CN114965653B - Double-cabin photoelectrochemical sensing system for detecting prothrombin time and detection method - Google Patents
Double-cabin photoelectrochemical sensing system for detecting prothrombin time and detection method Download PDFInfo
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- 238000001514 detection method Methods 0.000 title claims abstract description 78
- 102100027378 Prothrombin Human genes 0.000 title claims abstract description 37
- 108010094028 Prothrombin Proteins 0.000 title claims abstract description 37
- 229940039716 prothrombin Drugs 0.000 title claims abstract description 37
- 238000006243 chemical reaction Methods 0.000 claims abstract description 38
- 239000000463 material Substances 0.000 claims abstract description 7
- 238000000835 electrochemical detection Methods 0.000 claims abstract description 6
- 230000015271 coagulation Effects 0.000 claims description 15
- 238000005345 coagulation Methods 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 11
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- 238000006557 surface reaction Methods 0.000 claims 1
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 abstract description 12
- 239000010931 gold Substances 0.000 abstract description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 abstract description 7
- 229910052737 gold Inorganic materials 0.000 abstract description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract description 6
- 229910052697 platinum Inorganic materials 0.000 abstract description 6
- 229910052799 carbon Inorganic materials 0.000 abstract description 5
- ZOMNIUBKTOKEHS-UHFFFAOYSA-L dimercury dichloride Chemical class Cl[Hg][Hg]Cl ZOMNIUBKTOKEHS-UHFFFAOYSA-L 0.000 abstract description 5
- 229910052724 xenon Inorganic materials 0.000 abstract description 4
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 abstract description 4
- 229910021607 Silver chloride Inorganic materials 0.000 abstract description 3
- 229910021397 glassy carbon Inorganic materials 0.000 abstract description 3
- AMGQUBHHOARCQH-UHFFFAOYSA-N indium;oxotin Chemical compound [In].[Sn]=O AMGQUBHHOARCQH-UHFFFAOYSA-N 0.000 abstract description 3
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 abstract description 3
- 229910052753 mercury Inorganic materials 0.000 abstract description 3
- 239000010453 quartz Substances 0.000 abstract description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 abstract description 3
- 229910052709 silver Inorganic materials 0.000 abstract description 3
- 239000004332 silver Substances 0.000 abstract description 3
- HKZLPVFGJNLROG-UHFFFAOYSA-M silver monochloride Chemical compound [Cl-].[Ag+] HKZLPVFGJNLROG-UHFFFAOYSA-M 0.000 abstract description 3
- 230000004044 response Effects 0.000 description 9
- 229910010413 TiO 2 Inorganic materials 0.000 description 7
- 230000023555 blood coagulation Effects 0.000 description 6
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 5
- 239000001509 sodium citrate Substances 0.000 description 5
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- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 206010053567 Coagulopathies Diseases 0.000 description 1
- 108010090444 Innovin Proteins 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
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- 239000002105 nanoparticle Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000013186 photoplethysmography Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- G01N27/416—Systems
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
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Abstract
The invention relates to the technical field of medical equipment, in particular to a double-cabin photoelectrochemical sensing system for detecting prothrombin time and a detection method. The double-cabin photoelectrochemical sensing system comprises a light source system, a reaction cavity, a detection cavity, a small portable electrochemical detection system, a working electrode, a reference electrode and a counter electrode. The light source system can be an ultraviolet lamp light source such as a large-volume xenon lamp, a mercury lamp and the like, and also can be a miniaturized LED light source, and the power of the light source is adjustable; the reaction cavity and the detection cavity are made of quartz materials, the reaction cavity and the detection cavity can be cubic or cylindrical in shape, and the light source, the reaction cavity and the detection cavity are arranged in a straight line; the working electrode can be a transparent Indium Tin Oxide (ITO) electrode, a glassy carbon electrode, a gold electrode and a platinum electrode; the counter electrode can be a carbon electrode, a gold electrode or a platinum electrode; the reference electrode may be a saturated calomel electrode, a silver/silver chloride electrode. In addition, the invention also provides a prothrombin time detection method based on the double-cabin photoelectrochemical sensing system.
