CN108982383A - lipoprotein cholesterol detection system - Google Patents
lipoprotein cholesterol detection system Download PDFInfo
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- CN108982383A CN108982383A CN201810797381.9A CN201810797381A CN108982383A CN 108982383 A CN108982383 A CN 108982383A CN 201810797381 A CN201810797381 A CN 201810797381A CN 108982383 A CN108982383 A CN 108982383A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
Abstract
The present invention discloses a kind of lipoprotein cholesterol detection system, which includes sample container, the first reagent container, the second reagent container, eluant container, the first reaction module, the second reaction module and detection module；The sample container and the first reagent container is parallel on the inlet pipeline of the first reaction module by pipeline, and the eluant container is connected on the pipeline that sample container is connect with the first reaction module by pipeline；The outlet of first reaction module is connect with the import of the second reaction module by pipeline, and second reagent container is connected on the pipeline that the first reaction module is connected with the second reaction module by pipeline；The outlet end connecting detection module of second reaction module.With higher sensitivity, and anti-interference ability is improved, and is not limited to the conventional sample of detection.
Description
Technical field
The invention belongs to in-vitro diagnosis fields, specifically a kind of for detecting lipoprotein cholesterol map in serum sample
Instrument and reagent system, that is, lipoprotein cholesterol detection system (system for detecting Lipoproteins subfractions).
Background technique
It is well known that the density according to lipoprotein is different, lipoprotein can be divided into chylomicron (CM), extra-low density rouge
Albumen (VLDL), intermediated-density lipoprotein (IDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL).Clinically,
The cholesterol level of all kinds of lipoprotein is usually measured, for instructing diagnosis.Current clinically conventional determining not only has total gallbladder solid
Alcohol (T-CH), there are also LDL-C, HDL-C.Wherein the too high levels of LDL-C are easy to cause atherosclerosis, and it is solid to be commonly called as bad gallbladder
Alcohol, and HDL-C has certain protective effect to blood vessel, has been commonly called as cholesterol.But it has recently been demonstrated that either LDL-C,
Or HDL-C all has heterogeneity, and clinical meaning is also not exactly the same.It is small and dense if LDL-C is segmented into many subfractions
Low density lipoprotein cholesterol (sdLDL-C) is more easier to cause atherosclerosis, also referred to as since its structure is finer and close
For Type B, and then the ability of atharosclerosis is weaker in contrast for big and light low density lipoprotein cholesterol (LLDL-C), again
Referred to as A type.In HDL-C, it can also at least be divided into two kinds of main subfractions, such as big and light HDL2 particle and small and dense
HDL3 particle, some research evidences have shown that the subfraction of measurement HDL can preferably react cardiovascular than only measurement HDL
The risk of disease.
Currently, conventional clinical testing procedure can only all detect one or a few index, can detect simultaneously multiple
Index, or can react lipoprotein cholesterol distribution technology it is very limited, and rest essentially within the scientific research stage, it is really real
It is existing clinical application and few.Wherein electrophoresis is development than a special kind of skill earlier, is often used in the detection of lipoprotein,
It first passes around after lipoprotein is separated by electrophoresis, then is dyed with corresponding method, such as dye lipoprotein, lust of also having illicit sexual relations sterol,
By scanning data collection analysis.This method is since intermediate steps are cumbersome, electrophoretic separation precision and dyeing scanning
Precision problem, so that the preci-sion and accuracy of this method needs to be considered.Another method is nuclear magnetic resonance method, but the method
Technical threshold is higher, not only expensive equipment, and the requirement to operator is high, so this method is more suitable for scientific research purposes,
And promoting in clinical examination has certain difficulty.In contrast, United States Patent (USP) US5284773, US5633168 are disclosed
A kind of (vertical auto profile) detection technique, the technology have certain excellent in terms of precision, accuracy
Gesture.The technology is lipoprotein to be separated using ultracentrifugation, then detect continuous lipoprotein cholesterol with VAP instrument, and then obtain
Lipoprotein cholesterol spectrogram calculates the lipoprotein cholesterol content of each component.Since vertical rotor is utilized in the technology, so that
Centrifugation time greatly shortens, and required sample size is 50ul.But the technology remains certain defect, such as due to
It is to use cholesterol single reagent, so that sample reacts at a touch with reagent, if there is interfering substance also will be direct in sample
Interaction is reacted with this, causes anti-interference ability poor, and then influences testing result, and due to the chromogen in single reagent
The molar absorptivity of substance is lower, causes sensitivity limited.
