CN111007258A - Reagent for early diagnosis of Kawasaki disease and application thereof - Google Patents
Reagent for early diagnosis of Kawasaki disease and application thereof Download PDFInfo
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- CN111007258A CN111007258A CN201911326000.XA CN201911326000A CN111007258A CN 111007258 A CN111007258 A CN 111007258A CN 201911326000 A CN201911326000 A CN 201911326000A CN 111007258 A CN111007258 A CN 111007258A
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Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
Abstract
The invention provides a reagent for early diagnosis of children Kawasaki disease and application thereof. The invention also provides application of the reagent in preparation of a kit for early diagnosis of Kawasaki disease. By detecting the content of thrombospondin in the plasma of the suspected child to assist in diagnosing the Kawasaki disease, the suspected child can be diagnosed and treated early, so that the occurrence of pathological changes such as cardiovascular diseases and the like can be prevented.
Description
Technical Field
The invention belongs to the field of medicine, and particularly relates to a reagent for early diagnosis of Kawasaki disease and application thereof, in particular to a reagent for detecting thrombospondin in plasma, and application of the reagent in preparation of a kit for early diagnosis of Kawasaki disease.
Technical Field
Kawasaki Disease (KD) occurs well in children under the age of 5 years, and although the cause of the disease is not clear, the occurrence of the disease is generally considered to be related to abnormal immune response. The pathological change is systemic vasculitis, which can affect arteries, veins and capillaries, especially coronary artery damage is most obvious, and severe cases can cause coronary aneurysm, myocardial infarction and sudden cardiac death. Currently, along with the improvement of the curative effect and prognosis of infectious diseases, kawasaki disease has replaced rheumatic fever and becomes the most common cause of acquired heart disease of children. Early diagnosis and timely treatment of the disease are attracting more and more attention.
TSP of higher animals is classified into two groups, group A includes TSP-1 and TSP-2, group B includes TSP-3, TSP-4 and COMP/TSP-5, wherein TSP-1 is an extracellular matrix glycoprotein secreted by platelet α granules, fibroblasts, vascular endothelial cells, etc., TSP-1 and TSP-2 are involved in important physiological processes of neovascularization, tissue repair, etc., and are closely related to the occurrence and development of various diseases.
At present, KD is diagnosed mainly based on clinical manifestations, but not specifically. Clinically there is also incomplete KD, and diseases that require differential diagnosis include measles, other viral infections (e.g., adenovirus, enterovirus), staphylococcal and streptococcal infections, scarlet fever, and the like. Although KD is a self-limiting disease, most patients have good prognosis, but few patients still have complications, particularly cardiovascular complications, and serious patients can have myocardial insufficiency, cardiovascular failure and the like, so early diagnosis and timely treatment of kawasaki disease have important significance for prognosis.
Disclosure of Invention
In order to overcome the defects of the prior art, clinical observation and verification show that thrombospondin 1(thrombospondins-1, TSP-1) and thrombospondin 2(thrombospondins-2, TSP-2) are related to acute phase inflammatory reaction of Kawasaki disease and can be used for early diagnosis of the Kawasaki disease, so that the invention provides the following steps:
a detection reagent for detecting thrombospondin 1(thrombospondins-1, TSP-1), and/or thrombospondin 2(thrombospondins-2, TSP-2).
Preferably, the detection reagent is used for detecting TSP-1.
More preferably, the detection reagent is used for detecting TSP-1 and TSP-2.
More preferably, the detection reagent is an antibody against TSP-1 and/or TSP-2 described above.
The invention also provides application of the detection reagent in preparation of a kit for early diagnosis of Kawasaki disease.
The invention also provides application of the detection reagent in preparation of a reagent kit for early diagnosis of Kawasaki disease in combination with other reagents, wherein the other reagents are reagents for detecting indexes of Kawasaki Disease (KD) in a laboratory.
Preferably, the KD laboratory measures include, but are not limited to, total white blood cells, percent neutrophils, serum sodium (Na), serum albumin, serum total bilirubin, C-reactive protein, Erythrocyte Sedimentation Rate (ESR), platelets, transaminases, aseptic pus, cerebrospinal fluid mononuclear cells, and the like.
More preferably, the KD laboratory measures include C-reactive protein, neutrophil percentage, serum sodium (Na), serum albumin, serum total bilirubin, and the like.
The invention also provides a kit, which comprises the detection reagent and is used for early diagnosis of Kawasaki disease.
Preferably, the kit further comprises a reagent for detecting Kawasaki Disease (KD) laboratory indexes.
