Specific embodiment
Embodiment of the present invention is described in detail in conjunction with the embodiments, it will be appreciated by those skilled in the art that
The following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.Actual conditions are not specified in embodiment
Person, the condition suggested according to normal condition or manufacturer carry out.Reagents or instruments used without specified manufacturer, being can be with
Conventional products that are commercially available.
The present invention relates to a kind of kit of bleeding risk after detection thrombolysis, including serum creatinine detection reagent and leucocyte
Count reagent.
According to an aspect of the present invention, the invention further relates to serum creatinine detection reagents and white blood cell count(WBC) reagent to prepare
Application after detection thrombolysis in the kit of bleeding risk.
Preferably, as described above to apply, bleeding is bleeding after ST sections of elevation thrombolysis in myocardial infarction after the thrombolysis.
Current main creatinine assay method have chemical assay (alkaline picrate method), enzyme process, high performance liquid chromatography (HPLC),
Raman scattering method, isotope dilution mass spectrometry, capillary electrophoresis and electrode method etc..Isotope dilution mass spectrometry, Raman scattering
Method, high performance liquid chromatography (HPLC) are not suitable for the analysis of high-volume clinical samples, generally only as the reference method of creatinine assay, institute
Generally this method is not used to measure with clinical;Although capillary electrophoresis setting-out line range is wide, operation is relatively simple, needs to use
Special installation and the pretreatment for carrying out serum specimen, routine clinical use are more difficult;Alkaline picrate method and enzymatic assays creatinine
By clinical institute's extensive use.Alkaline picrate method has carried out alkaline picrate method various due to the interference of false creatinine, people
It improves, to improve the authenticity of measurement result, and realizes automated analysis, become clinically used detection method, but its line
Property range and antijamming capability are still not as good as enzyme process.Enzyme process kit interference resistance is stronger than picric acid method, and sample dosage is few, pollution
It is few, suitable for automatic clinical chemistry analyzer.Thus preferred, the serum creatinine detection reagent is enzyme process serum creatinine detection reagent.
Zymetology method is in the thrombolysis bleeding detection of the present invention, is had the characteristics that relatively stable and easily operated.
Preferably, the kit of bleeding risk or application, the serum creatinine detection reagent after detection thrombolysis as described above
Including creatine kinase, kreatinase, sarcosine oxidase and peroxidase.
By screening, the present invention measures creatinine level using zymetology method.Creatinine generates flesh under sarcosine oxidase effect
Propylhomoserin, sarcosine generate hydrogen peroxide under sarcosine oxidase effect, and the latter pacifies under hydrogen peroxide enzyme effect with 4- amino
For than woods, ESPAS reaction generation colour generation product aubergine quinone imines, absorbance is caused to increase at 545nm, pass through monitoring
The concentration risen to measure creatinine of absorbance value at 545nm.
Preferably, the kit of bleeding risk or application, the serum creatinine detection reagent after detection thrombolysis as described above
It further includes one or more in reaction buffer, color developing agent, cleaning agent, preservative:
Preferably, the reaction buffer is trihydroxy methyl amino buffer solution, Goods buffer solutions, glycine-NaOH are buffered
Liquid, N-2- hydroxyethyl piperazine-N'-2- ethanesulfonic acid buffers, N- tri- (methylol) methylamino -2- hydroxy-propanesulfonic acids buffer solution, N-
Bis- (2- hydroxyethanesulfonic acids) buffer solutions of three (methylol) methyl-2-amino ethanesulfonic acid buffers, piperazine-N, N-, 3- morpholine -2s-
Hydroxypropionate sodium buffer solution, 3- (N- morpholines) ethanesulfonic acid sodium buffer solution, 4- (2- ethoxys) piperazine -1-2- hydroxy-propanesulfonic acids
Bis- (2- ethoxys) amino -2- hydroxy-propanesulfonic acids bufferings of buffer solution, N- (2- ethoxys) piperazine-N'-4- fourths sulfonate buffer, 3-
Liquid, 3- (ring amine) -2- hydroxyl -1- propane sulfonic acid buffer solution, 4- (2- ethoxys) -1- piperazine propane sulfonic acid buffer solution, (ring is by 3-
Amine) -1- propane sulfonic acid buffer solution, 3- N-morpholinyls buffer solution, N- tri- (methylol) methyl-3-aminopropanesulfonicacid acid buffer solution one
Kind is several;
Preferably, the color developing agent is N- ethyls-N- (hydroxyl -3- sulfopropyls) meta-aminotoluene, potassium ferrocyanide and 4-
Amino-antipyrine;
Preferably, the preservative is potassium sorbate, in sodium benzoate, sodium nitrite, Proclin series preservatives
A kind of a kind of specific substance in specific substance or paraben esters;
It is furthermore preferred that the Proclin series preservative is Proclin300;
It is furthermore preferred that the paraben esters are methyl p-hydroxybenzoate, ethyl-para-hydroxybenzoate, para hydroxybenzene first
One kind in propyl propionate, butyl p-hydroxybenzoate, p-Hydroxybenzoic acid isopropyl ester, p-Hydroxybenzoic acid isobutyl ester.
