CN117031004B - Application of reagent for detecting omelanin-1 in preparation of product for predicting or evaluating quality of life of IBS patient - Google Patents
Application of reagent for detecting omelanin-1 in preparation of product for predicting or evaluating quality of life of IBS patient Download PDFInfo
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Abstract
The invention discloses an application of a reagent for detecting omelanin-1 in preparing a product for predicting or evaluating the life quality of IBS patients, and relates to the field of gastrointestinal diseases. The invention discovers for the first time that the quality of life evaluation score of IBS patients is inversely related to the concentration of omelanin-1 in serum or plasma of IBS patients, and the difference is significant. The lower the concentration of omelanin-1 in serum or plasma, the higher the quality of life evaluation score of IBS patients, the poorer the self-perceived quality of life. Therefore, in the dynamic management of IBS patients, the change of serum omelanin-1 concentration has better clinical application prospect for predicting or evaluating the quality of life of IBS. In addition, the method for directly detecting the concentration of the omelanin-1 in serum or plasma has the technical advantages of objective result, convenient sample acquisition and minimally invasive. The invention provides a new means for managing the dynamic change of the quality of life of IBS patients clinically, and has better clinical application prospect.
Description
Technical Field
The invention relates to the field of gastrointestinal diseases, in particular to application of a reagent for detecting omelanin-1 in preparation of a product for predicting or evaluating quality of life of IBS patients.
Background
Irritable bowel syndrome (irritable bowel syndrome, IBS) is one of the most common gastrointestinal diseases. IBS belongs to a class of functional gastrointestinal disorders, and current diagnosis of IBS is mainly based on clinical symptoms of patients, excluding other organic diseases, but lacks specific biomarkers or clinical indicators as the gold standard for diagnosis. Treatment of IBS has heretofore remained a clinical challenge. IBS patients often complain of gastrointestinal symptoms of various unknown causes that have varying degrees of impact on the patient's daily life. Almost half of patients have a need to seek medical assistance, even multiple times, resulting in significant direct and indirect social medical costs.
The general impact of multiple factors is particularly important from the perspective of the patient, as it lacks specific diagnostic indicators for IBS, and various social culture, environmental and behavioral factors are important for the development and progression of IBS. Studies have shown that IBS patients consume high medical costs, frequently seek medical attention and leave, and, as with organic disease patients, may exhibit impaired health.
Furthermore, an increasing number of people believe that health-related quality of life (QOL) assessment should be an important component of clinical research and therapeutic trials.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide application of a reagent for detecting omelanin-1 in preparation of products for predicting or evaluating quality of life of IBS patients, so as to realize prediction of quality of life of IBS patients, in particular prediction or evaluation of health-related quality of life.
The invention is realized in the following way:
In a first aspect, the invention provides an application of a reagent for detecting omelanin-1 in preparing a kit for predicting or evaluating the life quality of IBS patients, wherein the kit takes the omelanin-1 as a detection marker.
The inventors have found for the first time that the (IBS-QOL) score of quality of life assessment for IBS patients is inversely related to the concentration of omelanin-1 (Omentin-1) in serum or plasma of IBS patients, and that the difference is significant. The lower the Omentin-1 concentration in serum or plasma, the higher the quality of life evaluation score of the IBS patient, and the worse the self-perceived quality of life. Therefore, in the dynamic management of IBS patients, the change of serum Omentin-1 concentration has better clinical application prospect for predicting or evaluating the quality of life of IBS.
In addition, the result of Omentin-1 concentration detection of serum or plasma is objective, and the serum or plasma sample is convenient to obtain and can be obtained through minimally invasive. The invention provides a new means for managing the dynamic change of the quality of life of IBS patients clinically, and has better clinical application prospect.
The kit can be a commercially available omelanin-1 detection kit, or a self-made omelanin-1 detection kit.
