CN103344638B - A kind of semi-automatic external adopting colorimetry to measure HCY detection equipment - Google Patents
A kind of semi-automatic external adopting colorimetry to measure HCY detection equipment Download PDFInfo
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- CN103344638B CN103344638B CN201310281930.4A CN201310281930A CN103344638B CN 103344638 B CN103344638 B CN 103344638B CN 201310281930 A CN201310281930 A CN 201310281930A CN 103344638 B CN103344638 B CN 103344638B
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Abstract
The invention discloses a kind of semi-automatic external adopting colorimetry to measure HCY detection Apparatus and method for, described method includes: when the first detectable and the detected sample response time in cuvette being detected then, microprocessor controls sample needle moves to cuvette place, and is directed at cuvette center；Microcontroller start-up loading pump, draws the second detectable from the container holding the second detectable, and adds it to react in cuvette；The content of HCY in detected sample is determined by the absorbance of reacting substance in cuvette.Present invention achieves the automatic loading of the second detectable, eliminate the artificial operation loading the second detectable, not only save the cost of labor of HCY detection, but also testing result is impacted by the operation error being avoided as much as artificial dropping detectable；The present invention improves the detection efficiency of HCY simultaneously, substantially reduces the detection time of HCY, has wide market prospect.
Description
Technical field
The present invention relates to in-vitro diagnosis equipment, in particular a kind of semi-automatic external adopting colorimetry to measure HCY detection Apparatus and method for.
Background technology
The aminoacid of a kind of sulfur-bearing that homocysteine (homocysteine, HCY) Methionine metabolism produces, the HCY of 80% is by disulfide bond and protein binding in blood, and only seldom circulation participated in by the free homocysteine of a part.HCY level is closely related with cardiovascular disease, is a significant risk factor of cardiovascular disease incidence, and the Hcy increased in blood, because stimulating blood vessel wall to cause arterial vascular damage, causes that the speckle of inflammation and tube wall is formed, finally causes cardiac flow to be obstructed.Vitamin B malnutrition can raise with moderate or slight HCY, also can increase cardiopathic danger.In recent years, domestic and international many scholars all think that blood homocysteine content raises has become the independent risk factor that atherosclerosis occurs, then becomes the new focus of preclinical medicine and clinic study.
At present, the common methods measuring HCY is Enzymatic cycling, and the method is first to be added in detectable R1 by detected sample, reaction a period of time adds detectable R2, after reacting a period of time again, measure the change of the absorbance of product, determine the content of HCY in detected sample.
The existing equipment based on colorimetric determination HCY has two kinds, and one is large automatic biochemistry analyzer, is generally used for clinical laboratory or the central laboratory of big-and-middle-sized hospital, another kind of small semiautomatic detection equipment, is commonly used in clinical department or She Kang center.
Current small semiautomatic HCY detects equipment as shown in Figure 1, monitor station 10 is provided with multiple Photoelectric Detection module 20 with temperature control and agitating function, during measurement, detected sample S is manually adding to be pre-loaded with in the cuvette 21 of detectable R1 by testing staff, again cuvette 21 is inserted in Photoelectric Detection module 20, two kinds of materials are made to react a period of time in cuvette, then again detectable R2 is added in cuvette 21, react a period of time again, the absorbance of product in cuvette is measured by the microcontroller of detection equipment, thus calculating the content of HCY in detected sample.But this detection mode also exists following defect:
1) detectable R2 need to add manually, owing to cuvette is upper end tapered opening, lower end is chamber, Vertical Narrow footpath, during artificial instillation detectable, very easily due to the operational error of testing staff, dropping on the inwall of cuvette by part detectable, the amount making the detectable that participation reacts is not enough, and then affects testing result；
2) reaction needed a period of time of detectable R1 and detected sample, when instrument prompting operator add detectable R2, owing to the skilled operation degree of each operator or the speed of addition detectable can be different, therefore, this can make detectable exist different from the response time of detected sample, causes and also can there are differences between testing result；Meanwhile, the mode of this artificial interpolation adds the workload of testing staff, and easily occurs that monitoring error affects testing result.
3) addition of detectable R2 is usually a few microlitre, and operator have significantly high skill set requirements, and careless slightly to will result in detectable addition not enough, very easily produces error, thus affecting the accuracy of measurement result.
Therefore, prior art has yet to be improved and developed.
Summary of the invention
For the drawbacks described above of prior art, the present invention provides a kind of semi-automatic external adopting colorimetry to measure HCY detection Apparatus and method for, it is achieved the automatic loading of detectable, and then realizes the automatization of HCY detection.
