CN104122404A - Novel rapid multi-parameter platelet functional analyzer and detection method thereof - Google Patents
Novel rapid multi-parameter platelet functional analyzer and detection method thereof Download PDFInfo
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- CN104122404A CN104122404A CN201410378146.XA CN201410378146A CN104122404A CN 104122404 A CN104122404 A CN 104122404A CN 201410378146 A CN201410378146 A CN 201410378146A CN 104122404 A CN104122404 A CN 104122404A
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Abstract
The invention discloses a novel rapid multi-parameter platelet functional analyzer. The novel rapid multi-parameter platelet functional analyzer comprises a multi-hole linked cup, a porous linked cup loading table, a diluting cup, a detection cup, a sample absorption needle, a sample absorption needle moving device, a diluent injector, a sample injector, a valve, an air pump, a waste liquid pump, a detection device, a control device and other main functional components. The invention also discloses a platelet functional analyzing method. The method comprises the following steps: the sample blood with equal quantity is respectively put into each hole of a special multi-hole linked cup; the multi-hole linked up filled with a blood sample is put onto the analyzer disclosed by the invention; the analyzer meters and detects the number and volume of platelets of the blood sample in each hole of the multi-hole linked cup, and finally compares and calculates the number of platelets in the blood sample in a contrast hole with the number of platelets in the blood sample in each other holes, thereby obtaining the aggregation ratio of the blood sample under action of different induced polymerization retarders respectively.
Description
Technical field
The present invention relates to a kind of novel platelet function analyser of multiparameter fast and detection method, is that a kind of novel can realizing carries out to platelet function in blood sample platelet function analytical instrument and the analytical approach thereof that multiparameter detects fast.
Background technology
Blood platelet is a kind of cell class particle composition in blood, is hemostasis and thrombotic key factor.Platelet function parameter is carried out to fast and easy detection, is an importance of platelet function research.That studies along with technical progress with to blood platelet gos deep into, and various countries have developed some instrument and methods to platelet function assay.As now existing: optics turbidimetry, electrode resistance method, thrombelastogram method, flow cytometer method etc.These methods or because complex operation, blood using amount are large, testing result is not good, somewhat expensive, detection speed be slow etc., and reason can not fine adaptation correlative study needs.
Using more platelet function assay instrument and method is at present photoelectric turbidimetry.But the method exists significantly not enough.Its operating process is: the whole blood of collection is first carried out to low-speed centrifugal, precipitation red blood cell, leucocyte wherein, supernatant after centrifugal is collected as platelet rich plasma, then wherein a part of platelet rich plasma is transferred in another centrifuge tube again, carrying out high speed centrifugation also precipitates down blood platelet wherein, collect supernatant blood plasma (being platelet poor plasma), then platelet counts in platelet rich plasma is adjusted, then first detect platelet poor plasma at special one on than turbid instrument, using this sample to instrument zeroing (as blank value), then detect platelet rich plasma (as blood platelet to light signal to obtained the maximum absorption), after detection, add induced polymerization inhibitor to detect platelet rich plasma and adding reduction (variation) curve of absorbance value after induced polymerization inhibitor, reduce and evaluate hematoblastic aggregation capability according to absorbance value.The method, due to large (generally each test needs 2ml whole blood) of its blood using amount, operates the defects such as very loaded down with trivial details, efficiency is low, testing result less stable.Therefore, at present platelet function assay instrument, method can't the corresponding scientific research of fine adaptation and clinical position needs.
Summary of the invention
Technical matters to be solved by this invention is for the deficiencies in the prior art, and a kind of platelet function analyser and detection method thereof of novel multiparameter is fast provided.
The technical solution that realizes the object of the invention is:
The invention provides a kind of novel platelet function of multiparameter fast analyser, described analyser comprises suction needle mobile device, porous connection cup microscope carrier, dilution and pick-up unit;
Porous connection cup microscope carrier comprises the porous connection cup in loading stage and loading stage, and porous connection cup is combined by the pipe of multiple (more than one) cuvette cartridge, and its inside is isolated from each other, and outside is connected to an entirety.
Dilution and pick-up unit comprise diluted cup, detection cup and pick-up unit, and diluted cup and detection cup are arranged side by side, and at detection cup sidewall, pick-up unit are set, and described pick-up unit is for detection of the quantity in blood platelet in blood sample to be measured.
Suction needle mobile device comprises suction needle and the device that drives suction needle to move, suction needle mobile device is for drawing the blood sample of porous connection cup, transfer them in the diluted cup in dilution and pick-up unit, and draw the blood sample after dilution in diluted cup, then transfer them to and detect in cup;
Wherein, porous connection cup microscope carrier and dilution and pick-up unit are arranged side by side, and suction needle mobile device is arranged on dilution and pick-up unit top.