In a preferred embodiment of the invention, the kit is for: the increase in serum omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients. In an alternative embodiment, a significant improvement in quality of life in IBS patients is predicted or assessed by an increase in the concentration of omelanin-1 in serum of IBS patients of at least 30% as compared to the detection results several days ago, several months ago (e.g., 1 month, 2 months, 3 months, or 6 months), 1 year ago, or years ago. Increases in concentration of omestatin-1 by at least 30% over previous test concentrations include, but are not limited to: 30% -35% raised, 30% -40% raised, or 40% -60% raised, or 60% -100% raised.
The prediction refers to prediction or prediction of the quality of life of an IBS patient, and may predict the influence of various social culture, environmental and behavioral factors on the occurrence and/or development of IBS.
Evaluation refers to: assessment of the quality of life of IBS patients, the impact of various social culture, environmental and behavioral factors, etc., on the occurrence and/or development of IBS can be assessed, including but not limited to those that have already been produced, those that are about to or are expected to be produced. In one embodiment, the risk of impact of various social culture, environmental and behavioral factors on the development of IBS may be assessed, thereby providing a reference for subsequent treatment or prognosis.
The serum is selected from, for example, the serum of peripheral blood.
In a preferred embodiment of the invention, the kit is for: the increase in plasma omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
In a preferred embodiment of the invention, the agent for detecting omelanin-1 is selected from antibodies against omelanin-1. Such as monoclonal antibodies against omelanin-1.
In an alternative embodiment, the kit further comprises: ELISA plate, washing solution (or rinsing solution), enzyme-labeled antibody and color development solution. The enzyme-labeled antibody includes, but is not limited to, an enzyme labeled on an antibody that catalyzes the development of a substrate.
Enzymes that catalyze the development of a substrate include, but are not limited to, horseradish peroxidase, alkaline phosphatase, beta-galactosidase, glucose oxidase, carbonic anhydrase, acetylcholinesterase, and glucose-6-phosphate deoxygenase.
In a second aspect, the invention also provides application of the reagent for detecting the omelanin-1 in preparing a chip for predicting or evaluating the life quality of IBS patients, wherein the chip takes the omelanin-1 as a detection marker.
In a preferred embodiment of the invention, the chip is for: the increase in serum omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
In a preferred embodiment of the invention, the chip is for: the increase in plasma omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
In a preferred embodiment of the invention, the agent for detecting omelanin-1 is selected from antibodies against omelanin-1.
In a third aspect, the invention also provides application of the reagent for detecting the omelanin-1 in preparing a test strip for predicting or evaluating the life quality of IBS patients, wherein the test strip takes the omelanin-1 as a detection marker.
In an alternative embodiment, the antibodies on the test strip are labeled with a nanoparticle-based label. Nanoparticle-based labels include, but are not limited to, nanoparticles and colloids.
Nanoparticles include, but are not limited to: organic nanoparticles, magnetic nanoparticles, quantum dot nanoparticles, and rare earth complex nanoparticles.
In alternative embodiments, colloids include, but are not limited to, colloidal metals, disperse dyes, dye-labeled microspheres, and latex.
In alternative embodiments, the colloidal metals include, but are not limited to, colloidal gold, colloidal silver, and colloidal selenium.
In a preferred embodiment of the invention, the test strip is used for: the quality of life of the IBS patient is predicted or assessed to be significantly improved by increasing the concentration of omelanin-1 in serum or plasma of the IBS patient by at least 30% as compared to the previous test results.
In a fourth aspect, the invention also provides the use of a reagent for detecting omelanin-1 in the preparation of a system for predicting or assessing the quality of life of a patient with IBS, wherein the system uses omelanin-1 as a detection marker.
In a preferred embodiment of the invention, the system is used for: the quality of life of the IBS patient is predicted or assessed to be significantly improved by increasing the concentration of omelanin-1 in serum or plasma of the IBS patient by at least 30% as compared to the previous test results.
In a preferred embodiment of the application of the invention, the system comprises a sample loading module, a detection module and an analysis module. The system can realize the content detection of the omelanin-1 in the sample in a semi-automatic or automatic way.
