. Igs vary in biological properties, functional regions, and the ability to bind different antigens. Among them, IgA is a highly glycosylated protein, and exists mainly in two forms, IgA1 and IgA 2. IgG is the highest content of immunoglobulin in serum, accounts for 70-75%, and is also the most important anti-infective molecule. IgG is classified into 4 subtypes, IgG1, IgG2, IgG3 and IgG4, respectively, according to the difference in antigenicity of its molecular backbone (γ chain) and the difference in the number and position of disulfide bonds. IgM, IgD and IgE have only one form.

Each Ig subtype has different functions, and the content of the Ig is too low or too high or deficient, which causes corresponding immunologic hypofunction and causes corresponding diseases. If only the total content of one Ig is detected, and the content of each subtype is not detected, misdiagnosis is often caused. Because many patients have normal total levels of an Ig but are deficient or contain abnormal levels of individual subtypes. If only antibiotic therapy is given, but not immunotherapy is given in time, unnecessary troubles are caused to the patient. Therefore, detection of each Ig subtype is of great clinical significance.

Disclosure of Invention

The invention aims to provide a molecular marker of colorectal cancer, a diagnostic reagent of the molecular marker and application of the diagnostic reagent.

The above object of the present invention is achieved by the following technical means:

in one aspect, the present invention provides the use of a detection reagent for immunoglobulins comprising IgD and/or IgE in the preparation of a colorectal cancer diagnostic reagent/kit.

As a preferred embodiment, the immunoglobulin comprises IgD and IgE.

As a preferred embodiment, the immunoglobulin is IgD and IgE.

As a preferred embodiment, the immunoglobulin further comprises one or more of IgG1, IgG3, or IgA 1;

as a preferred embodiment, the immunoglobulin is IgD, IgE and IgG 3.

As a preferred embodiment, the immunoglobulin is IgD, IgE, IgG1 and IgG 3.

As a preferred embodiment, the immunoglobulin is IgD, IgE, IgG3 and IgA 1.

The inventor collects blood of healthy people (preferably plasma samples) and plasma samples of colorectal cancer patients for detection, obtains the standardized abundance of different plasma proinflammatory/anti-inflammatory factors through data processing and analysis, and discovers that the differential expression of immunoglobulin subtypes, namely different immunoglobulins, can be used as biomarkers for colorectal cancer diagnosis.

In addition, when the immunoglobulin is one type, the detection reagent detects the immunoglobulin; when the number of the immunoglobulin is at least two, the detection reagent detects each immunoglobulin.

The reagent for detecting an immunoglobulin comprises a step of detecting the expression level of mRNA of the immunoglobulin; or detecting the expression level of the immunoglobulin, or detecting one or more of the biological activities of the immunoglobulin. In a preferred embodiment of the present invention, the reagent for detecting immunoglobulin comprises detecting the expression level of immunoglobulin.

In a preferred embodiment, the expression level of the immunoglobulin is the concentration of the immunoglobulin in the test sample; in a more preferred embodiment, the concentration of immunoglobulin in plasma is expressed.

In a preferred embodiment, the detection reagent comprises an antibody, functional fragment of an antibody, or conjugated antibody that detects each immunoglobulin.

The antibody may be a monoclonal antibody, a polyclonal antibody, a multivalent antibody, a multispecific antibody (e.g., bispecific antibody), and/or an antibody fragment linked to a proteasome. The antibody may be a chimeric antibody, a humanized antibody, a CDR-grafted antibody or a human-type antibody. Antibody fragments may be, for example, Fab, Fab ', F (ab') 2, Fv, Fd, single chain Fv (scFv), Fv with disulfide bonds (sdFv), or VL, VH domains. The antibody may be in a conjugated form, e.g., conjugated to a label, a detectable label, or a cytotoxic agent.

In a preferred embodiment, the conjugated antibody is a fluorescein conjugated antibody or a biological enzyme conjugated antibody.

In a preferred embodiment, the kit is an ELISA kit, and further, an ELISA kit based on an anti-human immunoglobulin conjugated antibody, and the detection kit is used for detecting the expression level of immunoglobulin.

The detection method of the detection reagent comprises any one or more of an ELISA method, a protein chip method, a liquid chromatography method, an immunoturbidimetry method and a flow cytometry method; in a preferred embodiment, the detection method of the detection reagent is one or more of a protein chip method, an ELISA method and an immunoturbidimetry method; in a more preferred embodiment, the detection method of the detection reagent is a protein chip method or an ELISA method.