Summary of the invention
The present invention is directed to the above-mentioned deficiency of the prior art, provides a kind of lipoprotein cholesterol detection system, the detection system
With higher sensitivity, and anti-interference ability is improved, and is not limited to the conventional sample of detection.
In order to solve the above-mentioned technical problem, the technical solution adopted by the present invention are as follows: a kind of lipoprotein cholesterol detection system,
The system includes sample container, the first reagent container, the second reagent container, eluant container, the first reaction module, the second reaction mould
Block and detection module；The sample container and the first reagent container is parallel to the inlet pipeline of the first reaction module by pipeline
On, the eluant container is connected on the pipeline that sample container is connect with the first reaction module by pipeline；Described first
The outlet of reaction module is connect with the import of the second reaction module by pipeline, and second reagent container is connected by pipeline
In on the pipeline that the first reaction module is connected with the second reaction module；The outlet end connecting detection of second reaction module
Module.
Preferably, the first control valve and sensor are provided on the outlet conduit of the sample container, the biography
Sensor is for detecting whether sample has flowed, and first control valve is used to control the folding of outlet conduit, when the sensing
Device detects that complete first control valve of sample flow is closed, when the sensor detects that sample has the first control of then control
Valve processed is opened.
Preferably, the second control valve is provided on the pipeline that the eluant container outlet end connects, when described
The second control valve described in when one control valve is closed is opened so that cleaning solution cleans pipeline.
Preferably, the junction of the sample container and eluant container is provided with the first triple valve, the sample
The junction of container, the first reagent container and the first reaction module is provided with the second triple valve, second reagent container,
The junction of one reaction module and the second reaction module is provided with third triple valve.
Preferably, first pump housing is provided on the outlet conduit of first reagent container, second reagent
Second pump housing is provided on the outlet conduit of container.
Preferably, first reaction module is the pipeline with certain length, and there is function of temperature control；Using this
Structure when liquid flows through the module, can be incubated for the regular hour, carry out reaction, while there is function of temperature control such as to pass through water
Bath, metal bath, gas bath etc. control temperature, and the control of temperature is conducive to reaction and carries out under the conditions of better suited, and can make
Each sample reaction carries out under the same conditions.
Preferably, the duct length of first reaction module is 0.25m-10m, preferably 0.5m-5m, more preferable 1m-
2.5m, most preferably 1.5m.
Preferably, second reaction module is the pipeline with certain length, and there is function of temperature control；Using this
Structure when liquid flows through the module, can be incubated for the regular hour, carry out reaction, while having function of temperature control such as water-bath, gold
Belong to the control temperature such as bath, gas bath, the control of temperature is conducive to reaction and carries out under the conditions of better suited, and can make each sample
This reaction carries out under the same conditions.
Preferably, the duct length of second reaction module is 0.25m-10m, preferably 0.5m-5m, more preferable 1m-
2.5m, most preferably 1.5m.
Preferably, the third pump housing and waste collection tank are connected on the pipeline of the outlet end of the detection module, and
Detection module is also electrically connected with logging modle and storage equipment.Using the above structure, the material come out from detection module is by waste liquid
Collecting tank is timely collected, and not can cause environmental pollution and is destroyed；And logging modle can directly record detection as a result, simultaneously
It is printed, and data can be saved by storing equipment.