Described analyser also comprises the first syringe, the second syringe, dilution memory storage, pneumatic pump and waste drains pump;
Wherein, suction needle connects the first syringe by pipeline, is realized absorption blood sample and is distributed blood sample by suction and the punching of the first syringe 10; Dilution memory storage is connected respectively diluted cup and is detected cup by pipeline, device for cleaning pipeline is to rim of a cup, and at rim of a cup inlet end, valve is set, between dilution memory storage and the pipeline of two glasss, be also provided with the second syringe, and between dilution memory storage and the second syringe, valve is set, for injecting dilution to two glasss.
Its course of work is as follows:
Open the valve between dilution memory storage and the second syringe, the second syringe extracts dilution in syringe pipeline, closes this valve, then opens the valve between the second syringe and diluted cup connecting pipe, promote the second syringe, the dilution in the second syringe injects diluted cup; If but now open the valve between the second syringe and analyzer cup connecting pipe, and promote the second syringe, the dilution in the second syringe injects analyzer cup.Cooperation between driving process and each valve of the second syringe is closed and is opened, and can realize by additional motor.
Waste drains pump is connected respectively diluted cup and is detected a cup cup end by pipeline, and diluted cup, detects cup and at cup bottom outlet end, valve is set respectively, and waste drains pump is connected with this pipeline, for the waste liquid in emptying glass; Between waste drains pump and two glasss of pipelines that are connected, be also provided with pneumatic pump, and at the endpiece of pneumatic pump, valve be set, for injecting an air to diluted cup and detection cup bottom, make blood sample and dilution fully dilute and mix.
Its course of work is as follows:
Open outlet of air pump end valve door, open at the same time or separately diluted cup and detect the valve on cup cup bottom outlet pipeline, pneumatic pump work can continuous delivery air in pipeline to two by cup bottom glass or in the open cup of valve, makes blood sample and dilution fully dilute and mix; Close the valve of pneumatic pump and outlet of air pump end, open diluted cup and detect a cup cup bottom outlet end valve door, after the detection of waste drains pump startup work in attracting two glasss, liquid is discharged outside instrument by pipeline under the driving of waste drains pump.Cooperation between above-mentioned valve is closed and is opened, and the control of waste drains pump and pneumatic pump is realized by equipment circuit and software.
In the present invention, described suction needle mobile device is used for driving suction needle to sample between porous connection cup, diluted cup, detection cup, the movement locus of suction needle is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, top, detection cup position.
Specific works process can realize in the following manner:
Described suction needle mobile device comprises suction needle, moves up and down unit, horizontal movement unit, move up and down unit and comprise two pulleys setting up and down, and travelling belt between pulley, girt is set between travelling belt, pull bar one end connects suction needle, two pulley driving travelling belts rotate, thereby drive girt vertical direction to move, and finally drive suction needle vertical direction to move.Horizontal movement unit comprises horizontally disposed two pulleys, and travelling belt between pulley, travelling belt with move up and down unit and be connected, two pulley driving travelling belts move reciprocatingly in the horizontal direction, thereby drive moves up and down unit horizontal direction and moves reciprocatingly, and finally drives suction needle to move reciprocatingly in the horizontal direction.Above-mentioned travelling belt moves up and down or moves horizontally all and can realize by additional motor.
In the present invention, described porous connection cup microscope carrier is used for loading porous connection cup, and to the blood sample to be measured in porous connection cup mix, stirring and/or heating and thermal insulation.
The invention provides a kind of implementation is: porous connection cup microscope carrier comprises the porous connection cup of loading stage, loading stage inside, two pulleys, and travelling belt between pulley; Wherein, two pulleys are setting up and down, and loading stage sidewall arranges top pulley, drive loading stage to rotate, and between two pulleys, connect by travelling belt.When work, the pulley rotation of drive motor drive below, thereby the upper square wheel rotation of drive, the porous in final drive loading cup joins the rotation of cup front and back, and rotational angle is no more than 85 °, thereby realizes the agitating function to each glass of interior blood sample.
The invention provides another kind of implementation is: in porous connection cup, in each glass, add bar magnet, in each glass of bottom of loading stage, stirring apparatus and magnet be set, thereby realize the agitating function to each glass of interior blood sample.
The loading stage inside of above-mentioned two kinds of structures is also provided with heating temperature control device, for the blood sample to be measured of porous connection cup is carried out to heated at constant temperature.
In the present invention, described porous connection cup is the aggregate of the pipe line spread of 3-8 test tube shape, and in porous connection cup, shape and the size of each glass are identical, and each glass of internal diameter suitable for reading is 8-25mm, each glass of endoporus clear depth of connection cup is 15-100mm, and the bottom in each glass of hole is semisphere or inverted-triangular.