This invention address that the technical scheme that technical problem adopts is as follows:
A kind of semi-automatic external adopting colorimetry to measure HCY detection equipment, including a monitor station, described monitor station is provided with the Photoelectric Detection module of temperature control and agitating function, described Photoelectric Detection module includes an optical filter, described Photoelectric Detection module is placed with the cuvette pre-installing the first detectable, described monitor station is additionally provided with a mechanical arm, load pump and hold the container of the second detectable, described mechanical arm is provided with a sample needle, one end of described load pump connects described sample needle, the other end connects described container, monitor station is additionally provided with the microcontroller for controlling described detection device detection procedure.
Described employing colorimetry measures the semi-automatic external detection equipment of HCY, and wherein, described Photoelectric Detection module includes several.
Described employing colorimetry measures the semi-automatic external detection equipment of HCY, wherein, described Photoelectric Detection module on monitor station linearly or circular arc arrangement.
Described employing colorimetry measures the semi-automatic external detection equipment of HCY, and wherein, described load pump is dosing pump.
Described employing colorimetry measures the semi-automatic external detection equipment of HCY, and wherein, described dosing pump is plunger displacement pump or syringe pump.
Described employing colorimetry measures the semi-automatic external detection equipment of HCY, and wherein, described Photoelectric Detection module includes one being arranged on the light source of cuvette side, being arranged on cuvette opposite side with described light source just to the photoelectric sensor arranged.
Colorimetry is adopted to measure the semi-automatic external detection method of HCY according to described detection equipment, wherein, including step:
When the first detectable and the detected sample response time in cuvette being detected then, microprocessor controls sample needle moves to cuvette place along mechanical arm, and is directed at cuvette center；
Microcontroller start-up loading pump, draws the second detectable from the container holding the second detectable, and adds it to react in cuvette；
The content of HCY in detected sample is determined by the absorbance of reacting substance in cuvette.
Described employing colorimetry measures the semi-automatic external detection method of HCY, wherein, determined in detected sample by the absorbance of reacting substance in cuvette and also include after the step of the content of HCY: microcontroller again controls sample needle and moves, sample needle is moved in next cuvette, and the second detectable is joined in this cuvette, carry out next group detection.
Employing colorimetry provided by the present invention measures the semi-automatic external detection Apparatus and method for of HCY, its technical advantages reached is: by the setting of mechanical arm, load pump and sample needle, realize the automatic loading of detectable, save the cost of labor of HCY detection, it is to avoid testing result is impacted by the operation error of artificial dropping detectable；The present invention improves the detection efficiency of HCY simultaneously, has wide market prospect.
Accompanying drawing explanation
Fig. 1 is the structural representation that in prior art, HCY detects equipment.
Fig. 2 is the structural representation that employing colorimetry provided by the invention measures the semi-automatic external detection equipment of HCY.
Fig. 3 is that employing colorimetry provided by the invention measures the structural representation of cuvette in the semi-automatic external detection equipment of HCY.
Fig. 4 is that employing colorimetry provided by the invention measures the structural representation of Photoelectric Detection module in the semi-automatic external detection equipment of HCY.
Fig. 5 is the flow chart that employing colorimetry provided by the invention measures the semi-automatic external detection method of HCY.
Fig. 6 is the flow chart that employing colorimetry provided by the invention measures the semi-automatic external detection method another preferred embodiment of HCY.
Detailed description of the invention
For making the purpose of the present invention, technical scheme and advantage clearly, clearly, developing simultaneously referring to accompanying drawing, the present invention is described in more detail for embodiment.Should be appreciated that specific embodiment described herein is only in order to explain the present invention, is not intended to limit the present invention.
The invention provides a kind of semi-automatic external adopting colorimetry to measure HCY detection equipment, as shown in Figure 2, including a monitor station 100, described monitor station 100 is provided with several Photoelectric Detection module 110 with temperature control and agitating function, and on these Photoelectric Detection modules 110, it is placed with cuvette 200 respectively, cuvette 200 is used for pre-installing the first detectable and as the first detectable, the reaction vessel of detected sample and the second detectable, described monitor station 100 is additionally provided with a mechanical arm 101, mechanical arm 101 is fixed on monitor station 100 by screw 140, mechanical arm 101 is also equipped with a sample needle 103, sample needle can move along mechanical arm 101, monitor station is also provided with a load pump 102, one end of this load pump 102 connects described sample needle 103, its other end is connected in the container 104 being contained with the second detectable, this container 104 is arranged on monitor station 100, load pump 102 is connected by conduit 120 with sample needle 103 and container 104, but it is not limited to conduit.