In a preferred embodiment, when the immunoglobulin is IgD, the value of the critical value of the IgD protein expression level for determining the high risk or low risk of colorectal cancer by the detection reagent is in the range of 1906.41pg/ml to 6168.47 pg/ml.

In a preferred embodiment, the critical value of the expression level of the IgD protein is 5842.19 pg/ml; when the expression quantity of the IgD protein is less than or equal to 5842.19pg/ml, the colorectal cancer risk is low; the detection result of the detection reagent is as follows: if the expression amount of the IgD protein is greater than 5842.19pg/ml, the colorectal cancer is at high risk.

In a preferred embodiment, when the immunoglobulin is IgE, the detection reagent determines that the critical value of the expression level of the IgE protein with high risk or low risk of colorectal cancer is 978.68pg/ml to 1305.34 pg/ml.

In a preferred embodiment, the critical value of the IgE protein expression amount is 991.94 pg/ml; when the expression quantity of the IgE protein is less than or equal to 991.94pg/ml, the colorectal cancer is at high risk; the detection result of the detection reagent is as follows: the expression level of IgE protein is greater than 991.94pg/ml, and the colorectal cancer risk is low.

As an exemplary embodiment of the present invention, a standard curve method may be used to calculate the expression levels of different immunoglobulins.

In a preferred embodiment, when there are at least two immunoglobulins, the threshold value of the risk index X for determining a high risk or a low risk of colorectal cancer is in the range of 0.5000 to 0.7416.

As a preferred embodiment, when the immunoglobulins are IgD and IgE, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.00158C)_IgE+0.000131C_IgD+1.474))) of said C_IgEIs the concentration of IgE, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5920-0.6418; in a preferred embodiment, the critical value of the risk index X is 0.6039.

As a preferred embodiment, when the immunoglobulins are IgD, IgE and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001602C)_IgE+0.000119C_IgG3+0.0001C_IgD+1.091))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5000-0.6762; in a preferred embodiment, the critical value of the risk index X is 0.5265.

As a preferred embodiment, when the immunoglobulins are IgD, IgE, IgG1 and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001904C)_IgE+0.000273C_IgG3+0.000107C_IgD-0.000008C_IgG1+2.047))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgG1The concentration of IgG1 is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.6231-0.6659; in a preferred embodiment, the critical value of the risk index X is 0.6280.

As a preferred embodiment, when the immunoglobulins are IgD, IgE, IgG3 and IgA1, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001587C)_IgE+0.000212C_IgG3+0.000110C_IgD-0.000019C_IgA1+2.769))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgA1The concentration of IgA1 is the concentration of a detection sample diluted by 10000 times, and the critical value range of the risk index X is 0.5933-0.7416; in a preferred embodiment, the critical value of the risk index X is 0.7374.

In a preferred embodiment, the detection result of the detection reagent is: when the risk index X is less than or equal to a critical value, the colorectal cancer is at low risk; when the risk index X > cutoff value, there is a high risk of colorectal cancer.

In a preferred embodiment, the sample to be tested by the test reagent is blood; in a preferred embodiment, the sample to be detected by the detection reagent is plasma.

In another aspect, the present invention provides a colorectal cancer diagnosis system, the detection system comprising:

a) a detection means: the detection means is used for detecting the expression level of the immunoglobulin of the diagnosis object;

as a preferred embodiment, the immunoglobulin comprises IgD and/or IgE;

as a preferred embodiment, the immunoglobulin is IgD and IgE;

as a preferred embodiment, the immunoglobulin further comprises one or more of IgG1, IgG3, or IgA 1;

as a preferred embodiment, the immunoglobulin is IgD, IgE and IgG 3;

as a preferred embodiment, the immunoglobulin is IgD, IgE, IgG1 and IgG 3;

as a preferred embodiment, the immunoglobulin is IgD, IgE, IgG3 and IgA 1.

b) A result judgment means: the result judging component is used for obtaining the possibility or the risk value of the colorectal cancer according to the expression quantity of the immunoglobulin detected by the detecting component.

In a preferred embodiment, the detection component is one or more of a microplate reader, a laser scanner, a flow cytometer and a liquid chromatograph; in a preferred embodiment, the detection member is one or two of a microplate reader and a laser scanner.

In a preferred embodiment, the result judging means is software, which comprises an input module, an analysis module and an output module; the input module is used for inputting the expression quantity of the immunoglobulin; the analysis module is used for analyzing the colorectal cancer morbidity possibility or risk value according to the expression quantity of the immunoglobulin; the output module is used for outputting the analysis result of the analysis module.