In the present invention, the movement locus of described suction needle is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
The present invention also provides a kind of multiparameter platelet function analyser to detect the method for platelet counts in blood sample, comprises the following steps:
Step 1 does not add any induced polymerization inhibitor reagent cup in contrast respectively, or in this cup, packs in advance anti-coagulants into, and in other each glass, adds respectively the induced polymerization inhibitor of different cultivars or the induced polymerization inhibitor of same breed variable concentrations in one of them cup of porous connection cup;
Step 2, by same blood sample respectively equivalent join in each cup of porous connection cup, slight wobble mixes and blood sample and reagent is mixed and isothermal reaction at 22-37 DEG C;
Step 3, puts into porous connection cup microscope carrier by porous connection cup, constantly stirs and evenly mixs through porous connection cup microscope carrier;
Step 4, UNICOM's the first syringe and suction needle, draw the blood sample to be measured in contrast cup in porous connection cup, under the drive of suction needle mobile device, blood sample added in diluted cup, UNICOM's pneumatic pump and diluted cup, inject air and make blood sample and cup dilution fully mix and dilute;
Step 5, then the first syringe is crossed by UNICOM and suction needle is quantitatively drawn the blood sample in diluted cup, shifts to add and detects in cup, UNICOM's pneumatic pump and detection cup, inject air blood sample and cup dilution fully mixed and secondary dilution;
Step 6, the pick-up unit that is arranged on detection cup sidewall detects and record platelet counts, the volume of blood sample in contrast cup;
Step 7, after detection finishes, instrument cleans diluted cup, detection cup, sampling probe and each pipeline, UNICOM's waste drains pump and diluted cup, detection cup, emptying waste liquid; UNICOM's dilution memory storage and diluted cup, detection cup, savings dilution;
Step 8, blood sample repeating step 4～step 7 in other cup of distich cup, completes respectively other each glass of platelet counts, volume in porous connection cup is carried out to determination and analysis.
In the present invention, pick-up unit detects and record the platelet counts of blood sample in cup, the testing result of the platelet counts to each glass is calculated respectively and obtained to blood sample the platelet aggregation rate testing result under various different induced polymerization inhibitor effects according to following computing formula from contrast cup platelet counts, and computing formula is as follows:
In the present invention, pick-up unit detects and record the volume of platelets of blood sample in cup, utilize the size of the average external volume value that does not add induced polymerization inhibitor thromboblast is judged to whether blood sample is assembled before detection, thereby judge whether blood sample is assembled before detection, to avoid detecting mistake; Be greater than 12.5fl when above when not adding induced polymerization inhibitor thromboblast average external volume, pick-up unit is reported to the police, and prompting may be assembled by blood sample, thereby avoids the blood sample to there is platelet aggregation to detect the incorrect platelet aggregation rate result of acquisition.
The inventive method and instrument also detect hematoblastic volume in detecting platelet counts, and testing result represents with mean platelet volume (MPV).Instrument can be by detecting the size that does not add the hematoblastic average external volume of induced polymerization inhibitor blood sample (MPV) value, judge whether blood sample is assembled, its principle is if ought not adding induced polymerization inhibitor blood sample volume of platelets occurs obviously to strengthen (being greater than normal value peaked more than 20% or more than reaching 12.5fl), illustrates that obvious gathering has occurred blood sample.The blood sample that has occurred obviously to assemble will probably have larger difference with the aggregation rate value of normal blood sample, because utilizing the size of mean platelet volume (MPV) value that does not add induced polymerization inhibitor blood sample judged to whether blood sample is assembled before detection in detection method of the present invention, thereby help operator to find obviously to occur the blood sample of gathering, avoid detecting mistake.The present invention reports to the police not adding the blood sample that induced polymerization inhibitor blood sample mean platelet volume (MPV) is greater than more than 12.5fl, and prompting may be assembled by blood sample.
The inventive method and instrument can also be to not adding induced polymerization inhibitor sample, add induced polymerization inhibitor assembles the blood sample of platelet aggregation and respectively carries out twice or repeated detection, and use the mean value of testing result and the mean value of each gathering pattern detection result of not assembling platelet counts to compare respectively the aggregation rate of calculating the rear blood sample of the each induced polymerization inhibitor of acquisition (or variable concentrations induced polymerization inhibitor) effect, can improve like this stability of testing result.
The pick-up unit of the inventive method and instrument can be to detect wall of cup setting, also can be and detect glass independent agency separating, being connected the sample after dilution is transferred to independently and completed platelet count and the volume of platelets detection to diluting blood sample in pick-up unit by pipeline and detection cup.
Compared with prior art, its remarkable advantage is in the present invention:
(1) the suction needle mobile device in analyser of the present invention is used for driving suction needle freely to sample, distribute blood sample between each glass of porous connection cup, diluted cup, detection cup, automation mechanized operation, and efficiency improves greatly.
(2) connection of the porous in analyser of the present invention cup microscope carrier has the function of automatic mixing, automatic stirring and heated at constant temperature.
(3) dilution in analyser of the present invention and pick-up unit comprise diluted cup and detect cup for secondary dilution blood sample and carry out platelet counts and volume detection.
(4) setting of the injection of first in analyser of the present invention, the second syringe, dilution memory storage, pneumatic pump and waste drains pump, improve the sampling precision of sample and realized the vast scale dilution to sample, improved and detected the range of linearity of sample and the stability of testing result.
(5) the multi-joint cup in analyser of the present invention is combined by the pipe of multiple test tube shapes, its inside is isolated from each other, outside is connected to an entirety, its design is completely supporting with instrument, can realize a blood sample is carried out to multinomial induced polymerization inhibitor detection simultaneously, efficiency is much higher than existing detection method.