Preferably, first reagent container is for storing reagent R1, the reagent R1 main component is surface
Activating agent Lutensol AP 8 (alkylphenol ethoxylate of trading company of man of Haitai Jin nation as above sale, it is therein
Lutensol AP series) and triton x-100,4-AAP (4 amino antipyrine), ascorbic acid oxidase etc.；Described
Two reagent containers are for storing reagent R2；Reagent R2 main component be cholesterol oxidase, cholesterol esterase, peroxidase,
TOOS etc..
Content as further preferred, described reagent R1 main component are as follows: the concentration of Lutensol AP 8 is
0.01%-5% (mass percentage), the concentration of triton x-100 are 0.01%-5% (mass percentage), 4-AAP's
Concentration is 0.1mM-10mM, and the concentration of ascorbic acid oxidase is 0.1KU/L-20KU/L.
Further, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.05%-1%, Qula
The concentration of logical X-100 is 0.05%-1%, and the concentration of 4-AAP is 0.5mM-5mM, and the concentration of ascorbic acid oxidase is 0.5KU/
L-10KU/L。
Further, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.1%-0.5%, Qula
The concentration of logical X-100 is 0.1%-0.5%, and the concentration of 4-AAP is 1mM-2mM, and the concentration of ascorbic acid oxidase is 1KU/L-
5KU/L。
Further, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.1%, and Qula leads to X-
The concentration that 100 concentration is 1.0%, 4-AAP is 1mM, and the concentration of ascorbic acid oxidase is 2KU/L.
As further preferred, the content of the reagent R2 main component are as follows: the concentration of cholesterol oxidase is 0.1-
20KU/L, the concentration of cholesterol esterase are 0.1-20KU/L, and the concentration of peroxidase is 0.1-20KU/L, and the concentration of TOOS is
0.1mM-10mM。
Further, the content of the reagent R2 main component is that the concentration of cholesterol oxidase is 0.5-10KU/L, gallbladder
The concentration of sterol esterase is 0.5-10KU/L, and the concentration of peroxidase is 0.5-10KU/L, and the concentration of TOOS is 0.5mM-
5mM。
Further, the content of the reagent R2 main component is that the concentration of cholesterol oxidase is 1-5KU/L, and gallbladder is solid
The concentration of alcohol esterase is 1-5KU/L, and the concentration of peroxidase is 1-5KU/L, and the concentration of TOOS is 1mM-2mM.
Further, the content of the reagent R2 main component is that the concentration of cholesterol oxidase is 2KU/L, cholesterol
The concentration of esterase is 2KU/L, and the concentration of peroxidase is 2KU/L, and the concentration of TOOS is 2mM.
In addition, in specific preparation of reagents process buffer, preservative etc. can also be added, the reagent of acquisition is more
Stablize, such as buffer can use phosphate buffer (PB), preservative can be using Sodium azide etc..
Main function principle of the invention and the process sample after ultracentrifugation separates as shown in Figure 1:, from Fig. 1
The bottom of sample container enter instrument, while receiving the detection of sensor, once sample drains off, the first control valve will be closed
It closes, enter air will not in the pipe-line system of instrument.Sensor and the first control valve are controlled by controller；When there is sample
When being detected, the first control valve is in the open state, sample can be made to enter pipeline, when a sample sample introduction finishes
Afterwards, the first control valve is closed, and the second control valve is opened, and cleaning solution such as physiological saline enters pipeline in cleaning container, right
Pipeline is cleaned, in case the detection of next sample is prepared.When cleaning finishes, and after next sample is ready,
Second control valve is closed, and the first control valve is opened, and carries out the detection of next sample.
