[go: up one dir, main page]

CN111443201A - Aspirin drug resistance detection kit - Google Patents

Aspirin drug resistance detection kit Download PDF

Info

Publication number
CN111443201A
CN111443201A CN202010114209.6A CN202010114209A CN111443201A CN 111443201 A CN111443201 A CN 111443201A CN 202010114209 A CN202010114209 A CN 202010114209A CN 111443201 A CN111443201 A CN 111443201A
Authority
CN
China
Prior art keywords
thromboxane
reagent
dehydro
aspirin
creatinine
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202010114209.6A
Other languages
Chinese (zh)
Inventor
张伯平
蔡泽浪
黄夏雨
邹佳信
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen Adibio Technology Co ltd
Original Assignee
Shenzhen Adibio Technology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen Adibio Technology Co ltd filed Critical Shenzhen Adibio Technology Co ltd
Priority to CN202010114209.6A priority Critical patent/CN111443201A/en
Publication of CN111443201A publication Critical patent/CN111443201A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90Enzymes; Proenzymes
    • G01N2333/902Oxidoreductases (1.)
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00Detection or diagnosis of diseases
    • G01N2800/28Neurological disorders
    • G01N2800/2871Cerebrovascular disorders, e.g. stroke, cerebral infarct, cerebral haemorrhage, transient ischemic event

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention provides an aspirin drug resistance detection kit, which is used for detecting 11-dehydro-thromboxane B in human urine2The ratio of the aspirin and the creatinine is used for judging the aspirin resistance degree of a patient, and the method is characterized in that a homogeneous enzyme immunoassay method is adopted for measuring 11-dehydro-thromboxane B in human urine2The content of the (D) has the characteristics of no wound, high automation degree, rapid detection, low cost and the like; the kit simultaneously comprises a urinary creatinine detection reagent, the two detections can be simultaneously carried out on a full-automatic biochemical analyzer, and the detection result can be automatically calculated.

