CN214895325U - Homogeneous immunoassay's instant detecting system - Google Patents

Abstract

The utility model discloses a homogeneous immunoassay's instant detecting system, this system include light source, photoelectric control system, mixing system, temperature control system, reaction cup, detecting system, data collection processing system. The system adopts a double-light-path detection design, can compatibly use a latex enhanced turbidimetric immunoassay reagent and a homogeneous luminescent immunoassay reagent for detection, and solves the problems of compatibility, universality and equipment integration of an instant detection platform aiming at the latex reagent and the homogeneous luminescent reagent in the technical field of immunodiagnosis. Meanwhile, the system of the invention has compact design, does not need liquid path design, has no mechanical moving part, reduces the cost of the instrument, reduces the volume of the instrument, improves the stability and the durability of the instrument, has simple and convenient operation and quick detection, and meets the requirement of the POCT instant detection system.

Description

Homogeneous immunoassay's instant detecting system

Technical Field

The invention relates to the technical field of immunodiagnosis, in particular to a real-time detection system for homogeneous immunodiagnosis analysis.

Background

Immunodiagnosis is a diagnostic method that is currently frequently used clinically. Common immunodiagnostic assays can be classified into enzyme-linked immunosorbent assay (ELISA), fluorescence immunochromatography, latex-enhanced immunoturbidimetry, chemiluminescence immunoassay, and the like according to the technical principles. These analysis techniques can be divided into two major categories, i.e., homogeneous immunoassay and heterogeneous immunoassay, depending on whether the reaction system needs to be separated or washed, wherein latex-enhanced immunoturbidimetry and homogeneous chemiluminescence immunoassay are the most common homogeneous immunoassay techniques at present.

The latex enhanced immunoturbidimetric reagent is a homogeneous immunological reagent adopting a latex enhanced immunoturbidimetric Assay (Particles enhanced immunoturbidimetric immunoassay-Assay-PETIA). Latex enhanced immunoturbidimetry (PETIA) specifically captures an analyte in a test sample by immunoreaction using an antibody or antigen adsorbed on latex microparticles having a diameter of several tens to several hundreds nanometers, and forms cross-linking between microparticles and aggregation of microparticles by bridging of the captured analyte. The agglomeration of the particles increases the scattering or transmission absorbance (i.e., turbidity) of the reagent. The concentration of the substance to be detected in the test sample is quantitatively calculated from the calibration curve by measuring the change of turbidity with time. The detection method does not need a long-time incubation process and a washing and separating step, and the whole detection process is finished in a homogeneous system, so the detection speed is high, the operation is relatively simple and convenient, and the method is widely applied to clinic. The test method can be applied to a general large-scale automatic biochemical analyzer and can also be applied to small-scale POCT (point of care testing) equipment such as a specific protein analyzer and the like, has strong applicability, and can well meet the use requirements from a large-scale three-hospitals to community primary hospitals in China.

The homogeneous phase chemiluminescence immunoassay reagent is a novel chemiluminescence detection reagent developed according to LOCI (luminescence oxygen channel assay) technology invented in 1994. This reagent contains two types of microspheres. One is an oxygen-donating microsphere containing a singlet oxygen generating compound such as a phthalocyanine molecule; one is an oxygen-accepting microsphere, which contains a fluorescent substance capable of accepting singlet oxygen. The oxygen supply microsphere can generate a large amount of singlet oxygen molecules under the excitation of light with the wavelength of about 680 nm. The half-life of singlet oxygen is only 4us, the singlet oxygen is quickly attenuated in an aqueous medium, and the singlet oxygen can be transmitted at a distance of about 200nm farthest. If the oxygen-receiving microsphere can approach the oxygen-supplying microsphere through specific recognition such as immune recognition, the fluorescent group in the oxygen-receiving microsphere can rapidly absorb singlet oxygen generated under 680nm light excitation, and simultaneously generate 520-620nm optical signals. Therefore, when the oxygen supplying microsphere and the oxygen receiving microsphere are coated with the proper paired antibodies, antigens or other specific molecules and can form bridging and condensation through the corresponding analyte in the test sample, under the excitation of 680nm light, 520-620nm light signals can be generated, and the intensity of the signals and the amount of the analyte in the test sample are in a linear relationship in a certain range. By detecting the intensity of the emitted light and then comparing the calibration curve, the concentration of the analyte (here, the antigen) in the sample can be quantitatively detected. Similar to the latex-enhanced turbidimetry method mentioned above, the detection process of this chemiluminescence immunoassay method is also performed in a homogeneous system, and does not require washing separation, and thus is called a homogeneous chemiluminescence immunoassay method.