Advantages of the present invention and beneficial aspects:
1. system of the invention, wherein containing reagent R1 in the first reagent container, under the action of first pump housing, with certain
Flow velocity enter in the pipeline of instrument, and mixed at the second triple valve with sample, while the second triple valve can also play sample
Reagent immixture.After reagent is mixed with sample, the first reaction module is entered back into, the first reaction module, which can be one, to be had
The pipeline of certain length when liquid flows through the module, can be incubated for the regular hour, carry out reaction, while having temperature control function
Energy such as water-bath, metal bath, gas bath, the control of temperature are conducive to reaction and carry out under the conditions of better suited, and can make every
A sample reaction carries out under the same conditions.
2. system of the invention, wherein the reagent R2 in the second reagent container is under the action of second pump housing, into instrument
It in pipeline, and is mixed at third triple valve with the liquid come out from the first reaction module, which plays two pipes simultaneously
The effect that reagent mixes in road.Mixed liquid flows to the second reaction module, and the second reaction module, which can be one, has one
The pipeline of measured length when liquid flows through the module, can be incubated for the regular hour, carry out reaction, while having function of temperature control
Such as water-bath, metal bath, gas bath, the control of temperature is conducive to reaction and carries out under the conditions of better suited, and can make each
Sample reaction carries out under the same conditions.
3. system of the invention, wherein after the second reaction module be detection module, usual detection module be can be point
Light photometer, is able to detect the absorbance that the reagent come is flowed through from the second reaction module, and absorbance detected is recorded mould
Block record, and be stored in computer, it is used for further analytical calculation.After the third pump housing is located at detection module, it can will test
Reagent afterwards is rejected in waste collection tank behind.The third pump housing also has the function of the entire fluid path of control simultaneously, with the
One pump housing, second pump housing, the first control valve, the second control valve collective effect, make entire fluid path from sample and reagent in order
Reaction module and detection module are flowed to, can prevent fluid path from flowing backwards.
4. the present invention compares and former instrument system, the second reagent container, second pump housing and the first reaction mould are increased
Block, while original cholesterol single reagent has also been replaced with cholesterol double reagent R1 and R2.R1 main component is surface-active
Agent Lutensol AP 8 and triton x-100,4-AAP, anti-change-blood acid oxidase etc., R2 main component be cholesterol oxidase,
Cholesterol esterase, peroxidase, TOOS etc..This innovative characteristics of the invention sample is mixed with reagent R1 after first
There is certain reaction time in reaction module, is conducive to remove the interfering substance in sample in this way, so that these interfering substances pair
Subsequent reaction influence reaches minimum.When reagent R2 is again and by centainly going the sample of disturbing reaction and reagent R1 mixture to mix
When, the signal of reacted generation is the signal of substance to be detected in sample, not by the interference of other impurity.For example, with
The improvement of people's living standards, it will usually supplement various nutriments, such as vitamins, wherein vitamin C is a kind of
Substance with reproducibility just will appear vitamin C in serum, it can make cholesterol when patient has taken this substance
Reagent reacts generated hydrogen-peroxide reduction with sample and falls, so that the content of hydrogen peroxide is reduced, further, coupling
Coloring matter caused by reacting also is reduced, so that detected value is obviously relatively low.This influence is for testing result
It influences very serious, it should cause enough attention.However system of the invention but can be to avoid this phenomenon, because in R1
It is added to ascorbic acid oxidase, the incubation of the certain time in the first reaction module by R1 and sample, vitamin C can be by
Ascorbic acid oxidase institute oxygenolysis, the product of generation do not have reproducibility, so that entire reaction will not be disturbed, allow detection
As a result more prepare.