The sample loading module is used for loading samples to the detection module; the detection module is used for detecting the content of the omelanin-1, including but not limited to: antigen coated plate, sealing, primary antibody incubation, washing, enzyme-labeled antibody incubation, washing and color development; analysis modules include, but are not limited to: and traversing the detection results before traversing, and comparing and analyzing to obtain the quality of life result.
Such systems include, but are not limited to, dynamic monitoring systems, traceability systems, and the like.
The invention has the following beneficial effects:
The invention discovers for the first time that the (IBS-QOL) score of the quality of life evaluation of IBS patients is inversely related to the concentration of omelanin-1 (Omentin-1) in serum or plasma of IBS patients, and the difference is significant. The lower the Omentin-1 concentration in serum or plasma, the higher the quality of life evaluation score of the IBS patient, and the worse the self-perceived quality of life. Therefore, in the dynamic management of IBS patients, the change of serum Omentin-1 concentration has better clinical application prospect for predicting or evaluating the quality of life of IBS.
In addition, the result of Omentin-1 concentration detection of serum or plasma is objective, and the serum or plasma sample is convenient to obtain and can be obtained through minimally invasive. The invention provides a new means for managing the dynamic change of the quality of life of IBS patients clinically, and has better clinical application prospect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph of statistical results of Omentin-1 concentrations in serum of IBS patients and healthy Controls (Controls) (IBS patients, 150; healthy Controls, 150. P <0.001, statistical test MANN WHITNEY TEST);
FIG. 2 is a graph of statistical results of correlation analysis between serum Omentin-1 and IBS-QOL score of IBS patients ((A) study design; B) analysis of correlation between serum Omentin-1 and IBS-QOL score of IBS patients, 150 of which p <0.001, statistical test using Pearson's correlation analysis);
FIG. 3 is a graph of quality of life improvement in IBS patients versus dynamic serum Omentin-1 concentration monitoring (105 patients included; p <0.001, and statistical test using Chi-square test).
Detailed Description
Reference now will be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the invention. Indeed, it will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment can be used on another embodiment to yield still a further embodiment.
Unless otherwise indicated, practice of the present invention will employ conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry and immunology, which are within the ability of a person skilled in the art. This technique is well explained in the literature, as is the case for molecular cloning: laboratory Manual (Molecular Cloning: ALaboratory Manual), second edition (Sambrook et al, 1989); oligonucleotide Synthesis (Oligonucleotide Synthesis) (M.J.Gait, eds., 1984); animal cell Culture (ANIMAL CELL Culture) (r.i. freshney, 1987); methods of enzymology (Methods in Enzymology) (academic Press Co., ltd. (ACADEMIC PRESS, inc.)), experimental immunology handbook (Handbook of Experimental Immunology) (D.M.Weir and C.C. Blackwell, inc.), gene transfer Vectors for mammalian cells (GENE TRANSFER vector for MAMMALIAN CELLS) (J.M.Miller and M.P.Calos, inc., 1987), methods of contemporary molecular biology (Current Protocols in Molecular Biology) (F.M.Ausubel et al, 1987), polymerase chain reaction (PCR: the Polymerase Chain Reaction) (Mullis et al, 1994), and methods of contemporary immunology (Current Protocols in Immunology) (J.E.Coligan et al, 1991), each of which are expressly incorporated herein by reference.
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
The features and capabilities of the present invention are described in further detail below in connection with the examples.
Example 1
This example compares Omentin-1 concentrations in serum from IBS patients and healthy Controls (Controls) and performs quality of life analysis.
1) Personnel entering group
This example recruits 150 patients diagnosed with diarrhea type IBS according to roman III diagnostic criteria at the people hospital in the province of Sichuan. In addition, 150 healthy volunteers of the control group matched in gender, age, and marital status were enrolled.