The homogeneous immunoassay technique does not need a separation reaction system, and avoids a cleaning process, so the method has high detection speed generally and is particularly suitable for a point-of-care testing system (POCT). However, the existing latex enhanced immunoturbidimetric assay reagent can only be used in an instant detection platform of a latex reagent, while a homogeneous chemiluminescent reagent can only be used in an instant detection platform of a homogeneous luminescent reagent, and the reagent has no universality among platforms, and the platforms have no compatibility with the reagent. However, in actual use, due to the reasons of reagent cost, detection sensitivity, linear range of detection and the like, the detection items of the two types of reagents are often different from each other, and it is impossible to detect all items on one platform. In order to cover more detection items, two instant detection platforms need to be equipped at the same time, which causes resource waste and also makes the convenience of using a homogeneous reagent for instant detection discount.

Disclosure of Invention

In order to solve the problems of compatibility, universality and integration of a latex reagent and a homogeneous luminescent reagent of an instant detection platform, the invention designs a universal instant detection system of homogeneous immunoassay, which can simultaneously use a latex enhanced turbidimetric immunoassay reagent and a homogeneous chemiluminescent immunoassay reagent.

The invention provides a real-time detection system of homogeneous immunoassay universal for a latex enhanced immunoturbidimetric reagent and a homogeneous chemiluminescence immunoassay reagent. The system comprises: the device comprises a light source, a photoelectric control system, a stirring system, a temperature control system, a reaction cup, a detection system module, a data collection and processing system and a power supply system;

optionally, the light source may be a laser light source, or may be a halogen lamp light source, a laser diode, or an LED light source;

optionally, the optoelectronic control system may be used to control optoelectronic signals within the system;

optionally, the stirring system may be a magnetic stirring system, and the magnet or the electromagnet is driven by a motor to move, so that the magnetic stirrer in the reaction cup moves to stir and mix the solution in the reaction cup;

optionally, the temperature control system can be used for controlling the temperature of the whole detection system including the reaction cup system;

optionally, the light-transmitting part of the reaction cup is made of a transparent material, and may be a polymer material or an inorganic material such as glass or quartz;

optionally, the reaction cup may be pre-filled with a reaction reagent, or a reaction reagent may be added during the detection process;

optionally, the detection system module comprises two or more light sensors; the two light sensors may be the same or different;

optionally, the light sensor may be disposed right in front of the incident light path, or may be disposed at an angle with respect to the incident light path. This angle may be an acute angle, a right angle, or an obtuse angle. Optionally, a lens or an optical filter may be disposed in front of the optical sensor;

optionally, the data collection and processing system may be connected to the detection module, collect the optical signal and convert the optical signal into a digital signal, and perform subsequent processing by pre-installed software;

alternatively, the power system may be used to provide power for the instant detection system, and may be a battery, or other power source.

The homogeneous immunoassay real-time detection system designed by the invention has the following advantages:

1. the universal homogeneous immunoassay instrument can be compatible with a latex enhanced immunoturbidimetric reagent and a homogeneous chemiluminescent immunoassay reagent;

2. the design is compact, a liquid path is not needed, no mechanical moving part is arranged, the cost of the instrument is reduced, the volume of the instrument is reduced, and the stability and the durability of the instrument are improved;

3. the reaction cup can be used for pre-loading reagents, so that the material consumption is reduced, and the inspection process is simplified;

4. the method is simple and convenient to operate, is quick to detect, can finish detection within 3 to 5 minutes and display results, and accords with the concept of quick and simple instant detection.

Drawings

The following describes a specific embodiment of the present invention with reference to the drawings. It is to be expressly understood that the drawings described below are illustrative of some specific embodiments of the invention and are not intended as the only embodiments of the invention. Other forms of embodiment can be derived by those skilled in the art from these figures without inventive effort.

Fig. 1 is a perspective view of an instant detection system according to an embodiment of the present invention.

Fig. 2 is a schematic diagram of an internal structure of an instant detection system according to an embodiment of the present invention.

Fig. 3 is a schematic diagram of the dual optical path detection of the instant detection system according to the embodiment of the present invention.

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention will be described in further detail with reference to specific examples. The following examples are intended to further illustrate preferred embodiments of the invention, and are not intended to be exhaustive. Other embodiments of the invention based on the present invention, which can be made by those skilled in the art without inventive efforts, fall within the scope of the present invention.

Example 1

The invention relates to a real-time detection system compatible with a latex enhanced immunoturbidimetric reagent and a homogeneous chemiluminescence immunoassay reagent. From the schematic of fig. 1 and 2, it can be seen that the whole device is very compact, with dimensions of 15cm × 10cm × 7cm (length × width × height), and a compact design. Specifically, in this embodiment, the light source a may be a halogen lamp light source, an LED light source, or a laser light source such as a laser diode. Preferably, the light source is a laser diode that generates laser light having a wavelength of 680 nm. During detection, light generated by the light source irradiates a reaction system in the reaction cup.