5. for the single reagent of the prior art, all necessary raw materials of reaction all can only in a reagent, even if
Cucumber, which mixes, is easy to happen discoloration, degradation etc., also without good solution.Such as showing in cholesterol reagent
Color substance 4-AAP and phenol, both necessary to chromogenic reaction, but for a long time in a solution, oxygen occurs for solution
Change from developing the color, stability is poor.Since present invention uses double reagent systems, unstable substance can be prepared respectively in R1
In reagent or R2 reagent, the stability of reagent can be greatly improved.Simultaneously because the reagent, which can separate, considers unstable object
Phenol is substituted for another chromogen substance TOOS by matter, and the molar absorption coefficient of the substance is formed by coloring matter than phenol
The big many of molar absorption coefficient, therefore the sensitivity of whole system will also greatly improve.System i.e. of the invention is for double reagent
It is first blended in the first reaction chamber with reagent R1 with sample and reacts a period of time, then be blended in the second reaction chamber again with reagent R2
It reaction a period of time, is then detected again by detector.It allows the invention to add certain groups in reagent R1 in this way
After point, there are sufficient time and sample to act in the first reaction chamber, and then removes the interfering substance in sample to the shadow of detection
It rings, such as adds ascorbic acid oxidase, remove interference of the vitamin C to detection in sample, anti-interference ability greatly improved.Together
When, double reagent method is conducive to the sensitivity and stability that improve reagent.Phenol and 4- the amino peace that traditional detection system uses
For than woods, chromogen substance is placed in same reagent, spontaneously may gradually be developed the color, stability is poor, and detects sensitive
It spends limited；And it is of the invention, it is placed in R1 and R2 due to separating chromogen substance, when not detecting, it is not easy to form colored substance
Matter, it is more stable, therefore can use the higher chromogen substance of sensitivity, such as TOOS and 4-AA effectively improve
Detection sensitivity.
Detailed description of the invention
Lipoprotein cholesterol detection system structure principle chart Fig. 1 of the invention.
Fig. 2 is lipoprotein cholesterol map of the invention.
Specific embodiment
The present invention is described in further detail below by embodiment, but the present invention is not limited solely to following embodiment.
It is as shown in Fig. 1: a kind of lipoprotein cholesterol detection system of the invention, the system include the first reagent container 1,
Second reagent container 2, sample container 3, eluant container 4, the first reaction module 5, the second reaction module 6 and detection module 7；It is described
Sample container and the first reagent container be parallel to by pipeline on the inlet pipeline of the first reaction module, the eluant container
It is connected to by pipeline on the pipeline that sample container is connect with the first reaction module；The outlet of first reaction module and the
The import of two reaction modules is connected by pipeline, and second reagent container is connected to the first reaction module and the by pipeline
On the pipeline that two reaction modules are connected；The outlet end connecting detection module of second reaction module.The sample holds
The first control valve 8 and sensor 9 are provided on the outlet conduit of device, the sensor is for detecting whether sample has flowed, institute
The first control valve stated is used to control the folding of outlet conduit, when the sensor detects complete first control of sample flow
Valve processed closes, then controls the unlatching of the first control valve when the sensor detects that sample exists.The eluant container outlet
Hold and be provided with the second control valve 10 on the pipeline of connection, when first control valve is closed described in the second control valve open
So that cleaning solution cleans pipeline.The junction of the sample container and eluant container is provided with the first triple valve
11, the junction of the sample container, the first reagent container and the first reaction module is provided with the second triple valve 12, described
The junction of second reagent container, the first reaction module and the second reaction module is provided with third triple valve 13.Described first
It is provided with first pump housing 14 on the outlet conduit of reagent container, is provided with second on the outlet conduit of second reagent container
The pump housing 15.First reaction module is the pipeline with certain length, and has function of temperature control；Using the structure, liquid
When flowing through the module, the regular hour can be incubated for, make reaction carry out, while have function of temperature control such as pass through water-bath, metal bath,
Gas bath etc. controls temperature, and the control of temperature is conducive to reaction and carries out under the conditions of better suited, and each sample can be made anti-
It should carry out under the same conditions.Second reaction module is the pipeline with certain length, and has function of temperature control；It adopts
With the structure, when liquid flows through the module, it can be incubated for the regular hour, carry out reaction, while there is function of temperature control such as water
Bath, metal bath, gas bath etc. control temperature, and the control of temperature is conducive to reaction and carries out under the conditions of better suited, and can make
Each sample reaction carries out under the same conditions.The third pump housing 16 is connected on the pipeline of the outlet end of the detection module
With waste collection tank 17, and detection module (usual detection module can be spectrophotometer) is also electrically connected with logging modle 18
(record module of commercially available energy usage data record) and storage equipment 19 (such as moveable magnetic disc or computer store equipment).Using
Above structure, the material come out from detection module are timely collected by waste collection tank, not can cause environmental pollution and destroy；
And logging modle can directly record detection as a result, and printed, and data can be saved by storing equipment.