Description

Aspirin drug resistance detection kit
Technical Field
The invention belongs to the field of in-vitro diagnosis, and particularly relates to an aspirin resistance detection kit which is used for determining aspirin resistance of a human body.
Background
Cardiovascular and cerebrovascular diseases caused by thrombus become one of the main diseases endangering the health of people, and have the characteristics of high morbidity, high disability rate, high death rate and high recurrence rate. At present, evidence of evidence-based medicine at home and abroad proves that the continuous use of antithrombotic drugs and the effective control of risk factors are very critical to the improvement of the prognosis of patients. Aspirin is a salicylic acid non-steroidal anti-inflammatory drug, and can achieve an antithrombotic effect by intervening platelet aggregation. Recent guidelines set forth by the american heart association and the american stroke association emphasize that aspirin is an important first-line treatment and second-line prophylactic for patients with acute myocardial infarction. China 'diagnosis and treatment guidelines for acute myocardial infarction' also clearly points out that: as an antiplatelet therapy, all patients with acute myocardial infarction without contraindications should take water-soluble aspirin or chewable enteric aspirin 300mg immediately, and then maintain the aspirin for a long time at 75-100 mg/d; and after the patient is discharged from hospital, 75-100 mg/d of aspirin should be taken for a long time.
However, the therapeutic effect of aspirin is very limited for some patients. According to statistics, about 15-25% of patients with arterial thrombosis still have platelet activation and aggregation to form vascular embolism when taking aspirin for treatment, and the condition is called high aspirin platelet reaction and also called aspirin resistance. Therefore, there is a need for a reliable way for clinicians to analyze an individual's specific response to aspirin to assist the physician in determining treatment and adjusting the type and dosage of the individual accordingly. Studies have shown that 11-dehydro-thromboxane B is present in urine2(11-dehydro-thromboxane B2,11-dhTXB2) The ratio of the content to the creatinine content can accurately reflect the treatment effect of patients after taking aspirin, can predict the risk of cardiovascular and cerebrovascular accidents of the patients, and has extremely high clinical value.
11-dhTXB2Is one of metabolites of arachidonic acid in human urine, belongs to a small molecular compound, and has concentration related to water intake of patients, usually 300-5000 pg/ml. Due to 11-dhTXB in urine2The content is extremely low, and the urine components are complex and the number of interferents is large, so that the quantitative measurement of the urine is very difficult clinically. At present, the detection methods which can be practically applied are a chromatography method and an enzyme-linked immunosorbent assay, and the two detection methods have the disadvantages of complex operation process, long detection period, expensive equipment and manual operation, so that the application of the item is limited.
In summary, in the aspect of accurate aspirin administration, a detection method and a detection reagent which can be developed on a large scale and are low in cost are urgently needed clinically.
Disclosure of Invention
The invention provides an aspirin drug resistance detection kit, which is used for detecting 11-dehydro-thromboxane B in human urine2The ratio of the aspirin and the creatinine is used for judging the aspirin drug resistance degree of a patient, and the method is characterized in that a homogeneous enzyme immunoassay method is adopted for measuring 11-deaths in human urineHydro-thromboxane B2The content of the (D) has the characteristics of no wound, high automation degree, rapid detection, low cost and the like; the kit simultaneously comprises a urinary creatinine detection reagent, the two detections can be simultaneously carried out on a full-automatic biochemical analyzer, and the detection result can be automatically calculated.
The content of the invention comprises:
[1]an aspirin drug resistance detection kit, which is used for detecting 11-dehydro-thromboxane B in human urine2The ratio of the aspirin to the creatinine is used for judging the drug resistance of the patient to the aspirin; the kit consists of 11-dehydro-thromboxane B2The determination part, the creatinine determination part, the standard substance and the quality control substance are composed of 11-dehydro-thromboxane B in a kit2A homogeneous enzyme immunoassay method is used in the determination reagent;
[2][1]the homogeneous enzyme immunoassay method for the determination of 11-dehydro-thromboxane B as described in (1)2The reagent (2) consists of two parts (i.e. a first reagent R1 and a second reagent R2), characterized in that one of the reagents contains a labeled 11-dehydro-thromboxane B2The glucose-6-phosphate dehydrogenase of (1), correspondingly, another reagent contains 11-dehydro-thromboxane B2Antibody and substrate for glucose-6-phosphate dehydrogenase: oxidized Nicotinamide Adenine Dinucleotide (NAD)+) And glucose-6-phosphate (or a salt thereof);
[3] the urinary creatinine assay kit described in [1] consists of two parts (i.e. a third reagent R3 and a fourth reagent R4), and the components of the kit are consistent with those of a commercially available enzyme method creatinine determination reagent, namely R3 contains creatine hydrolase, sarcosine oxidase and catalase, and R4 contains creatinine hydrolase;