The criteria for inclusion in the group include evidence of infection, celiac disease, other digestive tract diseases such as inflammatory bowel disease, abdominal surgery, chronic diseases such as diabetes, smoking, excessive alcohol consumption, use of steroid and non-steroid anti-inflammatory drugs. In all control subjects, irritable bowel syndrome and other functional bowel disorders were excluded by medical interrogation. In addition, none of the participants took medications that were known to affect the gastrointestinal tract or immune system.
2) Quality of life assessment for IBS patients
The quality of life of IBS patients was evaluated using a system widely used in IBS studies, called IBS-QOL questionnaire (see publication Drossman DA,Patrick DL,Whitehead WE,et al.Further validation of the IBS-QOL:a disease-specific quality-of-life questionnaire.Am J Gastroenterol2000;95:999-1007). The questionnaire includes 34 questions, each of 0-5 points, each of which the participants answer to all 34 items, and aggregate the scores, which then translate to a metering range of 100 points.
3) Serum Omentin-1 concentration detection
The Omentin-1 detection kit was purchased from BioLegend (San Diego, CA, USA) using an enzyme-linked immunosorbent assay (ELISA). The detailed operation steps are as follows:
(1) Antigen coating (i.e. serum coating): the antigen was diluted to 10mg/ml with carbonate coating solution, 100ml was added to each microplate well, and the mixture was placed in a refrigerator at 4℃overnight. The antigen liquid was aspirated, and each well was washed with 100ml PBST, once for 5 min.
(2) Closing: 100ml of blocking solution (5% BSA) was added to each of the microplate wells, incubated at room temperature for 60min, the blocking solution was removed by pipetting, and washed with PBST in a volume of 100ml for 5 min.
(3) Incubation resistance: the washing solution was removed, and a primary antibody (anti-omelanin-1 antibody) diluted 500 to 1000 times with 5% BSA was added to each of the wells, and the wells were incubated at room temperature for 1 hour in a volume of 100 ml.
(4) Washing: the liquid in the wells of the enzyme-labeled wells was aspirated, and each well was washed three times with PBST in a volume of 100ml per well for 5min.
(5) Adding enzyme-labeled antibody: the wash was blotted clean and 100ml of secondary antibody diluted 1:2000 with 5% BSA was added to each well and incubated at room temperature for 1h.
(6) Washing: the liquid in the wells was removed and washed three times with PBST for 5min each with 100ml per well volume, and then 100ml PBS was added to each well for a further 5min.
(7) Color development: 100mlTMB substrate solutions are added into each hole, and the reaction is carried out for 40min at normal temperature in the dark state in the liquid adding process.
(8) Terminating the reaction: the reaction was stopped by adding 100ml of 2M sulfuric acid to each microplate.
(9) Reading a plate: the microplate was placed on the microplate reader for reading.
By analyzing the peripheral venous serum Omentin-1 concentrations of 150 IBS patients and 150 healthy control groups, omentin-1 concentration in the serum of IBS patients was found to be significantly lower than that of healthy controls (as shown in fig. 1).
Quality of life analysis was performed on patients enrolled in IBS by the IBS-QOL system (as shown in FIG. 2A), and the IBS-QOL score was found to be inversely related to the Omentin-1 concentration in the serum of IBS patients, with the difference being significant (as shown in FIG. 2B), suggesting that the lower the Omentin-1 concentration in the serum of IBS patients, the higher the IBS-QOL score, the poorer the self-perceived quality of life.
Example 2
For 105 of the above-incorporated patients, a dynamic follow-up study was performed for 1 year, with a specific protocol grouping scheme as seen in fig. 3.
According to the change of Omentin-1 concentration in serum of IBS patients after 1 year of follow-up, ① groups were elevated by 30% or more and ② groups were changed by less than 30%. A decrease in IBS-QOL score of 10 or greater is defined as a significant improvement in quality of life for IBS patients.
The number of patients with significantly improved quality of life for IBS patients in both groups of people was analyzed separately. Through statistical analysis, the number of patients (① groups) with the concentration of Omentin-1 in serum increased by more than or equal to 30 percent, the life quality of which is obviously improved (42/58 people) is obviously higher than the number of people (11/47 people) with the concentration of Omentin-1 in serum increased by less than 30 percent (② groups), which indicates that in the dynamic management of IBS patients, the change of the concentration of the serum Omentin-1 has better clinical application prospect for detecting the improvement of the life quality of IBS.