Further, vitro detection equipment provided by the invention can detect for single channel, it can be multi-channel detection, during single channel detection, one Photoelectric Detection module is only set on monitor station, and during multi-channel detection, the present embodiment includes several Photoelectric Detection module, and now Photoelectric Detection module is arranged in a linear on monitor station, the quantity of Photoelectric Detection module is arranged as required, being typically but not limited to 4 to 8, the modes of emplacement of this straight-line has advantage clearly relative to existing circular modes of emplacement.Cuvette structural representation as shown in Figure 3, cuvette 200 is generally upper end pyramidal structure, opening is square, and lower end is vertical cavity, and vertical cavity diameter is less, these cuvettes are placed on Photoelectric Detection module, if adopting circle to place, then the placement orientation of each cuvette is different, in the process instilling detectable or detected sample wherein one by one, easily cause detectable or detected sample and, on the inwall dropping in cuvette, affected testing result.And adopt straight-line that Photoelectric Detection module is set and can reduce the probability that detectable or detected sample are dropped on cuvette inwall as much as possible, reduce error risk.Meanwhile, straight-line arranges Photoelectric Detection module and also can be greatly simplified the control of sample needle.Certainly, Photoelectric Detection module is not limited to adopt linear array on monitor station, it would however also be possible to employ circular arc arrangement or other arrangement modes.
The structure of the mechanical arm of the present invention is adaptive with the arrangement mode of Photoelectric Detection module, in the present embodiment, owing to Photoelectric Detection module is linear arrangement, therefore the mechanical arm of the present invention is preferably linear structure, and this linear structure can simplify sample needle movement on the robotic arm.Mechanical arm can be arranged on but be not limited to the surface of Photoelectric Detection module, it is preferred to surface, so, sample needle just can be directed at cuvette, it is simple to the interpolation of detectable.
In order to well realize the automatic loading of the second detectable, the present invention can arrange a microcontroller 130 with timing function on this vitro detection equipment, this microcontroller 130 is used for controlling this detection device detection procedure, moves on the robotic arm and controlled loading pumping takes the second detectable specifically for automatically controlling sample needle；The response time of the first detectable Yu detected sample can also be timed simultaneously, and can the response time then, control sample needle and move along mechanical arm, simultaneously start-up loading pump, draw the second detectable from container, and by sample needle, the second detectable is instilled in cuvette.
Specifically, second detectable is held by special container 104, this container 104 is arranged on monitor station 100, concrete placement location does not limit at this, owing to load pump connects this container 104 by conduit, therefore, this container 104 freely can be arranged on monitor station, is certainly preferably the suitable position manually picked and placeed.Carrying out when automatically loading of the second detectable, first, microprocessor controls sample needle moves and moves along mechanical arm, and final positive contrast color cup；Secondly, microcontroller start-up loading pump, draw appropriate second detectable from container, then pass through sample needle and be added dropwise in cuvette.The present embodiment passes through microprocessor controls load pump, controls the uptake of the second detectable, in order to testing staff needs to adjust according to detection.
Depositing simultaneously as the second detectable need to be freezed, therefore, the second container of the present embodiment is preferably and is positioned in refrigerating plant.
The load pump of the present invention is dosing pump, for instance plunger displacement pump or syringe pump, specifically can adopt the high accuracy injection measurement pump of valveless plunger displacement pump or three-way valve.Valveless plunger displacement pump is internal is ceramic material, detectable should not be made to stick in load pump when drawing detectable by it, in order to avoid detecting for a long time and affecting subsequent detection result；And the high accuracy injection measurement pump configuration of three-way valve is more complicated, it can first pass through a valve and detectable be drawn in pump, then detectable is discharged again through another valve, the addition of detectable can accurately be controlled by this kind of load pump, if adopting the high accuracy injection measurement pump of this three-way valve, the detection equipment of the present invention is capable of more quickly and accurately adding of detectable.
Preferably, the structure of the Photoelectric Detection module of the present invention as shown in Figure 4, this Photoelectric Detection module include a light source 111 being arranged on cuvette 200 side and be arranged on cuvette 200 opposite side with light source just to the optical filter 112 arranged and photoelectric sensor 113.Described light source is used for sending detection light beam, described optical filter 112 for through cuvette transmitted through detection light beam filter, described photoelectric sensor 113 for receive filtered filter after detection light beam.Detect the cuvette after light beam irradiating and detecting reagent has fully reacted with detected sample, when detecting, the light source of Photoelectric Detection module can send detection light, detection light irradiates cuvette, material in cuvette absorption portion can detect light, and at cuvette opposite side, detection light is filtered by optical filter, and photoelectric sensor can receive filtered filter after light, then according to sample in cuvette, the absorbance of light determined the content of HCY in sample.
Based on above-mentioned vitro detection equipment, present invention also offers a kind of external detection method by above-mentioned vitro detection device measuring HCY, as it is shown in figure 5, include step:
Step S100, when the first detectable and the detected sample response time in cuvette being detected then, microprocessor controls sample needle moves to cuvette place along mechanical arm, and is directed at cuvette center；
Step S200, microcontroller start-up loading pump, draw the second detectable from the container holding the second detectable, and add it to react in cuvette；
Step S300, determine the content of HCY in detected sample by the absorbance of reacting substance in cuvette.