[4][1]the standard substance and the quality control substance of the aspirin drug resistance detection kit comprise creatinine and 11-dehydro-thromboxane B with certain concentration2The solution was mixed.
In particular, the use of the kit of the invention comprises 11-dehydro-thromboxane B2The determination and the creatinine determination are carried out on a full-automatic biochemical analyzer by using a double-channel four-reagent mode.
Wherein, 11-dehydro-thromboxane B2The detection is based on a homogeneous enzyme immunoassay, whichThe measurement principle is as follows: free 11-dehydro-thromboxane B in the sample2And 11-dehydro-thromboxane B labeled on glucose-6-phosphate dehydrogenase2Competitively bind 11-dehydro-thromboxane B2An antibody; when the antibody binds to glucose-6-phosphate dehydrogenase, the enzyme activity of glucose-6-phosphate dehydrogenase is greatly reduced, so that 11-dehydro-thromboxane B in the sample2The content is in direct proportion to the activity of the glucose-6-phosphate dehydrogenase in the detection system; by comparing with standard product, 11-dehydro-thromboxane B in the sample can be calculated2The content of (a).
The creatinine detection adopts an enzyme method, and the specific determination principle is as follows: creatine and sarcosine in the urine sample are eliminated by creatinase, sarcosine oxidase and catalase, and then sodium azide is used for inhibiting the activity of catalase in a system; then, creatinine reacts with creatininase, creatinase and sarcosine oxidase in sequence to generate hydrogen peroxide, the hydrogen peroxide is developed by utilizing Trinder's reaction, and the content of the creatinine in the sample can be calculated through comparison with a standard product.
Finally, by 11-dehydro-thromboxane B2And creatinine ratio to determine the patient's resistance to aspirin.
Drawings
FIG. 1 is a diagram of the correlation analysis of the kit.
Detailed Description
The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the present invention is not limited to these examples.
The reagents used in the examples are shown in table one.
Table one: sources of reagents in the examples
Figure 966478DEST_PATH_IMAGE001
Example 1: preparation of aspirin drug resistance detection kit
The aspirin drug resistance detection kit is prepared by the following method:
[1]preparing 11-dehydro-thromboxane B according to Table II2The reagent first, R1 in an aspirin resistance detection kit, was measured and its pH adjusted to 8.50.
Table two: 11-dehydro-thromboxane B2Assay reagent first reagent formulation
Figure 911431DEST_PATH_IMAGE002
[2]Preparing 11-dehydro-thromboxane B according to the third formula2The reagent second, R2 in an aspirin resistance detection kit, was measured and its pH adjusted to 8.50.
Table three: 11-dehydro-thromboxane B2Assay reagents second reagent formulations
Figure 922243DEST_PATH_IMAGE003
[3] A first reagent of the urine creatinine assay reagent, namely R3 in the aspirin resistance detection kit, is prepared according to the fourth table.
Table four: first reagent formula of urine creatinine determination reagent
Figure 474492DEST_PATH_IMAGE004
[4] And preparing a second reagent of the urine creatinine determination reagent, namely R4 in the aspirin resistance detection kit according to the fifth table.
Table five: second reagent formula of urine creatinine determination reagent
Figure 552563DEST_PATH_IMAGE005
The R1, R2, R3 and R4 prepared according to the scheme, a calibrator and a quality control product are combined to form the aspirin drug resistance detection kit.
Example 2: detection of aspirin drug resistance detection reagent and detection result
The performance of the aspirin resistance kit prepared in example 1 was used for detection according to the following procedure.
(1) Setting reaction parameters of a full-automatic biochemical analyzer: biochemical analyzer parameters of 11-dehydro-thromboxane B2 and creatinine were set according to table six and table seven, respectively.
TABLE VI.11-dehydro-thromboxane B2Biochemical instrument for measuring parameters
Figure 809363DEST_PATH_IMAGE006
TABLE seven. measurement parameters of urocreatinine biochemical instrument
Figure 307472DEST_PATH_IMAGE007
(2) Obtaining a standard curve
And (4) measuring the standard substance on a biochemical analyzer according to the parameters to obtain a standard curve.
(3) Sample detection
The standard curve obtained by the homogeneous enzyme immunoassay reagent of the invention is used for repeatedly measuring the high and low quality control samples for 10 times, and the detection data and the data analysis are shown in the table eight.
TABLE VIII reaction parameters of full-automatic biochemical analyzer
Figure 268606DEST_PATH_IMAGE008
And (3) detection results: the homogeneous enzyme immunoassay reagent has high accuracy and high precision, and CV is lower than 5%.
(4) Correlation analysis
The correlation analysis is carried out on 50 cases of clinical samples by respectively using a high performance liquid chromatography and the homogeneous phase enzyme immunoassay reagent of the invention, the measured data are shown in figure 1, and the obtained linear equation is as follows: y = 0.9838x +14.324 and the correlation coefficient R2 =0.9885, indicating that the detection reagent of the present invention determines 11-dehydrothromboxane B2The accuracy of clinical specimens is high.
It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and the scope of the invention is defined by the appended claims. Several modifications can be made without departing from the scope of the invention, and these modifications should also be construed as within the scope of the invention.