In conclusion, the invention discovers a newly discovered fatty substance-Omentin-1 for the first time, and has good efficacy in predicting the life quality of IBS patients. The concentration of Omentin-1 detected in serum or plasma has the technical advantages of objective result, convenient sample acquisition and minimally invasive. The invention provides a new means for managing the dynamic change of the quality of life of IBS patients clinically, and has better clinical application prospect.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (13)
1. The application of a reagent for detecting omelanin-1 in preparing a kit for predicting or evaluating the life quality of IBS patients is characterized in that the kit takes the omelanin-1 as a detection marker.
2. The use according to claim 1, wherein the kit is for: the increase in serum omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
3. The use according to claim 1, wherein the kit is for: the increase in plasma omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
4. The use according to any one of claims 1 to 3, wherein the reagent for detecting omelanin-1 is selected from antibodies against omelanin-1.
5. The application of a reagent for detecting omelanin-1 in preparing a chip for predicting or evaluating the life quality of IBS patients is characterized in that the chip takes the omelanin-1 as a detection marker.
6. The use of claim 5, wherein the chip is configured to: the increase in serum omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
7. The use of claim 5, wherein the chip is configured to: the increase in plasma omelanin-1 concentration of IBS patients by at least 30% compared to the previous test results predicts or evaluates a significant improvement in quality of life for IBS patients.
8. The use according to any one of claims 5 to 7, wherein the reagent for detecting omelanin-1 is selected from antibodies against omelanin-1.
9. The application of a reagent for detecting omelanin-1 in preparing a test strip for predicting or evaluating the life quality of IBS patients is characterized in that the test strip takes the omelanin-1 as a detection marker.
10. The use of claim 9, wherein the test strip is for: the quality of life of the IBS patient is predicted or assessed to be significantly improved by increasing the concentration of omelanin-1 in serum or plasma of the IBS patient by at least 30% as compared to the previous test results.
11. Use of a reagent for detecting omelanin-1 in the preparation of a system for predicting or assessing the quality of life of a patient with IBS, wherein the system uses omelanin-1 as a detection marker.
12. The use according to claim 11, wherein the system is for: the quality of life of the IBS patient is predicted or assessed to be significantly improved by increasing the concentration of omelanin-1 in serum or plasma of the IBS patient by at least 30% as compared to the previous test results.
13. The use of claim 12, wherein the system comprises a sample module, a detection module, and an analysis module.
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| CN102639710A (en) * | 2009-07-20 | 2012-08-15 | 基因泰克公司 | Gene expression markers for crohn's disease |
| CN115248312A (en) * | 2021-04-27 | 2022-10-28 | 昆山美卡生物医药科技有限公司 | Method for detecting serum DNA |
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| US10413588B2 (en) * | 2015-11-05 | 2019-09-17 | Wisconsin Alumni Research Foundation | Probiotic function of human intelectin |
| CN112485448B (en) * | 2020-11-20 | 2021-08-24 | 浙江大学 | Application of NMI protein as biological target in the preparation of adult community-acquired pneumonia severity and prognosis assessment kit |
| US20220233119A1 (en) * | 2021-01-22 | 2022-07-28 | Ethicon Llc | Method of adjusting a surgical parameter based on biomarker measurements |
| CN116036072A (en) * | 2023-02-10 | 2023-05-02 | 四川省医学科学院·四川省人民医院 | Application of isoginkgetin in preparation of medicines for preventing and/or treating ulcerative colitis |
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| CN102639710A (en) * | 2009-07-20 | 2012-08-15 | 基因泰克公司 | Gene expression markers for crohn's disease |
| CN115248312A (en) * | 2021-04-27 | 2022-10-28 | 昆山美卡生物医药科技有限公司 | Method for detecting serum DNA |
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