Preferably, the KD laboratory measures include, but are not limited to, total white blood cells, percent neutrophils, serum sodium (Na), serum albumin, serum total bilirubin, C-reactive protein, Erythrocyte Sedimentation Rate (ESR), platelets, transaminases, aseptic pus, cerebrospinal fluid mononuclear cells, and the like.
More preferably, the KD laboratory measures include C-reactive protein, neutrophil percentage, serum sodium (Na), serum albumin, serum total bilirubin, and the like.
The invention also provides a method for detecting TSP-1 and/or TSP-2, which comprises the step of detecting a sample by using the detection reagent.
Preferably, the sample is derived from blood. Preferably, the sample is derived from plasma.
Preferably, the method comprises the steps of:
(1) processing a sample; preferably, the sample is derived from plasma.
(2) Enzyme-linked immunoassay of TSP-1 and/or TSP-2;
(3) and (6) analyzing results.
The invention has the advantages that: the invention firstly determines the index for early diagnosis of Kawasaki disease through clinical observation and biochemical detection, and has important significance for early diagnosis, early treatment, prevention of the occurrence of cardiovascular and other pathological changes and the like. And the kit has the characteristics of simple and convenient operation, low cost and the like.
Drawings
Embodiments of the invention are described in detail below with reference to the attached drawing figures, wherein:
FIG. 1: changes of plasma TSP-1 and TSP-2 levels of children suffering from Kawasaki disease in acute stage and control group
Detailed Description
The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Example 1: detection of TSP-1 and/or TSP-2 in Kawasaki disease plasma
Data and method
1. General data:
the study was divided into kawasaki disease group and control group. The Kawasaki disease group comprises 51 patients who are firstly diagnosed with Kawasaki disease according to diagnosis standards in hospital of children affiliated to the first institute of pediatrics in 2009-2012 3, wherein 33 men and 18 women have a male-female ratio of 1.8: 1, age of 0.25-8.9 years and average age of 2.2 years. The group is divided into 2 subgroups according to heart affected conditions, wherein the heart affected group is examined by an echocardiogram, an electrocardiogram, a myocardial zymogram and the like, and one abnormal person is detected; the group without any abnormality was not heart affected. The control group consisted of 21 individuals, 16 individuals male and 5 individuals female, with a male-female ratio of 3.2: 1, age 0.42-10.8 years, and average age 4.7 years. The group is divided into a heating group and a non-heating group. The fever group includes 12 patients with contemporary age-matched non-systemic inflammatory response fever (upper respiratory tract infection, pneumonia, etc.), and the non-fever group includes 9 healthy children. After inquiring about clinical medical history, physical examination and laboratory examination, all the infants in the control group except patients with Kawasaki disease, cardiovascular diseases and other related medical history.
2. Sample treatment:
all the study subjects were treated after admission and before starting treatment, 3ml of fasting venous blood was collected, injected into a closed blood collection tube containing ethylenediaminetetraacetic acid (EDTA) anticoagulant, centrifuged at 3350 Xg for 15min, serum was separated, and stored in a freezer at-80 ℃. And simultaneously, measuring indexes such as peripheral blood myocardial zymogram and the like, and performing electrocardiogram and echocardiography examination to evaluate the functional conditions of coronary arteries and heart. The coronary artery disease standard adopts the formulated standard of 7 th edition of Zhufu Tang Utility Ergonomics.
Determination of TSP content:
the determination of the content of TSP-1 and TSP-2 is carried out by adopting a human thrombospondin enzyme-linked immunoassay kit (Beijing Cork-Advance science and technology center, Code No.1027969, No.1027968) according to the steps of a kit specification.
4. Statistical treatment:
SPSS16.0 statistical software is applied, TSP-1 with various metering parameters conforming to normal distribution is represented by mean plus or minus standard deviation (x plus or minus s), and mean comparison among groups adopts independent sample t test; the TSP-2 with the off-normal distribution is expressed by median (P25, P75), and the difference of P < 0.05 is statistically significant by adopting the rank sum test in comparison among groups.
Second, result in
The changes of TSP-1 and TSP-2 in the acute stage of children with Kawasaki disease are that the plasma TSP-1 level of the children with Kawasaki disease in the acute stage is obviously increased compared with that of a control fever group and a control non-fever group, the difference has statistical significance (P is less than 0.01), the plasma TSP2 level of the children with Kawasaki disease is obviously increased compared with that of the control fever group (P is less than 0.05), the control fever group has a rising trend (P is more than 0.05), and the change difference of TSP-1 and TSP-2 does not have statistical significance (P is more than 0.05) compared with that of the control fever group. In particular, see fig. 1, wherein compared to kawasaki disease group,aP＝0.000，bP＝0.020，cp is 0.218, compared to the control fever group,dP＝0.228，eP＝0.111。
the application finds that the TSP-1 and TSP-2 levels are obviously increased in the acute stage of Kawasaki patients compared with other non-systemic inflammatory response fever or non-pyrogenic diseases. The change of the plasma TSP level is suggested to possibly participate in early systemic inflammatory response of the Kawasaki disease, and the TSP can be used as a prediction factor of systemic immune inflammatory response diseases such as the Kawasaki disease and the like, and has important clinical significance for early differential diagnosis.