Claims (4)

1. In an aspirin drug resistance detection kit, the kit determines the drug resistance of a patient to aspirin by determining the ratio of 11-dehydro-thromboxane B2 to creatinine in human urine, and is characterized in that a homogeneous enzyme immunoassay method is used in an 11-dehydro-thromboxane B2 determination reagent of the kit, and the kit consists of a 11-dehydro-thromboxane B2 determination part, a creatinine determination part, a standard substance and a quality control substance.
2. The homogeneous enzyme immunoassay method of claim 1 for the determination of 11-dehydro-thromboxane B2The reagent (2) consists of two parts (i.e. a first reagent R1 and a second reagent R2), characterized in that one of the reagents contains a labeled 11-dehydro-thromboxane B2The glucose-6-phosphate dehydrogenase of (1), correspondingly, another reagent contains 11-dehydro-thromboxane B2Antibody and substrate for glucose-6-phosphate dehydrogenase: oxidized Nicotinamide Adenine Dinucleotide (NAD)+) And glucose-6-phosphate (or a salt thereof).
3. The urinary creatinine assay kit according to claim 1, which comprises two reagents (i.e., the third reagent R3 and the fourth reagent R4) and has a composition similar to that of a commercially available reagent for measuring creatinine by an enzymatic method, i.e., R3 contains creatine hydrolase, sarcosine oxidase and catalase, and R4 contains creatinine hydrolase.
4. The aspirin resistance test kit in claim 1, wherein the standard substance and quality control substance are creatinine and 11-dehydro-thromboxane B in certain concentration2The solution was mixed.
CN202010114209.6A 2020-02-25 2020-02-25 Aspirin drug resistance detection kit Pending CN111443201A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010114209.6A CN111443201A (en) 2020-02-25 2020-02-25 Aspirin drug resistance detection kit

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010114209.6A CN111443201A (en) 2020-02-25 2020-02-25 Aspirin drug resistance detection kit

Publications (1)

Publication Number Publication Date
CN111443201A true CN111443201A (en) 2020-07-24

Family

ID=71627149

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010114209.6A Pending CN111443201A (en) 2020-02-25 2020-02-25 Aspirin drug resistance detection kit

Country Status (1)

Country Link
CN (1) CN111443201A (en)

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6413459A (en) * 1987-07-08 1989-01-18 Ono Pharmaceutical Co Enzyme labeled thronboxane b2 and method for measuring 11-dehydro-thromboxane b2
EP0336647A2 (en) * 1988-03-31 1989-10-11 Sankyo Company Limited Determination of arachidonic acid derivatives and kits therefore
US20030124615A1 (en) * 1999-10-25 2003-07-03 Esoterix, Inc. Coagulation Thromoboxane B2 metabolite and methods for regulating aspirin-related platelet action
US20040115735A1 (en) * 2002-03-24 2004-06-17 Salim Yusuf Method for predicting cardiovascular events
US20070202556A1 (en) * 2005-12-09 2007-08-30 Corgenix Medical Corporation Methods and kits for detection of thromboxane a2 metabolites
CN102323414A (en) * 2011-06-07 2012-01-18 西安金域医学检验所有限公司 Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof
CN106124779A (en) * 2016-06-15 2016-11-16 四川迈克生物科技股份有限公司 A kind of test kit for measuring creatinine and method
CN106198509A (en) * 2016-06-15 2016-12-07 四川迈克生物科技股份有限公司 For measuring test kit and the method for creatinine
CN108627654A (en) * 2018-06-25 2018-10-09 武汉瀚海新酶生物科技有限公司 Composition for eliminating Calcium Dobesilate drug to the detection interference of creatinine enzyme process
CN109613226A (en) * 2019-01-24 2019-04-12 浙江夸克生物科技有限公司 A kind of creatinine assay kit and its application
CN111733208A (en) * 2020-08-15 2020-10-02 中山标佳生物科技有限公司 A kind of creatinine test kit capable of eliminating calcium dobesilate, sulfoethylamine and preparation method thereof