Before performing step S100, first it is manually adding to be pre-loaded with in the cuvette of the first detectable by detected sample by testing staff, again cuvette is inserted in corresponding Photoelectric Detection module, stir cuvette by Photoelectric Detection module, make detected sample and the first detectable react fully in cuvette.The response time then, what intervalometer can detect arrives, and sends warning, provides prompting.
Wherein, as shown in Figure 6, also include after step S300:
Step S400, microcontroller again control sample needle and move, and are moved to by sample needle in next cuvette, and the second detectable are joined in this cuvette, carry out next group detection.
In order to make the above embodiment of the present invention become apparent from, below in conjunction with HCY detection in concrete detectable to the present invention by colorimetry measurement HCY content a kind of implementation process be specifically described.
The present embodiment adopts homocysteine detection kit to measure for the Quantitative in vitro of human serum, homocysteine in blood plasma HCY.Oxidized form HCY is converted to free HCY, and free HCY reacts generate L-cystathionine with serine under CBS catalysis.L-cystathionine generates again HCY, acetone acid and NH3 under CBL catalysis.The acetone acid that this circular response generates can detect with lactate dehydrogenase L DH and NADH, and NADH is transformed into the speed of NAD and is directly proportional to HCY content in sample.Therefore, the mixed solution that the first detectable R1 is lactic acid dehydrogenase (LD), serine (SER) and NADH that the present embodiment uses, they content in mixed liquor are respectively more than 800KU/L, 1.3mmol/L and 1mmol/L.Second detectable R2 is cystathionine β-synthase (CBS), cystathionine beta-catabolic enzyme (CBL) and stabilizer, and their content in mixed liquor is respectively > 20KU/L, > 10KU/L and appropriate.Detected sample S is sample is Diagnostic Value of Fasting Serum, blood plasma (EDTA, heparin, anticoagulant).When carrying out HCY detection, cuvette has been preinstalled with the first detectable R1 of 150 microlitres, the detected sample S drawing about 8 microlitres from the first container is added dropwise in cuvette, Photoelectric Detection module is mixed, treat that the two reacts 1-5 minute in cuvette, microprocessor controls sample needle moves to the second container holding the second detectable R2 and goes out, therefrom draw about 40 microlitre the second detectable R2, mobile sample needle goes out to cuvette, the the second detectable R2 drawn is joined in cuvette, 37 DEG C hatch 90s after, monitoring 1-3 minute continuously, calculate the change of absorbance per minute, and then determine the content of HCY in detected sample.
In sum, a kind of semi-automatic external adopting colorimetry to measure HCY detection Apparatus and method for provided by the invention, when intervalometer detects the first detectable and the detected sample response time in cuvette then, microprocessor controls sample needle moves to cuvette place, and is directed at cuvette center；Microcontroller start-up loading pump, draws the second detectable from the container holding the second detectable, and adds it to react in cuvette；The content of HCY in detected sample is determined by the absorbance change of reacting substance in cuvette.Present invention achieves the automatic loading of the second detectable, eliminate the artificial operation loading the second detectable, not only save the cost of labor of HCY detection, eliminate manual monitoring link, but also testing result is impacted by the operation error being avoided as much as artificial dropping detectable；The present invention improves the detection efficiency of HCY simultaneously, substantially reduces the detection time of HCY, has wide market prospect.
It should be appreciated that the application of the present invention is not limited to above-mentioned citing, for those of ordinary skills, it is possible to improved according to the above description or convert, all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (4)
1. the semi-automatic external detection equipment that one kind adopts colorimetry to measure HCY, it is characterized in that, including a monitor station, described monitor station is provided with the Photoelectric Detection module of temperature control and agitating function, described Photoelectric Detection module includes an optical filter, described Photoelectric Detection module is placed with the cuvette pre-installing the first detectable, described monitor station is additionally provided with a mechanical arm, load pump and hold the container of the second detectable, described mechanical arm is provided with a sample needle, one end of described load pump connects described sample needle, the other end connects described container, monitor station is additionally provided with the microcontroller for controlling described detection device detection procedure；Described Photoelectric Detection module includes several and is arranged in a linear on monitor station, and described mechanical arm is suitable with the arrangement of Photoelectric Detection module, and sample needle moves along mechanical arm.
2. employing colorimetry according to claim 1 measures the semi-automatic external detection equipment of HCY, it is characterised in that described load pump is dosing pump.
3. employing colorimetry according to claim 2 measures the semi-automatic external detection equipment of HCY, it is characterised in that described dosing pump is plunger displacement pump or syringe pump.
4. employing colorimetry according to claim 1 measures the semi-automatic external detection equipment of HCY, it is characterized in that, described Photoelectric Detection module includes a light source being arranged on cuvette side, is arranged on cuvette opposite side with described light source just to the photoelectric sensor arranged.
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