(6) in multiparameter platelet function analyser of the present invention and detection blood sample, the method for platelet counts has higher standardization and automatization level.
(7) described method can complete the detection that same blood examination is no less than to induced polymerization inhibitor that two kinds of induced polymerization inhibitor kinds or same breed the be no less than two kinds of variable concentrations sample to same blood sample processing at every turn simultaneously, can obtain the testing result of the platelet aggregation rate that is no less than two kinds of disparity items simultaneously.
(8) described method can be to not adding induced polymerization inhibitor sample, add induced polymerization inhibitor and assemble the blood sample of platelet aggregation sample and respectively carry out twice or repeated detection, and use the mean value of testing result of not assembling platelet counts and the mean value of each gathering pattern detection result to compare to calculate the aggregation rate result of blood sample after the each induced polymerization inhibitor effect of acquisition.
(9) the inventive method and instrument testing result can also be by original volume of platelets is provided, and help operator to judge whether blood sample is assembled before detection, thereby can avoid detecting mistake.
(10) analyser of the present invention detects the method for platelet counts and the coefficient R of turbidimetry Instrument measuring result in blood sample
2=0.8705, R=0.9330, correlativity is good, shows that the inventive method has higher feasibility.
Brief description of the drawings
Fig. 1 is analyser general structure schematic diagram of the present invention (1-suction needle mobile device, 2-porous connection cup microscope carrier, 3-dilution and pick-up unit).
Fig. 2 is the each parts connection diagram of analyser of the present invention.
Fig. 3 is suction needle mobile device structural representation of the present invention (9-suction needle, 15-moves up and down unit, 16-moves left and right unit, 17-cross bar).
Fig. 4 is the embodiment of the present invention 3 porous connection cup microscope carrier front views (4-porous connection cup, 5-loading stage, 18-top pulley, 19-below pulley, 20-motor).
Fig. 5 is the embodiment of the present invention 3 porous connection cup microscope carrier side views (4-porous connection cup, 5-loading stage, 18-top pulley, 19-below pulley).
Fig. 6 is the embodiment of the present invention 4 porous connection cup microscope carrier front views (4-porous connection cup, 21-stirring apparatus, 22-heating temperature control device, 23-bar magnet, 24-magnet).
Fig. 7 is porous connection cup stereographic map of the present invention.
To be suction needle of the present invention do to inhale to the blood sample in porous connection cup Fig. 8 fills blending manner schematic diagram.
Fig. 9 is that analyser of the present invention detects the aggregation rate of acquisition and the correlation analysis figure of optics turbidimetry Instrument measuring blood platelet MA.
Embodiment
Embodiment 1
As illustrated in fig. 1 and 2, a kind of novel platelet function of multiparameter fast analyser, described analyser comprises suction needle mobile device 1, porous connection cup microscope carrier 2, dilution and pick-up unit 3;
Porous connection cup microscope carrier 2 comprises the porous connection cup 4 in loading stage 5 and loading stage, and porous connection cup 4 is combined by the pipe of multiple test tube shapes, and its inside is isolated from each other, and outside is connected to an entirety;
Dilution and pick-up unit 3 comprise diluted cup 6, detection cup 7 and pick-up unit 8, and diluted cup 6 and detection cup 7 are arranged side by side, and at detection cup 7 sidewalls, pick-up unit 8 are set, and described pick-up unit 8 is for detection of the quantity in blood platelet in blood sample to be measured;
Suction needle mobile device 1 comprises suction needle 9 and the device that drives suction needle to move, suction needle mobile device is for drawing the blood sample of porous connection cup, transfer them in the diluted cup 6 in dilution and pick-up unit, and in diluted cup 6, draw the blood sample after dilution, then transfer them to and detect in cup 7;
Wherein, porous connection cup microscope carrier 2 is arranged side by side with dilution and pick-up unit 3, and suction needle mobile device 1 is arranged on dilution and pick-up unit 3 tops.
