Summary of the invention
The object of the present invention is to provide a kind of assay method and equipment of platelet aggregation, this method is used a full-automatic platelet aggregation instrument, measure according to photoelectric turbidimetry, described gathering instrument comprises test machine core apparatus, cleaning disc device, reagent bottle device, sample disc device, sample needle and needle cleaning device; Full-automatic detection mode of the present invention; operating personnel have been avoided because of directly contacting contaminated with patient's blood sample; protected greatly that the operator's is healthy; all application of sample program, test procedure, cleaning procedure are finished automatically by instrument, have improved the test speed and the accuracy of clinical examination greatly.
An object of the present invention is to realize: a kind of assay method of platelet aggregation by following technical proposals, use a kind of full-automatic platelet aggregation instrument, measure according to photoelectric turbidimetry, described full-automatic platelet aggregation instrument comprises test machine core apparatus, cleaning disc device, reagent bottle device, sample disc device, sample needle and needle cleaning device, described sample needle is connected with automatic application of sample pipeline, and described cleaning needle is connected with automatic detergent line; Determination step is:
A, collection blood samples of patients sample carry out anti-freezing to this blood sample and handle, and generate anticoagulated whole blood, with the anticoagulated whole blood centrifugal treating, and preparation platelet rich plasma and platelet poor plasma;
B, the platelet aggregation derivant for preparing is put into the reagent bottle of full-automatic platelet aggregation instrument, cryopreservation is standby;
C, platelet rich plasma and the platelet poor plasma for preparing is placed in the sample disc device, sample number is set;
D, order sample needle are pressed the examination criteria consumption and are drawn platelet poor plasma from described sample disc device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads platelet poor plasma sample transmission data automatically; Clean sample needle and test cup then;
E, order sample needle are pressed the examination criteria consumption and are drawn platelet rich plasma from described sample disc device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads platelet rich plasma sample transmission data automatically; Clean sample needle and test cup then;
F, order sample needle are pressed the examination criteria consumption and are drawn the platelet aggregation derivant from the reagent bottle device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads the sample transmission data automatically; Automatically clean sample needle and test cup then;
The blood platelet MA of g, the tested blood sample of calculating generates test report according to the patient information of being imported simultaneously.
Another object of the present invention is realized by following technical proposals: a kind of full-automatic platelet aggregation instrument, a support is arranged, support is provided with photoelectric turbidimetry test circuit, automatic control circuit and matcher, it is characterized in that: set gradually test machine core apparatus, cleaning disc device, reagent bottle device, sample disc device on the testing stand of described support; Be provided with a plurality of test cups in the described test machine core apparatus, the detection light path system be set, comprise the lighting transistor circuit and the photoelectric switching circuit of complete configuration in this detection light path system around this test cup; Below the described test cup magnetic stirring apparatus is set; Comprise cleaning fluid bottle and waste collection bottle in the described cleaning disc device; On the described reagent bottle device low temperature holding device is set; On the described sample disc device a plurality of sample cups are set; Described testing stand top is provided with sample needle device and needle cleaning device, and sample needle and cleaning needle are installed on the running gear, this running gear comprise vertical travel mechanism, laterally travel mechanism, vertically to travel mechanism; Described sample needle device is connected with automatic application of sample pipeline, and described needle cleaning device is connected with automatic detergent line.
Compared with the prior art the present invention has following advantage:
1, adopts emission type test cup down in the test machine core apparatus of the present invention, make application of sample, detection, cleaning, removal waste fluid, dry entire exercise form the integral body of a robotization control.
2, the present invention has overcome the defective of prior art, and automatic application of sample, detection automatically, the automatic cleaning are incorporated on the instrument, has realized application of sample, detection, cleaning, removal waste fluid, dry full-automatic, has improved the test speed of clinical examination greatly.
3, full-automatic detection mode of the present invention makes operating personnel avoid having protected the healthy of operator greatly because of directly contacting contaminated risk with patient's blood sample, has simultaneously also avoided the operator pollution of blood sample to be influenced the accuracy of detection.