Preferably, serum creatinine detection reagent of the present invention is divided into the first reagent and the second reagent;
First reagent includes creatine kinase, kreatinase;
Second reagent includes sarcosine oxidase, peroxidase, N- ethyls-N- (hydroxyl -3- sulfopropyls) meta-aminotoluene
(TOOS), 4-AA (PAP).
The detection sample that the serum creatinine detection reagent is detected can be serum or heparin blood plasma.
The stability of serum, blood plasma:4~25 DEG C of preservations can stablize 3 days.
Wherein, assay method principle is:
Absorption of the orchil of generation in 545nm is directly proportional to creatine concentration in sample.
Analysis type:End-point method deducts reagent blank;
Incubation time:5 minutes;
Reaction time:5 minutes;
The ratio of reagents of first reagent/sample/second:1800/60/600.Specifically used method is as shown in figure 3, absorbance becomes
The calculation formula of change is:
Δ A=[(A2-A1) calibrates quality control or sample cell]-[(A2-A1) blank tubes].
The computational methods of concentration are:
C samples=(Δ A samples/Δ A calibrations) × C calibrations;
C samples:Concentration of specimens for measure;
C standards:Concentration for calibration object.
In addition, the accuracy to further improve detection, can combine one or more detection reagents or tool for examining
The diagnosis of bleeding risk after thought-read vascular diseases, especially thrombolysis.
Current white blood cell count(WBC) is detected can be divided into capacitive, photoelectric type, laser class and electrical impedance according to principle difference
Type.Electrical impedance method detection leucocyte is Kurt (W.H.Coulter) invention by the U.S. earliest, is improved by decades, should
Testing principle is verified repeatedly in clinical practice.
By screening, present invention preferably employs electrical impedance method white blood cell count(WBC) reagents, have for Kurt theory of electrical impedance
Detect white blood cell count(WBC).According to Fig. 4, after quantitative blood is drawn and is diluted by quantitative conducting solution, just
It is sent to sensing chamber.In each sensing chamber, there are one small openings, are called detection aperture.Have in the both sides of detection aperture logical
There is the positive and negative electrode of constant current direct current.When diluted haemocyte by constant negative pressure act through detection aperture when, electricity
D.C. resistance between pole will change.This resistance can form a kind of proportional pulse of same blood cell volume size and become
Change.The distribution of particles that these data about pulse change being collected into can be used for one reflection size of blood cells of picture is bent
Line.
The pulse of each cell according to its volume size is distributed and is stored in corresponding when carrying out cell analysis by detector
Volume channel in, the data that each channel is collected are counted relative number, are represented in Y-axis, volume data is with ascend to heaven (fl)
For unit, represent in X-axis.Leucocyte volume can be divided from 30~450fl for 256 channels, each channel 1.64fl, foundation
Volume size is placed it in respectively in different channels, obtains the volume distributing histogram of leukocyte (as shown in Figure 5).
Preferably, the kit of bleeding risk or application, the white blood cell count(WBC) reagent after detection thrombolysis as described above
Ingredient include potassium dihydrogen phosphate, disodium hydrogen phosphate, cetyl trimethylammonium bromide, disodium ethylene diamine tetraacetate, ammonium oxalate
And glacial acetic acid.