Described analyser also comprises the first syringe 10, the second syringe 11, dilution memory storage 12, pneumatic pump 13 and waste drains pump 14;
Wherein, suction needle 9 connects the first syringe 10 by pipeline, is realized absorption blood sample and is distributed blood sample (as shown in Figure 7) by suction and the punching of the first syringe 10; Dilution memory storage 12 is connected respectively diluted cup 6 and is detected cup 7 by pipeline, device for cleaning pipeline is to rim of a cup, and at rim of a cup inlet end, valve is set, between dilution memory storage 12 and the pipeline of two glasss, be also provided with the second syringe 11, and between dilution memory storage 12 and the second syringe 11, valve is set, for injecting dilution to two glasss;
At the bottom of waste drains pump 14 connects respectively diluted cup 6 by pipeline and detects 7 glasss, cup, and at cup bottom outlet end, valve is set, waste drains pump 14 endpiece arrange valve, for the waste liquid in emptying cup; Between waste drains pump 14 and the pipeline of two glasss, be also provided with pneumatic pump 13, and at the endpiece of pneumatic pump 13, valve be set, for injecting air to diluted cup 6 and detection cup 7 bottoms, make blood sample and dilution again fully dilute and mix.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 2
Other structure is with embodiment 1, as shown in Figure 3, suction needle mobile device comprises suction needle 9, moves up and down unit 15, horizontal movement unit 16, move up and down unit 15 and comprise two pulleys setting up and down, and travelling belt between pulley, girt 17 is set between travelling belt, pull bar one end connects suction needle 9, two pulley driving travelling belts rotate, thereby drive girt 17 vertical directions to move, and finally drive suction needle 9 vertical directions to move; Horizontal movement unit 16 comprises horizontally disposed two pulleys, and travelling belt between pulley, travelling belt with move up and down unit 15 and be connected, two pulley driving travelling belts move reciprocatingly in the horizontal direction, thereby drive moves up and down unit 15 horizontal directions and moves reciprocatingly, and finally drives suction needle 9 to move reciprocatingly in the horizontal direction.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 3
Other structure is with embodiment 1, and as shown in Figure 4, Figure 5, porous connection cup microscope carrier comprises the porous connection cup 4 of loading stage 5, loading stage 5 inside, two pulleys, and travelling belt between pulley; Wherein, two pulleys are setting up and down, and loading stage sidewall arranges top pulley 18, drive loading stage to rotate, and below pulley 19 drives rotation by drive motor 20, between two pulleys, connects by travelling belt; When work, drive motor 20 drives below pulleys 19 to rotate, thereby drives top pulley 18 to rotate, and rotate final porous connection cup 4 front and back of loading in cup that drive, and rotational angle is no more than 85 °, thereby realizes the agitating function to each glass interior blood sample.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 4
Other structure, with embodiment 1, as shown in Figure 6, adds bar magnet 23, in each glass of bottom of loading stage 5, stirring apparatus 21 and magnet 24 is set, thereby realize the agitating function to each glass of interior blood sample in porous connection cup in each glass.Loading stage 5 inside are provided with heating temperature control device 22.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 5
Other structure is with embodiment 2, and as shown in Figure 4, Figure 5, porous connection cup microscope carrier comprises the porous connection cup 4 of loading stage 5, loading stage 5 inside, two pulleys, and travelling belt between pulley; Wherein, two pulleys are setting up and down, and loading stage sidewall arranges top pulley 18, drive loading stage to rotate, and below pulley 19 drives rotation by drive motor 20, between two pulleys, connects by travelling belt; When work, drive motor 20 drives below pulleys 19 to rotate, thereby drives top pulley 18 to rotate, and rotate final porous connection cup 4 front and back of loading in cup that drive, and rotational angle is no more than 85 °, thereby realizes the agitating function to each glass interior blood sample.Loading stage 5 inside are provided with heating temperature control device 22.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 6
Other structure, with embodiment 2, as shown in Figure 6, adds bar magnet 23, in each glass of bottom of loading stage 5, stirring apparatus 21 and magnet 24 is set, thereby realize the agitating function to each glass of interior blood sample in porous connection cup in each glass.Loading stage 5 inside are provided with heating temperature control device 22.
In porous of the present invention connection cup, shape and the size of each glass are identical, are the aggregate of the pipe line spread of 3-8 test tube shape, and each rim of a cup internal diameter is 8-25mm scope, and the endoporus clear depth joining glass is 15-100mm, and the bottom in each glass of hole is semisphere.
The movement locus of suction needle of the present invention is rectilinear motion in the horizontal direction, does rectilinear movement sampling, distributes blood sample along each glass of porous connection cup, diluted cup, detection cup position.
Embodiment 7
Detect the method for platelet counts in blood sample based on above-mentioned multiparameter platelet function analyser, comprise the following steps: step 1, in porous connection cup 4 one of them cup, do not add any induced polymerization inhibitor reagent cup in contrast respectively, or in this cup, pack in advance anti-coagulants into (as sodium ethylene diamine tetracetate, EDTA), in other each glass, add respectively the induced polymerization inhibitor of different cultivars or the induced polymerization inhibitor of same breed variable concentrations;
Step 2, by same blood sample respectively equivalent join in porous connection cup 4 each cups, slight wobble mixes and blood sample and reagent is mixed and isothermal reaction at 22-37 DEG C;
Step 3, puts into porous connection cup microscope carrier 2 by porous connection cup 4, constantly stirs and evenly mixs through porous connection cup microscope carrier 2;
Step 4, UNICOM's the first syringe 10 and suction needle 9, draws the blood sample to be measured in contrast cup in porous connection cup 4, adds in diluted cup 6 under the drive of suction needle mobile device 1, UNICOM's pneumatic pump 13 and diluted cup 6, inject air and make blood sample and dilution fully mix and dilute;
Step 5, then UNICOM crosses the first syringe 10 and quantitatively draw the blood sample in diluted cup 6 with suction needle 9, adds and detects in cup 7, UNICOM's pneumatic pump 13 with detect cup 7, inject air blood sample and dilution fully mixed and secondary dilution;
Step 6, the pick-up unit 8 that is arranged on detection cup 7 sidewalls detects and record platelet counts, the volume of blood sample in contrast cup;
Step 7, after detection finishes, instrument cleans diluted cup, detection cup, sampling probe and each pipeline, UNICOM's waste drains pump 14 and diluted cup, detection glass, emptying waste liquid; UNICOM's dilution memory storage 12 and diluted cup, detection cup, savings dilution;
Step 8, blood sample repeating step 4～step 7 in other cup of distich cup, completes respectively other each glass of platelet counts, volume in porous connection cup is carried out to determination and analysis.