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS6413459A (en) * 1987-07-08 1989-01-18 Ono Pharmaceutical Co Enzyme labeled thronboxane b2 and method for measuring 11-dehydro-thromboxane b2
EP0336647A2 (en) * 1988-03-31 1989-10-11 Sankyo Company Limited Determination of arachidonic acid derivatives and kits therefore
US20030124615A1 (en) * 1999-10-25 2003-07-03 Esoterix, Inc. Coagulation Thromoboxane B2 metabolite and methods for regulating aspirin-related platelet action
US20040115735A1 (en) * 2002-03-24 2004-06-17 Salim Yusuf Method for predicting cardiovascular events
US20070202556A1 (en) * 2005-12-09 2007-08-30 Corgenix Medical Corporation Methods and kits for detection of thromboxane a2 metabolites
CN102323414A (en) * 2011-06-07 2012-01-18 西安金域医学检验所有限公司 Phenobarbital homogeneous-phase enzyme immunoassay reagent kit and preparation method thereof
CN106124779A (en) * 2016-06-15 2016-11-16 四川迈克生物科技股份有限公司 A kind of test kit for measuring creatinine and method
CN106198509A (en) * 2016-06-15 2016-12-07 四川迈克生物科技股份有限公司 For measuring test kit and the method for creatinine
CN108627654A (en) * 2018-06-25 2018-10-09 武汉瀚海新酶生物科技有限公司 Composition for eliminating Calcium Dobesilate drug to the detection interference of creatinine enzyme process
CN109613226A (en) * 2019-01-24 2019-04-12 浙江夸克生物科技有限公司 A kind of creatinine assay kit and its application
CN111733208A (en) * 2020-08-15 2020-10-02 中山标佳生物科技有限公司 A kind of creatinine test kit capable of eliminating calcium dobesilate, sulfoethylamine and preparation method thereof

Similar Documents

Publication Publication Date Title
Barsotti Measurement of ammonia in blood
Foster et al. A single-reagent manual method for directly determining urea nitrogen in serum
Bode et al. Spontaneous decay of oxidized ascorbic acid (dehydro-L-ascorbic acid) evaluated by high-pressure liquid chromatography
Lyman Blood urea nitrogen and creatinine
Witte et al. Evaluation of an enzymatic procedure for determination of serum cholesterol with the Abbott ABA-100
US6762035B1 (en) Method and test strips for the measurement of fat loss during weight loss programs
US6790668B1 (en) Monitoring patient compliance and bioavailability of drugs by deproteinizing body fluids
WO2022095576A1 (en) NOVEL N-ACETYL-β-D GLUCOSAMINIDASE DETECTION AGENT
CN102798726B (en) Cobalamin chemiluminescence immunoassay immue quantitative detection reagent box and preparation method thereof
CN112029817B (en) A creatinine detection kit and its use method
CN111289501A (en) Kit for detecting NO based on indirect colorimetric method and preparation and use methods thereof
JPS6091998A (en) Novel enzymatic measurement of d-3-hydroxybutyric acid and acetoacetic acid in humors, urine and measurement reagent for it
CN113655006B (en) Urinary system knot Dan Chengdan risk factor detection and test system
CN111443201A (en) Aspirin drug resistance detection kit
CN108359710A (en) The enzyme colour developing quantitative determination reagent kit and assay method of glucosephosphate isomerase
US4095948A (en) Determination of uric acid
CN108956971A (en) A kind of valproic acid immunologic function test reagent and its preparation and detection method
JP4217815B2 (en) Method for quantifying glucose by glucose dehydrogenase and reagent for quantifying glucose
JP2003502055A (en) A rapid and sensitive assay for homocysteine
CN108593905A (en) A digoxin immunoassay reagent and its preparation and detection method
Chen et al. Fluorescence polarization immunoassay and HPLC assays compared for measuring monoethylglycinexylidide in liver-transplant patients
Kiang et al. Semi-solid surface fluorometric analysis of glucose
CN1924580B (en) Chemiluminescent Assay Kit for Quantitative Detection of Triiodothyronine
CN113533577A (en) Method for simultaneously determining blood concentration of vancomycin and aminophylline and application
Okuda et al. Rapid and successive microdeterminations of oxygen, d-glucose, and its anomers in blood with β-d-glucose oxidase and oxygen electrode

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20200724