It is furthermore preferred that the formula of the white blood cell count(WBC) reagent is:
Potassium dihydrogen phosphate:0.256~0.296g/L, disodium hydrogen phosphate:9.05~9.45g/L, disodium ethylene diamine tetraacetate:
15~25g/L, cetyl trimethylammonium bromide:7~8g/L, ammonium oxalate:15~25g/L, glacial acetic acid:20~30ml/L, it is molten
Agent is water.
Preferably, the kit of bleeding risk or application, the white blood cell count(WBC) reagent after detection thrombolysis as described above
Further include anti-coagulants;
Preferably, the anti-coagulants includes edta salt, heparinate, citrate, oxalates, hirudin, potassium fluoride, fluorination
It is one or more in sodium;
It is furthermore preferred that the edta salt is specially the potassium, sodium or lithium salts of EDTA;
It is furthermore preferred that the heparinate is specially the sodium, lithium or ammonium salt of heparin;
It is furthermore preferred that the citrate is specially the sodium or sylvite of citric acid;
It is furthermore preferred that the oxalates is specially the sodium or sylvite of oxalic acid.
In some embodiments, white blood cell count(WBC) can carry out in the following way:
1) reagent.Reagent includes diluent ingredient and hemolytic agent ingredient, configuration method:First match buffer solution, in a graduated cylinder,
Claim 4.6 grams of potassium dihydrogen phosphate, add 500 milliliters of distilled water, pH value 5.5, take 30 milliliters it is spare.In another graduated cylinder, claim phosphoric acid hydrogen
9.5 grams of disodium adds 1000 milliliters of distilled water, after dissolving, takes out 30 milliliters and does not have to, pH value is controlled 8.5, by two graduated cylinder solution
Mixing, pH value 8.5 claim 7.5 grams of cetyl trimethylammonium bromide, 20 grams of disodium ethylene diamine tetraacetate, 20 grams of ammonium oxalate, will be upper
It states buffer solution and adds to 1000 milliliters, 25 milliliters of ice acetic acid again after heating for dissolving are filtered, filtrate pH value 5.5 to 5.8 with filter paper
Between.The reagent of pre-configuration can be used for diluted blood cell, prevents blood cell aggregation and adhesion, for haemolysis red blood cell, beats
Broken red blood cell, in favor of white blood cell count(WBC).
2) sample requirement:Venous blood collection uses EDTA-K2 anti-freezings.Capillary blood sampling should wipe First Blood, so as not to it is mixed
Entering tissue fluid influences result.Collection of specimens is completed to measure after placing 5-10 minutes.
3) operating procedure:
A. small test tube 1 adds pre-configured leucocyte dilution+hemolytic agent 0.38ml.
B. 20 μ l of peripheral blood are accurately drawn with micropipet, wipe outside tip more than blood, will suction pipe be inserted into it is dilute in small test tube
The bottom of liquid is released, gently releases blood, and draws supernatant liquor, cleaning suction pipe is secondary, mixing.
C. pond is filled, 2~3min is stood, treats that leucocyte passes through respective aperture.
D. white blood cell count(WBC) is carried out with Counting software.
4) it calculates:It is quantified using the time, by the liquid volume in the unit interval is also constant when negative pressure is constant
, deviation that may be present is corrected using the counting for being carried out continuously same time twice, by the result that this is counted twice compared to pair,
It is quoted within the acceptable range as a result, carrying out third time counting if beyond normal acceptable range.Utilize software pair
Histogram is analyzed to judge.
According to an aspect of the present invention, the invention further relates to bleeding wind after a kind of ST sections of elevation thrombolysis in myocardial infarction of prediction
The method of danger, including:
For meeting the patient of thromboembolism treatment indication, obtained in patient's measuring samples using kit measurement as described above
Serum creatinine is horizontal and white blood cell count(WBC) data, transfer patient and symptom occur to consultation time, systolic pressure value and diabetes
Above-mentioned all data are distinguished assignment and calculate gross score by the data that history whether there is, and the gross score is total with pre-establishing
The correspondence of bleeding risk is compared to obtain bleeding risk knot after thrombolysis after-ST sections of elevation thrombolysis in myocardial infarction of score
Fruit.