In the present invention, pick-up unit 8 detects and record the platelet counts of blood sample in cup, calculates respectively according to following formula detecting the platelet counts obtaining, and can obtain respectively the aggregation rate value of same blood sample under different induced polymerization inhibitor effects.
In the present invention, pick-up unit 8 detects and record the volume of platelets of blood sample in cup, utilize the size of the average external volume MPV-0 value that does not add induced polymerization inhibitor thromboblast is judged to whether blood sample is assembled before detection, thereby judge whether blood sample is assembled before detection, to avoid detecting mistake; Be greater than 12.5fl when above when not adding induced polymerization inhibitor thromboblast average external volume MPV, pick-up unit is reported to the police, and prompting may be assembled by blood sample.
Pick-up unit is to adopt Coulter principle (or being called theory of electrical impedance) to realize to the platelet counts in blood sample and volume detection method or implementation, and this technology is the ordinary skill in the art, and those skilled in the art are very familiar to clear.
Embodiment 8
Multiparameter platelet function analyser of the present invention and detection method test result repeatability, and detect the comparison of platelet aggregation rate results relevance with optics turbidimetry.
1) (this turbidimetry is referring to Born GVR.Aggregation of blood platelets by adenosine diphosphate and its reversal.Nature (Lond.) .1962 for instrument of the present invention and certain turbidimetry; 194:927) the repeatability comparison of Instrument measuring result.
Taking 1 part of sodium citrate anticoagulated whole blood as sample, detect platelet aggregation rate, each duplicate detection 10 times with instrument of the present invention and turbidimetry instrument respectively.The results are shown in following table.
Equipment in the present embodiment is identical with the equipment in embodiment 1, and step is identical with the step of embodiment 1, wherein, the dilution using is for going particulate etc. to ooze physiological saline, induced polymerization inhibitor is adenosine diphosphate (ADP) aqueous solution, and extension rate is 100 for the first time, and extension rate is 100 for the second time.
Table 1: the platelet aggregation rate result that instrument of the present invention and turbidimetry for Determination obtain
Try to achieve respectively the standard deviation that per minute is corresponding (SD) and mean value according to 10 measurement results.Press the row formula calculating coefficient of variation (CV) according to SD and mean value again.
CV=SD/ mean value × 100%, by calculating, the CV value of this instrument is 5%, and optics turbidimetry instrument gained is that my CV value is 11%, illustrates when this instrument is measured for aggregation rate, well repeated.
2) correlativity of instrument of the present invention and certain turbidimetry Instrument measuring result
Assemble instrument by this instrument and traditional turbidimetry and measure 40 increments originally simultaneously, two kinds of methods detect platelet aggregation rate to same sample respectively simultaneously, and 40 increments aggregation rate result is originally as follows:
Table 2: the result contrast of instrument of the present invention and turbidimetry Instrument measuring blood platelet MA
| Sample | Instrument of the present invention | Certain turbidimetry is assembled instrument |
| 1 | 75 | 61 |
| 2 | 56 | 50 |
| 3 | 54 | 48 |
| 4 | 66 | 59 |
| 5 | 45 | 36 |
| 6 | 69 | 61 |
| 7 | 77 | 63 |
| 8 | 48 | 40 |
| 9 | 49 | 43 |
| 10 | 78 | 62 |
| 11 | 65 | 58 |
| 12 | 51 | 40 |
| 13 | 49 | 34 |
| 14 | 62 | 49 |
| 15 | 51 | 44 |
| 16 | 65 | 53 |
| 17 | 54 | 43 |
| 18 | 46 | 42 |
| 19 | 78 | 62 |
| 20 | 68 | 54 |
| 21 | 43 | 33 |
| 22 | 65 | 59 |
| 23 | 69 | 55 |
| 24 | 39 | 30 |
| 25 | 45 | 40 |
| 26 | 57 | 45 |
| 27 | 49 | 39 |
| 28 | 46 | 39 |
| 29 | 54 | 49 |
| 30 | 34 | 25 |
| 31 | 63 | 57 |
| 32 | 68 | 59 |
| 33 | 54 | 47 |
| 34 | 53 | 39 |
| 35 | 62 | 44 |
| 36 | 51 | 37 |
| 37 | 65 | 48 |
| 38 | 59 | 53 |
| 39 | 58 | 52 |
| 40 | 46 | 42 |
Taking Instrument measuring result of the present invention as x axle, it is y axle that turbidimetry is assembled instrument measurement result, draws scatter diagram.Carry out linear regression analysis simultaneously, can obtain Fig. 9.From Fig. 9 result, the coefficient R of the inventive method and turbidimetry Instrument measuring result
2=0.8705, R=0.9330, correlativity is good.