Description
Technical Field
The invention relates to the technical field of medical equipment, in particular to a double-cabin photoelectrochemical sensing system for detecting prothrombin time and a detection method.
Background
Prothrombin Time (PT) is an important index for clinically evaluating blood coagulation function, and has high medical application value in diagnosis and treatment of blood coagulation dysfunction diseases and real-Time evaluation of blood coagulation function in the operation process. At present, the prothrombin time examination in hospitals is mainly performed on a large-scale full-automatic or semi-automatic coagulation analyzer, the reliability of the large-scale instrument is high, the flux is high, the result repeatability is good, but the instrument equipment is expensive, the detection cost is high, and the method is difficult to widely popularize in a basic medical structure with a small sample size. The miniaturized instant detection (Point of Care Testing, POCT) device for blood coagulation has the advantages of low cost and simple operation, and is suitable for conditional basic medical institutions, but the instant detection device for blood coagulation needs to be further improved in the aspects of instrument and device volume, cost, accuracy and the like, so that new detection methods and detection systems are required to be researched, and the instant detection device for blood coagulation with more excellent development performance is required.
Disclosure of Invention
In order to overcome the defects, the invention provides a double-cabin photoelectrochemical sensing system for detecting prothrombin time and a detection method.
The technical scheme for achieving the aim of the invention is as follows:
a dual-cabin photoelectrochemical sensing system for detecting prothrombin time comprises a light source system, a reaction cavity, a detection cavity, a small portable electrochemical detection system, a working electrode, a reference electrode and a counter electrode.
The light source system is an ultraviolet lamp light source such as a large-volume xenon lamp, a mercury lamp and the like, and can also be a miniaturized LED light source, and the power of the light source is adjustable.
Moreover, the reaction cavity and the detection cavity are made of quartz materials, the reaction cavity and the detection cavity can be in a cubic or cylindrical shape, and the light source, the reaction cavity and the detection cavity are arranged in a straight line.
The working electrode is transparent indium tin oxide electrode, glassy carbon electrode, gold electrode and platinum electrode, and the counter electrode can be carbon electrode, gold electrode and platinum electrode. The reference electrode may be a saturated calomel electrode, a silver/silver chloride electrode.
A prothrombin time detection method based on a double-cabin photoelectrochemical sensing system comprises the following steps:
a. adding a proper amount of reaction liquid into the detection cavity, and placing a working electrode modified with a photoelectrochemical sensitive material, a reference electrode and a counter electrode into the reaction liquid of the detection cavity;
b. adding a proper amount of plasma to be detected into the reaction cavity, opening an electrochemical detection system, applying a constant potential on the surface of the working electrode, and recording the current on the surface of the working electrode;
c. turning on a light source to ensure that light is transmitted to the surface of the working electrode through plasma to be detected in the reaction cavity, and continuously recording the current on the surface of the working electrode;
d. when the current on the surface of the working electrode reaches a constant value, adding a proper amount of prothrombin time detection reagent into the reaction cavity, reacting for a period of time, and continuously recording the current on the surface of the working electrode;
e. and reading a time signal of a corresponding reaction coagulation process according to the change trend of the reaction current signal on the surface of the working electrode along with time to obtain a prothrombin time detection result.
A signal processing algorithm characterized by: let Δt=t 4 -T 3 After data processing is carried out on Tmax-s and DeltaT, a new prothrombin time parameter T is obtained n ，T n =tmax-s+b Δt, wherein b ranges from 0.1 to 0.5, the value of b being optimized according to the characteristics of the whole photoelectrochemical sensing system; establish y (PT) =f (T n ) The function relation between the two to obtain the detection data of prothrombin time; the prothrombin time detection reagent is added at the moment T3 to start the coagulation reaction, the plasma coagulation scatters the light, the light intensity transmitted to the surface of the working electrode is reduced, the photocurrent begins to be obviously reduced, and the plasma coagulation at the moment T4The basic reaction is completed, the photocurrent is slowly reduced, the time point Tmax-s corresponding to the maximum slope point of the photocurrent-time change curve between the time point T3 and the time point T4 is taken,
the invention has the advantages that:
1. the invention provides a signal processing algorithm for improving the precision and accuracy of detecting prothrombin time by an electrochemical sensing system, and establishes y (PT) =f (T n ) And the function relation between the two is used for obtaining the detection data of prothrombin time, so that the precision and the accuracy of a detection system are improved.