Preferably, the measuring samples include whole blood, serum or blood plasma.
Preferably, the method that above-mentioned all data distinguish assignment is by method as described above:
The following institute of correspondence of bleeding risk after-ST sections of elevation thrombolysis in myocardial infarction of the gross score pre-established
Show:
According to an aspect of the present invention, the invention further relates to a kind of forecasting system of bleeding risk after thrombolysis, such as Fig. 1 institutes
Show, the system comprises:
Admission controller, serum creatinine detection part, the first assignment module, white blood cell count(WBC) component, the second assignment module, sample
This information receiving module, third assignment module and Risk Calculation module and reporting system main interface;
The admission controller is for assessing whether detected object meets thromboembolism treatment indication, will be by inspection pair if meeting
The sample to be checked of elephant is passed to serum creatinine detection part and the white blood cell count(WBC) component, and starts the sample information and receive work(
It can module;
The serum creatinine that the serum creatinine detection part is used to detect in the sample to be checked is horizontal, and serum creatinine is detected and is tied
Fruit is passed to the first assignment module and carries out assignment;
The white blood cell count(WBC) component is for will carry out white blood cell count(WBC) in the sample to be checked, and by white blood cell count(WBC) knot
Fruit is passed to the second assignment module and carries out assignment;
The sample information receiving module for receive detected object information and be passed to the third assignment module into
Row assignment;
The Risk Calculation module receives the first assignment module, the second assignment module and the third assignment
The assignment of module, and be calculated bleeding risk after thrombolysis according to the total score of each assignment;
The reporting system main interface is used to receive the sample information receiving module and/or the Risk Calculation mould
The information of block, and bleeding risk assessment result after thrombolysis is exported.
Wherein, thromboembolism treatment indication can be judged according to standard well known in the art, such as:Heart of Europe disease in 2017
Learn to provide in the ST-Elevation Acute Myocardial Infarction practice guidelines formulated:If in diagnosis ST-Elevation Acute Myocardial Infarction
It cannot row emergency PCI in time in 2 hours afterwards, it is proposed that the patient of no clear and definite contraindication is after symptom appearance
Thromboembolism treatment is carried out in 12 hours.
The serum creatinine detection part can be that serum creatinine detection reagent as described above and this field routine serum creatinine are examined
Survey pertinent instruments (particularly enzyme process serum creatinine detection pertinent instruments).
The white blood cell count(WBC) component can be white blood cell count(WBC) reagent as described above and this field conventional white cytometer
Number pertinent instruments (particularly electrical impedance method white blood cell count(WBC) pertinent instruments).
The system when in use, only can be only shown in client for serum creatinine detection part described in typing, described white
The interface of the output result of cell count component, and the interface also supports that being manually entered sample information receiving module is connect
The information (as shown in Figure 2) of receipts, and other component and/or module are hidden in backstage.After information is inputted, click " vertical
Calculate " button, you can it enters information into the Risk Calculation module, the main boundary of reporting system is then directly displayed at client
Face.
Preferably, after thrombolysis as described above bleeding risk forecasting system, controlled if the detected object does not meet thrombolysis
Indication is treated, then information is directly exported the reporting system main interface and generates suggestion intervention or drug is controlled by the admission controller
The report for the treatment of, while the system is closed by the admission controller.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the first assignment module passes through with lower section
Formula carries out assignment:
If 0 μm of ol/L≤88.4 μm of serum creatinine < ol/L, is scored at 0;
If 88.4 μm of ol/L≤176.8 μm of serum creatinine < ol/L, are scored at 5;
If serum creatinine >=176.8 are scored at 9.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the second assignment module passes through with lower section
Formula carries out assignment:
If white blood cell count(WBC)≤10 × 109, then it is scored at 0;
If white blood cell count(WBC)>10×109, then it is scored at 4.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the detected object information, which further includes, to be examined
, gender, weight, nationality, eating habit, life style, medication history, history of disease, there is symptom to medical at the age in the photo of object
One in time, household heredity factors, religious belief, heart condition, the Smoking And Drinking frequency, type of sports and the frequency, systolic pressure
It is or multinomial;Preferably, the history of disease is diabetic history;
Preferably, the detected object information includes at least systolic pressure, diabetic history and symptom occurs to consultation time.