But the result of Instrument measuring of the present invention is higher than than the result of turbidimetry Instrument measuring, two groups of data are matched to the result of T inspection and show, both have significant difference (P=0.000, P < 0.05).
The mode of the blood sample contrast that the present invention can process respectively by automatic detection platelet counts and with contrast cup blood sample and variant induced polymerization inhibitor quickly and easily, realizes multiple different induced polymerization inhibitor platelet aggregation rates is detected.And in detecting, can also obtain the information such as each detection platelet counts, mean platelet volume (MPV), erythrocyte number (RBC) and mean corpuscular volume (MCV).
The invention provides the platelet function analyser of novel multiparameter and the thinking of detection method.Method and the approach of this technical scheme of specific implementation are a lot; the above is only the preferred embodiment of the present invention; should be understood that; for those skilled in the art; under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.In the present embodiment not clear and definite each ingredient all available prior art realized.
Claims (10)
1. the novel platelet function of a multiparameter fast analyser, is characterized in that, described analyser comprises suction needle mobile device (1), porous connection cup microscope carrier (2), dilution and pick-up unit (3);
Suction needle mobile device (1) comprises suction needle (9) and drives the mobile device of suction needle (9), suction needle mobile device (1) is for drawing the blood sample of porous connection cup (4), transfer them in the diluted cup (6) in dilution and pick-up unit (3), and in diluted cup (6), draw the blood sample after dilution, then transfer them to and detect in cup (7);
Porous connection cup microscope carrier (2) comprises the porous connection cup (4) in loading stage (5) and loading stage, porous connection cup (4) is combined by the pipe of multiple test tube shapes, its inside is isolated from each other, and outside is connected to an entirety;
Dilution and pick-up unit (3) comprise diluted cup (6), detect cup (7) and pick-up unit (8), diluted cup (6) and detection cup (7) are arranged side by side, and at detection cup (7) sidewall, pick-up unit (8) is set, described pick-up unit (8) is for detection of the quantity in blood platelet in blood sample to be measured and volume;
Wherein, porous connection cup microscope carrier (2) is arranged side by side with dilution and pick-up unit (3), and suction needle mobile device (1) is arranged on dilution and pick-up unit (3) top.
Described analyser also comprises the first syringe (10), the second syringe (11), dilution memory storage (12), pneumatic pump (13) and waste drains pump (14);
Wherein, suction needle (9) connects the first syringe (10) by pipeline, is realized absorption blood sample and is distributed blood sample by suction and the punching of the first syringe (10); Dilution memory storage (12) is connected respectively diluted cup (6) and is detected cup (7) by pipeline, device for cleaning pipeline is suitable for reading to diluted cup and detection cup, and approach cup-mouth end at pipeline valve is set, between dilution memory storage (12) and the pipeline of two glasss, be also provided with the second syringe (11), and between dilution memory storage (12) and the second syringe (11), valve is set, for injecting dilution to two glasss;
Waste drains pump (14) is connected respectively diluted cup (6) and is detected cup (7) the cup end by pipeline, and at cup bottom outlet end, valve is set, and waste drains pump (14) endpiece arranges valve, for the waste liquid in emptying cup; Between waste drains pump (14) and the pipeline of two glasss, be also provided with pneumatic pump (13), and the endpiece at pneumatic pump (13) arranges valve, for injecting an air to diluted cup (6) and detection cup (7) bottom, make blood sample and dilution fully dilute respectively and mix.
2. multiparameter platelet function analyser according to claim 1, it is characterized in that, suction needle mobile device (1) comprises suction needle (9), move up and down unit (15), horizontal movement unit (16), move up and down unit (15) and comprise two pulleys setting up and down, and travelling belt between pulley, girt (17) is set between travelling belt, pull bar one end connects suction needle (9), two pulley driving travelling belts rotate, thereby drive girt (17) vertical direction to move, final suction needle (9) vertical direction that drives moves, horizontal movement unit (16) comprises horizontally disposed two pulleys, and travelling belt between pulley, travelling belt with move up and down unit (15) and be connected, two pulley driving travelling belts move reciprocatingly in the horizontal direction, thereby drive moves up and down unit (15) horizontal direction and moves reciprocatingly, the final suction needle (9) that drives moves reciprocatingly in the horizontal direction, finally realizes suction needle and carries out arbitrarily to-and-fro movements upper and lower, front and back.
3. multiparameter platelet function analyser according to claim 1, it is characterized in that, porous connection cup microscope carrier (2) comprises porous connection cup (4), two pulleys of loading stage (5), loading stage inside, and travelling belt between pulley; Wherein, two pulleys are setting up and down, and loading stage sidewall arranges top pulley (18), drive loading stage (5) to rotate, below pulley (19) is driven and is rotated by drive motor (20), between two pulleys, connects by travelling belt; When work, drive motor (20) drives below pulley (19) to rotate, thereby drive top pulley (18) to rotate, before and after the final porous connection cup (4) driving in loading stage (5), rotate, its rotational angle is that any side is no more than 85 °, thereby realizes the agitating function to each glass of interior blood sample.