Embodiment
Embodiment one:
Referring to Fig. 1, the assay method of platelet aggregation of the present invention, use a kind of full-automatic platelet aggregation instrument, measure according to photoelectric turbidimetry, described full-automatic platelet aggregation instrument comprises test machine core apparatus, cleaning disc device, reagent bottle device, sample disc device, sample needle and needle cleaning device, described sample needle is connected with automatic application of sample pipeline, and described cleaning needle is connected with automatic detergent line; The technology contents of full-automatic platelet aggregation instrument is referring to the description of embodiment two, and determination step is:
A, collection blood samples of patients sample (pressing the collection of examination criteria consumption, normally 5ml) carry out anti-freezing to this blood sample and handle, and generate anticoagulated whole blood, with the anticoagulated whole blood centrifugal treating, and preparation platelet rich plasma and platelet poor plasma; In the present embodiment, the blood samples of patients sample carries out anti-freezing with blood anticoagulant 3.8% (concentration) sodium citrate, and the ratio of anti-coagulants and blood is 1: 9; Anticoagulated whole blood is injected sample hose, and to be moved into hydro-extractor then centrifugal, earlier with low-speed centrifugal (500~800 rev/mins, centrifugal 5 minutes), separate preparation PRP (being platelet rich plasma), with high speed centrifugation (3000 rev/mins, centrifugal 10 minutes), separate preparation PPP (being platelet poor plasma) again; In the present embodiment, also can adopt the index of control centrifugal force to realize separating, the present invention also can adopt LDI4-0.8 horizontal hydro-extractor (Beijing Medical Centrifugal Machine Factory);
B, the platelet aggregation derivant for preparing is put into the reagent bottle device of full-automatic platelet aggregation instrument, cryopreservation is standby; ADP more (adenosine diphosphate (ADP)) is generally used in the preparation of blood platelet derivant, and ADP generally is made into 3000umol/l (concentration) with physiological saline, and in packing in a small amount below-20 ℃, low temperature is frozen.Face with preceding and redissolve at room temperature, and be diluted to 300umol/l with physiological saline, put into the reagent bottle device of full-automatic platelet aggregation instrument then, it is standby to give stored refrigerated, refrigerated storage temperature 5-20 ℃; Derivant also can use a kind of among ADR (adrenaline), COL (collagen), ARA (arachidonic acid), the RIS (ristomycin).
C, platelet rich plasma and the platelet poor plasma for preparing is placed in the sample disc device, sample number is set; Set in advance corresponding calculated machine operation program in the full-automatic platelet aggregation instrument of the present invention, finished the setting of sample number by operating personnel's operational computations machine; Described sample number can be an electronic marker.
D, order sample needle are pressed the examination criteria consumption and are drawn platelet poor plasma from described sample disc device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads platelet poor plasma sample transmission data automatically; Clean sample needle and test cup then; In the present embodiment, the platelet poor plasma consumption is 300ul, described automatic cleaning sample needle and test cup, be according to pre-set program, use the clear water in the washer bottle to clean sample needle and test cup, sample needle is after each absorption sample is finished an application of sample action, and all the clear water that can introduce in the washer bottle cleans automatically, cleans the waste liquid that produces and is injected in the waste collection bottle of cleaning disc device by sample needle; After test action was finished, cleaning needle injected the cleaning of test cup with the clear water in the washer bottle, tested the discarded blood plasma in the cup and clean the vent valves of waste liquid bottom it to be discharged in the waste collection bottle; Start heating arrangement then to the oven dry of test cup;
E, order sample needle are pressed the examination criteria consumption and are drawn platelet rich plasma from described sample disc device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads platelet rich plasma sample transmission data automatically; Clean sample needle and test cup then; In the present embodiment, the platelet rich plasma consumption is 300ul, described automatic cleaning sample needle and test cup, be according to pre-set program, use the clear water in the washer bottle to clean sample needle and test cup, sample needle is after each absorption sample is finished an application of sample action, and all the clear water that can introduce in the washer bottle cleans automatically, cleans the waste liquid that produces and is injected in the waste collection bottle of cleaning disc device by sample needle; After test action was finished, cleaning needle injected the cleaning of test cup with the clear water in the washer bottle, tested the discarded blood plasma in the cup and clean the vent valves of waste liquid bottom it to be discharged in the waste collection bottle; Start heating arrangement then to the oven dry of test cup;