Wherein, in the present invention, systolic pressure, diabetic history and occur symptom to consultation time definition be art technology
Well known to personnel, such as:
1) blood pressure measurement:A. patient takes clinostatism or seat, 5-10 minutes tranquil, and bilateral ancon is placed in heart level;B. it is
Patient's turn-up cuff dew arm (ancon stretches, and palmar is upward);C. sphygmomanometer is placed, is made at mercury " 0 " scale with arteria brachialis, at heart
In same level position, mercury cell switch is opened, drives residual air in sphygmomanometer girding to the greatest extent;D. it twines cuff is smooth in upper arm
Portion, lower edge are elastic to be advisable with being inserted into a finger away from fossa cubitalis 2-3CM;E. arteria brachialis is touched, wears stethoscope, stethoscope head is put in the upper arm
At arteriopalmus;F. valve screw-cap is closed, homogenous charge to auscultation brachial dance disappears, then increases 20-30mmHg.Slowly put
Gas, speed 4mmHg/s look squarely reading, and it is systolic pressure to hear the beating of the first sound, and beating is changed voice/disappeared as diastolic pressure.
2) diabetes medical history confirms:Diabetes of previously clarifying a diagnosis or the person that receives diabetes drug treatment;After being admitted to hospital on an empty stomach
Blood glucose >=7.0mmol/L (126mg/dl) or random plasma glucose >=11.1mmol/L (200mg/dl).
3) there is symptom to consultation time to confirm:There is symptom and be defined as before being this time admitted to hospital last breaking-out pectoralgia, uncomfortable in chest, dizzy
It faints, consultation time is defined as the assessment time started, and chronomere is accurate to hour.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the third assignment module passes through with lower section
Formula carries out assignment:
If systolic pressure < 90mmHg, are scored at 0;
If 90mmHg≤systolic pressure < 140mmHg, is scored at 3;
If 140mmHg≤systolic pressure < 180mmHg, is scored at 6;
If aglycosuria medical history, is scored at 0;
If there is diabetic history, it is scored at 4;
If 0h≤consultation time < 3h, is scored at 0;
If 3h≤consultation time < 6h, is scored at 2;
If consultation time >=6h, is scored at 4.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the Risk Calculation module is according to each assignment
Total score when be calculated bleeding risk after thrombolysis, the total score and the correspondence of bleeding risk after the thrombolysis
For:
Preferably, after thrombolysis as described above bleeding risk forecasting system, the system also includes system personnel permissions
Control module, control content include:
A) access right of each functional components of and/or module controls;
B) typings, audit, printing, the personnel's permission differentiation for cancelling audit;
C) defines artificial screen locking or locks screen automatically function without operation.
Preferably, after thrombolysis as described above bleeding risk forecasting system, the letter that the reporting system main interface is shown
Breath include one in following content, it is multinomial or whole:
1) calls the detected object information in the specimen information receiving module and is shown;
2) record of date informations and modification function;
The date information includes:Sampling time, the instrument detection date, user information date of entry, receives the sample presentation time
Instrumental results date, audit report date, printed report date and send reporting day interim one or more.
3) calls bleeding risk evaluation word template displaying after thrombolysis, and provides modification authority;
4) printing and establish self-defined report template that is reported;Custom item includes detected object number, report
Head, detected value, reference value, report picture, Health & Fitness Tip, auditor, printing people.
Preferably, after thrombolysis as described above bleeding risk forecasting system, bleeding risk is specially ST after the thrombolysis
Bleeding risk after section elevation thrombolysis in myocardial infarction.
Finally it should be noted that:The above embodiments are only used to illustrate the technical solution of the present invention., rather than its limitations;To the greatest extent
Pipe is described in detail the present invention with reference to foregoing embodiments, but it will be understood by those of ordinary skill in the art that:Its
It can still modify to the technical solution recorded in foregoing embodiments either to which part or all technical features
Carry out equivalent replacement;And these modifications or replacement, various embodiments of the present invention skill that it does not separate the essence of the corresponding technical solution
The range of art scheme.