4. multiparameter platelet function analyser according to claim 1, it is characterized in that, in porous connection cup, in each glass, add bar magnet (23), in each glass of bottom of loading stage (5), stirring apparatus (21) and magnet (24) be set, thereby realize the agitating function to each glass of interior blood sample.
5. according to the arbitrary described multiparameter platelet function analyser of claim 1～4, it is characterized in that, loading stage inside is provided with heating temperature control device (22).
6. according to the arbitrary described multiparameter platelet function analyser of claim 1～4, it is characterized in that, described porous connection cup (4) is the aggregate of the pipe line spread of 3-8 test tube shape, in porous connection cup, shape and the size of each glass are identical, each glass of internal diameter suitable for reading is 8-25mm, each glass of endoporus clear depth of connection cup is 15-100mm, and the bottom in each glass of hole is semisphere.
7. according to arbitrary described multiparameter platelet function analyser in claim 1～4, it is characterized in that, the movement locus of described suction needle is rectilinear motion in the horizontal direction, and the position distributing along each glass of porous connection cup, diluted cup, detection rim of a cup is done the sampling that moved linearly, distribution, shifted blood sample.
8. the method that detects platelet counts in blood sample based on arbitrary described multiparameter platelet function analyser in claim 1～4, is characterized in that, comprises the following steps:
Step 1, in one of them cup of porous connection cup (4), do not add respectively any induced polymerization inhibitor reagent cup in contrast, or in this cup, pack in advance anti-coagulants into, and in other each glass, add respectively the induced polymerization inhibitor of different cultivars or the induced polymerization inhibitor of same breed variable concentrations;
Step 2, by same blood sample respectively equivalent join in each cup of porous connection cup (4), slight wobble mixes and blood sample and reagent is mixed and isothermal reaction at 22-37 DEG C;
Step 3, puts into porous connection cup microscope carrier (2) by porous connection cup (4), constantly stirs and evenly mixs through porous connection cup microscope carrier (2);
Step 4, UNICOM's the first syringe (10) and suction needle (9), draw the blood sample to be measured in contrast cup in porous connection cup (4), under the drive of suction needle mobile device (1), blood sample is added in diluted cup (6), UNICOM's pneumatic pump (13) and diluted cup (6), inject air and make blood sample and cup dilution fully mix and dilute;
Step 5, UNICOM crosses the first syringe (10) and quantitatively draws the blood sample in diluted cup (6) with suction needle (9) again, transfer joins and detects in cup (7), UNICOM's pneumatic pump (13) and detection cup (7), inject air blood sample and cup dilution fully mixed and secondary dilution;
Step 6, the pick-up unit (8) that is arranged on detection cup (7) sidewall detects and record platelet counts, the volume of blood sample in contrast cup;
Step 7, after detection finishes, instrument cleans diluted cup, detection cup, sampling probe and each pipeline, UNICOM's waste drains pump (14) and diluted cup, detection glass, emptying waste liquid; UNICOM's dilution memory storage (12) and diluted cup, detection cup, savings dilution;
Step 8, blood sample repeating step 4～step 7 in other cup of distich cup, completes respectively other each glass of platelet counts, volume in porous connection cup is carried out to determination and analysis.
9. the method for platelet counts in detection blood sample according to claim 8, it is characterized in that, pick-up unit (8) detects and record the platelet counts of blood sample in cup, calculate and obtain the aggregation rate of blood sample under various induced polymerization inhibitor effects according to following calculating respectively according to detecting the platelet counts obtaining in the cup of each hole, its computing formula is as follows:
10. the detection method of platelet aggregation in detection blood sample according to claim 8, it is characterized in that, pick-up unit (8) detects and record the volume of platelets of blood sample in cup, utilize the size of the average external volume value that does not add induced polymerization inhibitor thromboblast is judged to whether blood sample is assembled before detection, thereby judge whether blood sample is assembled before detection, to avoid detecting mistake; Be greater than 12.5fl when above when not adding induced polymerization inhibitor thromboblast average external volume, pick-up unit is reported to the police, and prompting may be assembled by blood sample.
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| CN105699434A (en) * | 2016-01-28 | 2016-06-22 | 陈晓乾 | Method and device for measuring content of blood platelets |
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| CN107966580A (en) * | 2017-12-26 | 2018-04-27 | 江苏柯伦迪医疗技术有限公司 | A kind of fast automatic biochemical detection system and method for application bar formula detection card |
| CN108627637A (en) * | 2018-06-04 | 2018-10-09 | 江苏柯伦迪医疗技术有限公司 | A kind of platelet aggregation detecting system and method |
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| CN112526144A (en) * | 2020-12-29 | 2021-03-19 | 滁州瑞谷生物技术有限公司 | Platelet aggregation rate detection method and detection device |
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