2. The invention improves the detection performance of the photoelectrochemical sensing system through algorithm optimization. Compared with an algorithm adopting Tmax-s as a detection signal, the Tn is adopted as the detection signal, so that the accuracy and precision of detection can be obviously improved, and the influence caused by the difference of working electrodes can be reduced
Drawings
FIG. 1 is a schematic diagram of a dual-compartment photoelectrochemical sensing system for detecting prothrombin time;
FIG. 2 is an exemplary plot of photoplethysmography versus reaction time for prothrombin time;
FIG. 3 is a calibration curve obtained by fitting the PT values detected by SYSMEX CA-1500 with Tmax-s as the detection signal in the example.
FIG. 4 is a calibration curve obtained by fitting the detection signal Tn to the PT value detected by SYSMEX CA-1500.
The device comprises a 1-light source system, a 2-reaction cavity, a 3-detection cavity, a 4-small portable electrochemical detection system, a 5-working electrode, a 6-reference electrode and a 7-counter electrode.
Detailed Description
In one aspect, the present invention provides a dual-cabin photoelectrochemical sensing system for detecting prothrombin time, a schematic diagram of the dual-cabin photoelectrochemical sensing system is shown in fig. 1, the light source system may be an ultraviolet light source such as a large-volume xenon lamp, a mercury lamp, or a miniaturized LED light source, and the power of the light source is adjustable.
The reaction cavity and the detection cavity are made of quartz materials, the reaction cavity and the detection cavity can be cubic or cylindrical in shape, and the light sources, the reaction cavity and the detection cavity are arranged in a straight line. The working electrode may be a transparent Indium Tin Oxide (ITO) electrode, a glassy carbon electrode, a gold electrode, or a platinum electrode. The counter electrode can be a carbon electrode, a gold electrode and a platinum electrode, and the reference electrode can be a saturated calomel electrode and a silver/silver chloride electrode.
The invention provides a detection method for detecting prothrombin time by adopting the double-cabin photoelectrochemical sensing system, which has the following basic principle: the surface of the working electrode is decorated with photoelectrochemical sensitive materials, when a certain voltage is applied to the surface of the working electrode, when light with a certain wavelength is irradiated to the surface of the working electrode, the response photocurrent of the surface of the working electrode is related to the energy of the light irradiated to the surface of the electrode. When the light source is not turned on, light irradiates the working electrode surface in the detection cavity through the reaction cavity, photocurrent response is obviously increased, prothrombin time detection reagent is added after the photocurrent response is stabilized, coagulation reaction is started, the plasma to be detected is gradually coagulated, partial light is scattered by the coagulated plasma, so that light energy transmitted to the working electrode surface in the detection cavity is gradually reduced, response photocurrent of the working electrode surface is reduced, and the relationship of photocurrent response changing with time can reflect coagulation process.
The detection method comprises the following detection steps: step 1: adding a proper amount of reaction liquid into the detection cavity, and placing a working electrode modified with a photoelectrochemical sensitive material, a reference electrode and a counter electrode into the reaction liquid of the detection cavity; step 2; adding a proper amount of plasma to be detected into the reaction cavity, opening an electrochemical detection system, applying a constant potential on the surface of the working electrode, and recording the current on the surface of the working electrode; step 3: turning on a light source to ensure that light is transmitted to the surface of the working electrode through plasma to be detected in the reaction cavity, and continuously recording the current on the surface of the working electrode; step 4: when the current on the surface of the working electrode reaches a constant value, adding a proper amount of prothrombin time detection reagent into the reaction cavity, reacting for a period of time, and continuously recording the current on the surface of the working electrode; step 5: and reading a time signal of a corresponding reaction coagulation process according to the change trend of the reaction current signal on the surface of the working electrode along with time to obtain a prothrombin time detection result.