Wherein in the system, the expression level of the immunoglobulin is the expression level of mRNA of the immunoglobulin; or the expression level of the protein. In a preferred embodiment, the detection means detects the expression level of immunoglobulin.

Wherein the diagnostic sample of the diagnostic system is blood; more preferably plasma.

In a preferred embodiment, when the immunoglobulin is IgD, the value of the critical value of the IgD protein expression level for determining the high risk or low risk of colorectal cancer by the detection reagent is in the range of 1906.41pg/ml to 6168.47 pg/ml.

In a preferred embodiment, the critical value of the expression level of the IgD protein is 5842.19 pg/ml; when the expression quantity of the IgD protein is less than or equal to 5842.19pg/ml, the colorectal cancer risk is low; the detection result of the detection reagent is as follows: if the expression amount of the IgD protein is greater than 5842.19pg/ml, the colorectal cancer is at high risk.

In a preferred embodiment, when the immunoglobulin is IgE, the detection reagent determines that the critical value of the expression level of the IgE protein with high risk or low risk of colorectal cancer is 978.68pg/ml to 1305.34 pg/ml.

In a preferred embodiment, the critical value of the IgE protein expression amount is 991.94 pg/ml; when the expression quantity of the IgE protein is less than or equal to 991.94pg/ml, the colorectal cancer is at high risk; the detection result of the detection reagent is as follows: the expression level of IgE protein is greater than 991.94pg/ml, and the colorectal cancer risk is low.

In a preferred embodiment, when there are at least two immunoglobulins, the threshold value of the risk index X for determining a high risk or a low risk of colorectal cancer is in the range of 0.5000 to 0.7416.

As a preferred embodiment, when the immunoglobulins are IgD and IgE, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.00158C)_IgE+0.000131C_IgD+1.474))) of said C_IgEIs the concentration of IgE, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5920-0.6418; in a preferred embodiment, the critical value of the risk index X is 0.6039.

As a preferred embodiment, when the immunoglobulins are IgD, IgE and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001602C)_IgE+0.000119C_IgG3+0.0001C_IgD+1.091))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDThe concentration is the concentration of IgD, the concentration is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.5000-0.6762; in a preferred embodiment, the critical value of the risk index X is 0.5265.

As a preferred embodiment, when the immunoglobulins are IgD, IgE, IgG1 and IgG3, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001904C)_IgE+0.000273C_IgG3+0.000107C_IgD-0.000008C_IgG1+2.047))) of said C_IgEIs the concentration of IgE, C_IgG3Being IgG3Concentration of the C_IgDIs the concentration of IgD, said C_IgG1The concentration of IgG1 is the concentration of a test sample diluted by 10000 times, and the critical value range of the risk index X is 0.6231-0.6659; in a preferred embodiment, the critical value of the risk index X is 0.6280.

As a preferred embodiment, when the immunoglobulins are IgD, IgE, IgG3 and IgA1, the risk index X is calculated by: x is 1/(1+ e ^ (- (-0.001587C)_IgE+0.000212C_IgG3+0.000110C_IgD-0.000019C_IgA1+2.769))) of said C_IgEIs the concentration of IgE, C_IgG3At a concentration of IgG3, C_IgDIs the concentration of IgD, said C_IgA1The concentration of IgA1 is the concentration of a detection sample diluted by 10000 times, and the critical value range of the risk index X is 0.5933-0.7416; in a preferred embodiment, the critical value of the risk index X is 0.7374.

In a preferred embodiment, the detection result of the detection reagent is: when the risk index X is less than or equal to a critical value, the colorectal cancer is at low risk; when the risk index X > cutoff value, there is a high risk of colorectal cancer.

The invention has the beneficial effects that:

(1) the present inventors have found that among the numerous immunoglobulins, different immunoglobulins are differentially expressed and can serve as colorectal biomarkers.

(2) The detection object of the detection reagent is blood, and dynamic and large-scale screening can be realized.

(3) Compared with other proteins in plasma, the immunoglobulin has longer half-life and stable property, and is more suitable to be used as a marker for disease diagnosis.

Drawings

Figure 1 is a differential immunoglobulin volcano plot.

FIG. 2 is a comparison of the plasma concentrations of different immunoglobulins in 51 healthy persons and 99 colorectal cancer patients.

FIG. 3 ROC curve analysis of different immunoglobulin indices to differentiate colorectal cancers.

FIG. 4 is a graph of the gradient dilution of an immunoglobulin standard.

Detailed Description

The technical solutions of the present invention are further illustrated by the following specific examples, which do not represent limitations to the scope of the present invention. Insubstantial modifications and adaptations of the present invention by others of the concepts fall within the scope of the invention.