Controller of the present invention can be programmable controller, and sensor setting may be programmed in the duct and with described
Controller Electricity Federation；First control valve and the second control valve be respectively set corresponding position in the duct and with the programmable controller
Electrical connection, sensor are also electrically connected with programmable controller；Wherein, the sensor is used to detect the liquid letter in the pipeline
Breath, and the fluid information is passed into the programmable controller by electric signal；First control valve and the second control
Valve controls the closure or unlatching of corresponding pipeline respectively；The sensor can be flow sensor, detect in liquid line
Fluid information, and fluid information is passed into programmable controller by electric signal；The sensor and control valve by
Controller is controlled, when having sample when being detected, the first control valve be it is in the open state so that sample is entered pipeline, when
After one sample sample introduction, the first control valve is closed, and the second control valve is opened, the physiological saline in physiological saline container
Into pipeline, pipeline is cleaned, in case the detection of next sample is prepared.Above-mentioned controller, sensor and valve are equal
For commercial product, as long as being able to satisfy above-mentioned function of the invention may be incorporated for the present invention.
Reagent R1 is contained in first reagent container of the present invention.Reagent R1 main component of the present invention is table
Face activating agent Lutensol AP 8 and triton x-100,4-AAP, anti-change-blood acid oxidase etc.；Second examination of the present invention
Reagent R2 is contained in agent container, reagent R2 main component is cholesterol oxidase, cholesterol esterase, peroxidase, TOOS
Deng.
Main function principle of the invention and the process sample after ultracentrifugation separates as shown in Figure 1:, in Fig. 1
The bottom of sample container enter instrument, while receiving the detection of sensor, once sample drains off, controller will control first
Control valve will be closed, and enter air will not in the pipe-line system of instrument.Sensor and the first control valve are carried out by controller
Control；When having sample when being detected, the first control valve be it is in the open state, sample can be made to enter pipeline, when one
After sample sample introduction, the first control valve is closed, and the second control valve is opened, and is located at cleaning solution such as physiological saline in cleaning container
Into pipeline, pipeline is cleaned, in case the detection of next sample is prepared.When cleaning finishes, and next sample
After ready, the second control valve is closed, and the first control valve is opened, and carries out the detection of next sample.
Sample in sample of the present invention container is the sample after centrifugal treating, specific operating process and each container
Ingredient, concentration and the operating parameter of interior reagent or medium see below embodiment.
First reaction module and the second reaction module of the invention is a degree of pipeline, and specific duct length is at this
It can be 1.5m with the duct length of the first reaction module in embodiment, the duct length of the second reaction module is 1.5m.
Embodiment 1
Pattern detection result compares
Serum 50ul is taken, KBr (Auto-regulating System of Density of Heavy Medium to the 1.21) dilution of 1950ul is added, mixes.Take a 5.1ml's
Polyallomer quick seal tube is added the physiological saline of 3800ul, then is slowly added into the good serum sample of beforehand dilution from bottom
1200ul.The centrifuge tube is put into Rotor V Ti-65.2 rotor, and is centrifuged with Beckman ultracentrifuge.Parameter: revolving speed=
65000rpm, ω2T=6.6*1010rad2/ sec, temperature=23 DEG C accelerate=6, deceleration=6.