Fig. 2 is an exemplary plot of photo-current versus reaction time change for a sensing system of the present invention detecting prothrombin time in accordance with an embodiment. When the light source is turned on in the time T1, the photocurrent is obviously increased, the time T2 reaches a stable state, the prothrombin time detection reagent is added in the time T3, the coagulation reaction is started, the plasma coagulation causes the light to scatter, the light intensity transmitted to the surface of the working electrode is reduced, the photocurrent starts to be obviously reduced, the basic reaction of the plasma coagulation reaction is completed in the time T4, and the photocurrent is slowly reduced. And (3) taking a time point (Tmax-s) corresponding to the maximum slope point of the photocurrent-time change curve between the time point T3 and the time point T4, and establishing a functional relation between y (PT) =f (Tmax-s) to obtain detection data of prothrombin time.
In order to further improve the precision and accuracy of the photoelectrochemical sensing system for detecting prothrombin time, the invention also provides a signal processing algorithm, which comprises the following specific contents: let Δt=t 4 -T 3 After data processing is carried out on Tmax-s and DeltaT, a new prothrombin time parameter T is obtained n ，T n =tmax-s+b Δt, wherein b has a value ranging from 0.1 to 0.5, and b can be optimized according to the characteristics of the entire photoelectrochemical sensing system. Establish y (PT) =f (T n ) And the function relation between the two is used for obtaining the detection data of prothrombin time, so that the precision and the accuracy of a detection system are improved.
Example 1:
the sensor system is used for detecting the PT value of the clinical sample.
1. Preparation of working electrode for photoelectrochemical detection
The process is as follows: (1) Taking commercial TiO 2 3mg of nano particles (P25, desolid race) are dispersed in 2ml of water by ultrasonic, 0.1mg of graphene (GR, xianfeng nanometer) is added into the water, and the mixture is subjected to ultrasonic treatment for 30 minutes to obtain TiO 2 -a mixed solution of GR dispersed homogeneously; (2) Taking 100 mu LTiO 2 GR dispersion is dripped on the surface of the pretreated clean ITO electrode, and after the ITO electrode is dried at room temperature, the ITO electrode is protected by argonHeating at 450deg.C for 60min, naturally cooling to room temperature to obtain TiO 2 -GR/ITO modified electrode; (3) TiO is mixed with 2 The GR/ITO modified electrode is placed in 0.5mol/LHAuCl4+0.1mol/L PBS (pH 7.0) solution, saturated calomel is used as reference electrode, carbon rod is used as counter electrode, and the solution is prepared by using a catalyst in the presence of TiO 2 Applying potential-0.4V on the surface of the GR/ITO modified electrode, depositing for 100s, and depositing nano gold particles on the surface of the electrode to obtain Au/TiO 2 GR/ITO modified electrode.
2. Detection cavity reaction system assembly
Au/TiO 2 GR/ITO electrode, saturated calomel electrode, carbon rod electrode were placed in the detection chamber, and 2mL of 0.1mol ascorbic acid+0.1 mol/L PBS (pH 7.4) solution was added to the detection chamber, ensuring that all electrodes were immersed in the solution.
3. And selecting proper sodium citrate anticoagulated plasma from a hospital, and carrying out PT detection on a clinical sample by adopting the sensing system.
The process is as follows: (1) Adding 0.2mL sodium citrate anticoagulated plasma into the reaction cavity, adding the sodium citrate anticoagulated plasma into Au/TiO 2 -applying a potential of 0V to the GR/ITO electrode surface; (2) Turning on a light source (xenon lamp), adding 0.4mL commercial PT detection reagent (Dade Innovin reagent) into the reaction cavity after the current response is stable, and recording the change relation of photocurrent along with time; (3) Measuring photocurrent responses of sodium citrate anticoagulated plasma samples under different response times to obtain time parameters T3, tmax-s and T4 of the photocurrent responses, and adopting a formula T n Tmax-s+b Δt, b=0.25, and Tn is calculated. In addition, all sodium citrate anticoagulated plasma samples were tested for PT values using the SYSMEX CA-1500 clotting assay system. The results obtained are shown in Table 1:
TABLE 1 detection results
As can be seen from the data in Table 1, the fitting of the detection signal Tmax-s to the PT value detected by SYSMEX CA-1500 was performed in the range of 9.1-98.2s (FIG. 3), and the correction curve obtained was y (PT) = 1.9273x (DeltaT) -10.078, R 2 = 0.9849 with Tn as the detection signal and SYThe PT values detected by SMEX CA-1500 were fitted (FIG. 4), and the resulting correction curve was y (PT) = 1.3172x (Tn) -5.3027, R 2 The correlation is good in the range of=0.9914, which indicates that the sensing system and the detection method provided by the invention can be used for detecting prothrombin time of clinical samples.