F, order sample needle are pressed the examination criteria consumption and are drawn the platelet aggregation derivant from the reagent bottle device, inject the test cup of test machine core apparatus, and the test machine core apparatus reads the sample transmission data automatically; Automatically clean sample needle and test cup then; In the present embodiment, platelet aggregation derivant consumption is 5ul, described automatic cleaning sample needle and test cup, be according to pre-set program, the cleaning fluid that sample needle is drawn in the cleaning fluid bottle cleans automatically, sample needle all can be drawn cleaning fluid and clean automatically after each absorption sample is finished an application of sample action, cleans the waste liquid that produces and is injected in the waste collection bottle of cleaning disc device by sample needle; After test action was finished, the cleaning fluid injection test cup that cleaning needle will be drawn in the cleaning fluid bottle cleaned, and the vent valves of waste liquid bottom it in the test cup is discharged in the waste collection bottle; Start heating arrangement then to the oven dry of test cup;
The blood platelet MA of g, the tested blood sample of calculating generates test report according to the patient information of being imported simultaneously; Described blood platelet MA account form is by four groups of transmittance-time-variable datas of full-automatic platelet aggregation instrument output, goes out the blood platelet MA according to these four groups of data computation then.The data of being measured are handled all belong to the proven technique content, be not described in detail at this.
In the present embodiment, the data test action of the action of the data test of platelet poor plasma, platelet rich plasma, the data test action of platelet aggregation derivant are undertaken by computer program control sequence.
Embodiment two:
Referring to Fig. 2, Fig. 3, Fig. 4, full-automatic platelet aggregation instrument of the present invention, a support is arranged, support is provided with photoelectric turbidimetry test circuit, automatic control circuit and matcher, sets gradually test machine core apparatus 7, cleaning disc device 6, reagent bottle device 5, sample disc device 4 on the testing stand 11 of described support 1; Be provided with a plurality of test cups 71 in the described test machine core apparatus, the detection light path system be set, comprise the lighting transistor circuit and the photoelectric switching circuit of complete configuration in this detection light path system around this test cup; Below the described test cup magnetic stirring apparatus is set; Comprise cleaning fluid bottle and waste collection bottle in the described cleaning disc device; On the described reagent bottle device low temperature holding device is set; A plurality of sample cups hole 41 is set on the described sample disc device, wherein places sample cup; Described testing stand top is provided with sample needle device and needle cleaning device 3, and sample needle 31 and cleaning needle 32 are installed on the running gear 2, this running gear comprise vertical travel mechanism 21, laterally travel mechanism 22, vertically to travel mechanism 23; Described sample needle device is connected with automatic application of sample pipeline, and described needle cleaning device is connected with automatic detergent line.
In the present embodiment, the photoelectric turbidimetry test circuit adopts prior art, is not described in detail.Computer Control Technology is adopted in control of the present invention, and its control circuit belongs to the routine techniques means, is not described in detail.
Referring to Fig. 3, in the present embodiment, be provided with 4 test cup positions in the test machine core apparatus, test cup 71 is plugged on the test cup position, can take out easily and insert, and magnetic stirring apparatus is set below the test cup, in the test cup, add a little bar magnet, magnetic agitation motor-driven disk rotates, and little bar magnet rotates at any time, realizes stirring the effect of plasma sample in the test cup; Belong to prior art, be not described in detail.The present invention measures according to photoelectric turbidimetry, the lighting transistor circuit and the photoelectric switching circuit that comprise complete configuration in the used detection light path system, comprise an infrarede emitting diode in the lighting transistor circuit, comprise an infrared receiving crystal pipe in the photoelectric switching circuit, cup one side of test cup is provided with infrarede emitting diode, and a corresponding survey is provided with infrared receiving crystal pipe (owing to being not show among the prior art figure).For test cup after cleaning is in time dried, below the test cup heating arrangement is set, this heating arrangement is a heating wiring board, generates heat by the lead on this wiring board, be used to reach the effect that makes test zone keep 37 ℃ of constant temperature, and the test cup after cleaning is in time dried.