Due to disturbance of immune system and vascular matrix metabolism in Kawasaki patients, the increase of TSP-1 and TSP-2 levels may play a protective role in acute stage of Kawasaki disease to inhibit immune response, MMP activation and VEGF increase. In the study, the stability of TSP-1 is higher than that of TSP-2, the individual difference of TSP-2 is large, the TSP-1 protection effect is stronger in Kawasaki disease, and the TSP-1 can be better used as a predictor of systemic inflammatory response diseases.
The clinical significance of the research is that TSP can be used as an index of systemic immune inflammatory response under the condition that clinical manifestations of fever, rash and Kawasaki disease diagnosis standards are insufficient, and early differential diagnosis is facilitated.
Meanwhile, TSP-1 and TSP-2 of the present invention may be used for early diagnosis in conjunction with other laboratory tests including, but not limited to, total white blood cells, percentage of neutrophils, serum sodium (Na), serum albumin, serum total bilirubin, C-reactive protein, Erythrocyte Sedimentation Rate (ESR), platelets, transaminase, aseptic purulent urine, cerebrospinal fluid mononuclear cells, wherein kawasaki patients exhibit increased total white blood cell count, increased percentage of neutrophils, decreased serum sodium, hypoalbuminemia, increased serum total bilirubin, increased C-reactive protein (CRP), increased Erythrocyte Sedimentation Rate (ESR), increased platelets, increased transaminase, aseptic purulent urine, cerebrospinal fluid mononuclear cells, and the like.
Preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, and these simple modifications all belong to the protection scope of the invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
Claims (10)
1. A detection reagent for detecting thrombospondin 1(thrombospondins-1, TSP-1), and/or thrombospondin 2(thrombospondins-2, TSP-2).
2. The detection reagent according to claim 1, wherein the detection reagent is used for detecting TSP-1.
3. The detection reagent of claim 1, wherein the detection reagents TSP-1 and TSP-2.
4. The detection reagent according to any one of claims 1 to 3, wherein the detection reagent is an antibody against the above TSP-1 and/or TSP-2.
5. Use of the detection reagent according to any one of claims 1-4 in the preparation of a kit for early diagnosis of Kawasaki disease.
6. Use of the detection reagent according to any one of claims 1-4 in combination with other reagents for preparing a kit for early diagnosis of Kawasaki disease, wherein the other reagents are reagents for detecting laboratory indicators of Kawasaki disease.
7. The use of claim 6, wherein the laboratory metrics include, but are not limited to, total white blood cells, percent neutrophils, serum sodium (Na), serum albumin, serum total bilirubin, C-reactive protein, Erythrocyte Sedimentation Rate (ESR), platelets, transaminase, aseptic pus, cerebrospinal fluid mononuclear cells.
8. The use as claimed in any one of claims 5 to 7, wherein the TSP-1 and TSP-2 content is increased for early diagnosis of Kawasaki disease.
9. A kit comprising the detection reagent according to any one of claims 1 to 4.
10. The kit of claim 9, further comprising reagents for detecting a laboratory indicator of Kawasaki Disease (KD).
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| CN201911326000.XA CN111007258A (en) | 2019-12-20 | 2019-12-20 | Reagent for early diagnosis of Kawasaki disease and application thereof |
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| CN201911326000.XA CN111007258A (en) | 2019-12-20 | 2019-12-20 | Reagent for early diagnosis of Kawasaki disease and application thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112630431A (en) * | 2020-10-26 | 2021-04-09 | 上海柏运医疗器材有限公司 | Application of human thrombin sensitive protein-1 in preparation of kit for predicting chemotherapy effect of intrahepatic bile duct cancer |
| CN116773825A (en) * | 2022-08-19 | 2023-09-19 | 天津云检医学检验所有限公司 | Blood biomarkers and methods for diagnosing acute kawasaki disease |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112630431A (en) * | 2020-10-26 | 2021-04-09 | 上海柏运医疗器材有限公司 | Application of human thrombin sensitive protein-1 in preparation of kit for predicting chemotherapy effect of intrahepatic bile duct cancer |
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