Example 2:
when a plurality of working electrodes for photoelectrochemical sensing are prepared by the method in example 1, the performance of the working electrodes may be different due to the instability of the preparation process flow, and the detection performance of the whole photoelectrochemical sensing system is affected, resulting in the reduction of the detection accuracy and precision of the system. In order to improve the stability of the photoelectrochemical sensing system and improve the accuracy and precision of detection, the invention improves the detection performance of the photoelectrochemical sensing system through algorithm optimization. Compared with an algorithm using Tmax-s as a detection signal, the method using Tn as the detection signal can remarkably improve the accuracy and precision of detection, reduce the influence caused by the difference of working electrodes, and the related results are shown in Table 2.
TABLE 2
The above embodiments are only for the preferred embodiments of the present invention and are intended to illustrate the principle and the function of the present invention, not to limit the present invention. Modifications, substitutions, or improvements of the present invention may be made by those skilled in the art without departing from the spirit and principles of the present invention. Accordingly, the scope of protection of the present patent is intended to cover the appended claims.
Claims (1)
1. A prothrombin time detection method based on a double-cabin photoelectrochemical sensing system is characterized by comprising the following steps of: the method comprises the following steps:
a. adding a proper amount of reaction liquid into the detection cavity, and placing a working electrode modified with a photoelectrochemical sensitive material, a reference electrode and a counter electrode into the reaction liquid of the detection cavity;
b. adding a proper amount of plasma to be detected into the reaction cavity, opening an electrochemical detection system, applying a constant potential on the surface of the working electrode, and recording the current on the surface of the working electrode;
c. turning on a light source to ensure that light is transmitted to the surface of the working electrode through plasma to be detected in the reaction cavity, and continuously recording the current on the surface of the working electrode;
d. when the current on the surface of the working electrode reaches a constant value, adding a proper amount of prothrombin time detection reagent into the reaction cavity, reacting for a period of time, and continuously recording the current on the surface of the working electrode;
e. according to the change trend of the working electrode surface reaction current signal along with time, reading a time signal of a corresponding reaction coagulation process to obtain a prothrombin time detection result;
the signal processing algorithm for the detection method is that delta T=T 4 -T 3 After data processing is carried out on Tmax-s and delta T, a new prothrombin time parameter T is obtained n ，T n =tmax-s+b Δt, wherein b has a value ranging from 0.1 to 0.5, the value of b being optimized according to the characteristics of the whole photoelectrochemical sensing system; establish y (PT) =f (T n ) The function relation between the two to obtain the detection data of prothrombin time; and adding a prothrombin time detection reagent at the moment T3, starting a coagulation reaction, enabling plasma coagulation to scatter light, reducing the light intensity transmitted to the surface of the working electrode, and enabling the photocurrent to be obviously reduced, wherein at the moment T4, the plasma coagulation reaction is basically finished, the photocurrent is slowly reduced, and taking a time point Tmax-s corresponding to the maximum slope point of a photocurrent-time change curve between the moment T3 and the moment T4.
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| CN109507260A (en) * | 2018-12-29 | 2019-03-22 | 中国科学院苏州生物医学工程技术研究所 | Electrochemical Detection chip, electrochemical sensor and its preparation method and application |
| CN112748171A (en) * | 2020-03-24 | 2021-05-04 | 湖南工业大学 | Blood coagulation function index detection test paper, preparation and signal processing method thereof |
| CN112098485A (en) * | 2020-09-16 | 2020-12-18 | 山东科技大学 | Photoelectrochemical aptamer sensor based on sensing separation strategy and preparation method and application thereof |
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