Referring to Fig. 6, the test cup among the present invention is different with the employed square test cup of platelet aggregation instrument of the prior art, and the cup end of test cup of the prior art is sealed, and the present invention is open.In the present embodiment, the test cup comprises the transparent cup 711 of a pipe shape, and bottom of cup is provided with bottom outlet 712, installs back liquid pipe nipple 713 on the bottom outlet, and this liquid back pipe connects mouth and is connected with the liquid back pipe road.The present invention adopts pipe shape cup to be convenient to automatic cleaning, has eliminated the dead angle in the cup, improves cleaning degree; Adopt bottom outlet and liquid back pipe to connect mouth structure, can by control system realize the test cup externally by with the conversion of emissions status, make platelet aggregation instrument have the function of automatic washing, can be after having finished the blood sample test, raffinate is discharged in the waste collection bottle automatically, need not manually will test cup and extract and clean.
Referring to Fig. 2, Fig. 3, in the present embodiment, the top of cleaning disc device is provided with cleaning fluid bottle 61 and waste collection mouth 62, and the bottom is provided with the waste collection bottle, and the waste collection mouth is communicated with the waste collection bottle.Cleaning waste liquid that sample needle device, needle cleaning device, test cup produce and discarded blood plasma can be discharged in the waste collection bottle, the unified collection.
Referring to Fig. 2, Fig. 3, in the present embodiment, reagent bottle device top is provided with the reagent bottle hole, and its interpolation installed reagents bottle 51 is preserved the platelet aggregation derivant in the reagent bottle.In order to satisfy the temperature requirement of derivant, reagent bottle device bottom is provided with semiconductor cooling device.This refrigerating plant is made of refrigeration circuit, heat radiator and fan, and refrigeration circuit belongs to prior art, is not described in detail.
Referring to Fig. 2, Fig. 3, in the present embodiment, position, a plurality of sample cups hole 41 is set on the sample disc device, plug-in mounting has the sample cup of PRP, PPP in the sample cup hole, and the sample needle device can be drawn to detect from sample cup and use plasma sample.After detecting end, sample cup is discarded.
Referring to Fig. 2, Fig. 3, Fig. 4, in the present embodiment, sample needle in sample needle device and the needle cleaning device and cleaning needle are installed on the running gear 2, this running gear comprise vertical travel mechanism 21, laterally travel mechanism 22, vertically to travel mechanism 23; Vertically (X-direction) travel mechanism comprises longitudinal rail, stepper motor, synchronous cog belt, the carriage of vertically walking; Laterally (Y direction) travel mechanism comprises cross slide way, stepper motor, synchronous cog belt, the carriage of laterally walking; Vertically comprise vertically to guide rail, stepper motor, synchronous cog belt, vertically to the walking carriage to (Z-direction) travel mechanism; Laterally travel mechanism is installed on vertical walking carriage; Vertically be installed on the horizontal walking carriage to travel mechanism; Sample needle and cleaning needle are installed in vertically on the walking carriage.The sample needle device comprises sample needle, flexible pipe, solenoid valve, suction pumps, measuring element; Needle cleaning device cleaning needle, flexible pipe, solenoid valve, suction pumps, measuring element; This device belongs to the prior art scope, is not described in detail.
Referring to Fig. 5, the pipeline 8 of present embodiment comprises automatic application of sample pipeline and detergent line automatically.Sample needle 31 is connected with automatic application of sample pipeline, and this pipeline comprises solenoid valve, glass tube syringe pump 80, is used for drawing quantitative blood samples from sample cup, injects the test cup.The automatic detergent line of sample needle comprises washer bottle 86, T-valve 82, glass tube syringe pump, and deposit has clear water in the washer bottle, is used to clean sample needle.Needle cleaning device is connected with automatic detergent line, and the automatic detergent line of cleaning needle comprises washer bottle 86, T-valve 82, scavenging pump 81; Be used for cleaning the test cup.The liquid back pipe of test cup 71 connects mouth and is connected with the liquid back pipe road, and the liquid back pipe road comprises waste collection bottle 85, negative pressure pump 84, six-way valve 83, liquid level detection device, is used for test cup raffinate, discarded blood plasma, cleans waste collection.