Sample after centrifugation is carefully moved on present system (i.e. sample container), is detected, detection parameters are as follows:
Sample flow 1.0ml/min, reagent R1 flow velocity 0.5ml/min, reagent R2 flow velocity 0.5ml/min.Wherein, reagent used is detected
It is as follows:
As control, take with a serum, pre-treatment is centrifuged in the same way.And sample is carefully moved to former VAP
On instrument (system disclosed in such as US5284773, US5633168), detected, parameter are as follows: sample flow 1.0ml/min,
Reagent R flow velocity 1.0ml/min it is as follows to detect reagent used: 50mM PB, 1mM 4-AAP, 2mM phenol, 2KU/L cholesterol oxygen
Change enzyme, 2KU/L cholesterol esterase, 2KU/L peroxidase, 1.0% triton x-100,0.1%Lutensol AP 8,
0.1% Sodium azide.
Lipoprotein cholesterol map obtained as indicated with 2, and calculates the data for obtaining corresponding component with special-purpose software.
It is shown in Table 1.By map and following testing result as it can be seen that the result and original VAP system of detection are almost without difference.
Table 1
Embodiment 2
The comparison of sensitivity
The serum sample that a LDL value is about 100mg/dL is taken, 6 parts of average mark, carries out sample according to the method for embodiment 1
Processing and centrifugation step.Wherein three parts with former VAP system detection, and in addition three parts with VAP system detection of the present invention.Observe each peak
The size of absorbance at value, and with the mean value calculation detected three times.
Table 2
| Former VAP | VAP of the present invention | Absorbance change | |
| Absorbance at LDL peak value | 0.265 | 0.526 | 198.49% |
| Absorbance at HDL peak value | 0.101 | 0.201 | 199.01% |
| Absorbance at VLDL peak value | 0.021 | 0.041 | 195.24% |
As shown in Table 2, VAP system of the invention, the more former VAP system of detection sensitivity has larger promotion, therefore uses
VAP instrument of the invention is more conducive to the inspection of the low Lipoproteins subfractions of certain comparision contents with higher sensitivity
It surveys.
Embodiment 3
Anti-interference ability compares
A pooled serum is taken, is separately added into the interfering substance of various concentration, and detected by the method in embodiment 1.
Select LDL and HDL detected value therein to be calculated and compared, resulting experimental result be shown in Table 3 (LDL interference--free experiments data) and
Table 4 (HDL interference--free experiments data).
Table 3LDL interference--free experiments data
Table 4HDL interference--free experiments data
It is stronger to can be seen that system rejection to disturbance ability of the invention from above-mentioned two parts of tables.
Claims (10)
1. a kind of lipoprotein cholesterol detection system, the system include sample container, the first reagent container, the second reagent container,
Eluant container, the first reaction module, the second reaction module and detection module；The sample container and the first reagent container passes through
Pipeline is parallel on the inlet pipeline of the first reaction module, and the eluant container is connected to sample container and first by pipeline
On the pipeline of reaction module connection；The outlet of first reaction module is connected with the import of the second reaction module by pipeline
It connects, second reagent container is connected on the pipeline that the first reaction module is connected with the second reaction module by pipeline；
The outlet end connecting detection module of second reaction module.
2. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: the outlet of the sample container
The first control valve and sensor are provided on pipeline, the sensor is for detecting whether sample has flowed, first control
Valve processed is used to control the folding of outlet conduit；Closed when the sensor detects complete first control valve of sample flow,
When the sensor detects that sample has then the first control valve and opens.
3. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: the eluant container outlet end
The second control valve is provided on the pipeline of connection, when first control valve is closed described in the second control valve open so that
Cleaning solution is obtained to clean pipeline.
4. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: the sample container and flushing
The junction of container is provided with the first triple valve, the connection of the sample container, the first reagent container and the first reaction module
Place is provided with the second triple valve, the junction setting of second reagent container, the first reaction module and the second reaction module
There is third triple valve.
5. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: first reagent container
It is provided with first pump housing on outlet conduit, is provided with second pump housing on the outlet conduit of second reagent container.
6. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: first reaction module is
Pipeline with certain length, and there is function of temperature control, the length of pipe of first reaction module is 0.25m-10m, preferably
0.5m-5m, more preferable 1m-2.5m, most preferably 1.5m；Second reaction module is the pipeline with certain length, and is had
There is function of temperature control；The length of pipe of second reaction module be 0.25m-10m, preferably 0.5m-5m, more preferable 1m-2.5m, most
It is preferred that 1.5m.
7. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: the outlet of the detection module
The third pump housing and waste collection tank are connected on the pipeline at end, and detection module is also electrically connected with logging modle and storage equipment.
8. lipoprotein cholesterol detection system according to claim 1, it is characterised in that: first reagent container is used
In holding reagent R1, the main component of the reagent R1 is surfactant Lutensol AP 8 and triton x-100,4-
AAP, anti-change-blood acid oxidase；For holding reagent R2, the main component of the reagent R2 is second reagent container
Cholesterol oxidase, cholesterol esterase, peroxidase, TOOS.
9. lipoprotein cholesterol detection system according to claim 8, it is characterised in that:
The content of the reagent R1 main component are as follows: the concentration of Lutensol AP 8 is 0.01%-5%, triton x-100
Concentration be 0.01%-5%, the concentration of 4-AAP is 0.1mM-10mM, and the concentration of ascorbic acid oxidase is 0.1KU/L-
20KU/L；
Preferably, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.05%-1%, triton x-100
Concentration be 0.05%-1%, the concentration of 4-AAP is 0.5mM-5mM, and the concentration of ascorbic acid oxidase is 0.5KU/L-10KU/
L；
Further, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.1%-0.5%, and Qula leads to X-
100 concentration is 0.1%-0.5%, and the concentration of 4-AAP is 1mM-2mM, and the concentration of ascorbic acid oxidase is 1KU/L-5KU/
L；
Most preferably, the reagent R1 main component is that the concentration of Lutensol AP 8 is 0.1%, triton x-100
The concentration that concentration is 1.0%, 4-AAP is 1mM, and the concentration of ascorbic acid oxidase is 2KU/L.
10. lipoprotein cholesterol detection system according to claim 8, it is characterised in that:
The content of the reagent R2 main component are as follows: the concentration of cholesterol oxidase be 0.1-20KU/L, cholesterol esterase it is dense
Degree is 0.1-20KU/L, and the concentration of peroxidase is 0.1-20KU/L, and the concentration of TOOS is 0.1mM-10mM；
The content of the preferred reagent R2 main component is that the concentration of cholesterol oxidase is 0.5-10KU/L, cholesteryl ester
The concentration of enzyme is 0.5-10KU/L, and the concentration of peroxidase is 0.5-10KU/L, and the concentration of TOOS is 0.5mM-5mM；
Further, the content of the reagent R2 main component is that the concentration of cholesterol oxidase is 1-5KU/L, cholesteryl ester
The concentration of enzyme is 1-5KU/L, and the concentration of peroxidase is 1-5KU/L, and the concentration of TOOS is 1mM-2mM；
Most preferably, the content of the reagent R2 main component is that the concentration of cholesterol oxidase is 2KU/L, cholesterol esterase
Concentration be 2KU/L, the concentration of peroxidase is 2KU/L, and the concentration of TOOS is 2mM.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810797381.9A CN108982383A (en) | 2018-07-19 | 2018-07-19 | lipoprotein cholesterol detection system |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810797381.9A CN108982383A (en) | 2018-07-19 | 2018-07-19 | lipoprotein cholesterol detection system |
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| CN108982383A true CN108982383A (en) | 2018-12-11 |
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|---|---|---|---|
| CN201810797381.9A Pending CN108982383A (en) | 2018-07-19 | 2018-07-19 | lipoprotein cholesterol detection system |
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