According to the illustration of fig. 2, the light sensor of the optical path front detection module b1 is used to receive transmitted or scattered light. The light sensor positioned directly in front of the light path makes an angle of less than 3 degrees with the incident light and can be used to receive the transmitted light. The light sensor may also be offset from the light path at an angle of greater than 3 degrees from the incident light to receive scattered light. Preferably, the light sensor is angled at 5 degrees to the incident light. The lens is additionally arranged in front of the sensor, so that light signals can be collected more effectively. The lens is provided with a filter in front of the lens, and preferably, the filter can be provided with a filter of 500 nm and 700 nm.

According to the illustration of fig. 2, the light sensor of the optical path vertical direction detection module b2 is used to receive the chemiluminescence signal. The lens is additionally arranged in front of the sensor, so that light signals can be collected more effectively. The detection module b2 also contains a photomultiplier tube for amplifying the received optical signal. The lens for collecting the optical signal is provided with a filter in front, and preferably, a filter for passing 520-620nm wavelength can be arranged.

Referring to fig. 2, the internal structure of the system further comprises a reaction cup c, a stirring system d and a temperature control system e. The reaction cup groove is used for placing a reaction cup, and the size can be set according to the reaction cup. Preferably, a square slot of 1cm by 1cm may be provided. The stirring system d comprises a control system, a motor, a connecting rod and a magnet. The control system controls the operation of the motor. The motor drives the connecting rod to move, and the connecting rod drives the magnet to move up and down. The up-and-down movement of the magnet can drive the stirrer in the reaction cup to move up and down, thereby stirring and mixing the solution in the reaction cup. The temperature control system e comprises a control system and an electric heating plate. The control system can detect the ambient temperature of the system and start or stop the heating of the electric heating plate.

Referring to fig. 2, the instant detection system further comprises a reaction cup. The reaction cup can be a disposable material as a consumable material of the system. The cuvette is required to be made of a transparent material at least at the position of the optical path. It can be various common transparent high molecular materials, various silicon-based inorganic materials such as silicate, etc., and natural transparent materials such as crystal, etc. Preferably, a polypropylene material may be used. The volume of the reaction cup can be variously selected from 300uL, 500uL, 1mL, 2mL and the like, and preferably, the volume of the reaction cup is 1 mL. The reaction cup can be pre-filled with reaction reagents, and the reagents can be filled according to the requirements of the reagents during detection. A stirrer is arranged in the reaction cup. The stirring bar may be of various geometric shapes of material that can be attracted by a magnet. Preferably 304 stainless steel balls. The stainless steel balls can be coated with inert anticorrosive coatings.

The system also comprises a photoelectric control system f which comprehensively controls the light source, the detection module, the stirring system, the temperature control system and the data collection and processing system under the control of preset software.

The data collecting and processing system of the system is used for collecting photoelectric signals collected by the detection module, making mathematical conversion according to the setting of preset software, submitting data to human-computer interface software or sending the data to other peripheral equipment through an output port.

Referring to fig. 1, the system also has control software and a display screen g installed in advance. The display screen is a touch screen. The control software operates on the display screen, and the detection result is also displayed on the display screen.

Referring to fig. 1, the system may also be externally connected to a printing device. Preferably bluetooth or other wirelessly connected printing device.

The detection process of the present embodiment is briefly described below. And starting a power supply, and enabling the system to enter a self-checking mode to ensure that each subsystem works normally. And after the self-checking is finished, the temperature control module is started, and the temperature control target is 37 ℃. When the detection is started, a corresponding detection item is selected on the display operation interface. The reaction cup is filled with reagents, a sample is added, and then the reaction cup is placed in the reaction cup groove. The stirring system was automatically started and the reaction stirred for 10 seconds. The photoelectric control system controls the light source to be turned on, the reaction system is irradiated, the detection module records the detected optical signal and transmits the signal to the data collection and processing system. The control software automatically distinguishes whether the reagent being detected is a latex enhanced immunoturbidimetric reagent or a homogeneous chemiluminescent immunoassay reagent according to the selected detection item. If the reagent is a latex enhanced immunoturbidimetric reagent, the scattering signal is received and processed; in the case of homogeneous chemiluminescent reagents, the chemiluminescent signal is collected for processing. The time point and time span of collection of the optical signal are set according to the corresponding properties of the selected reagent. The detection time is generally about 3 to 5 minutes. And when the detection is finished, the operation interface prompts the completion of the detection. And the system software calculates the concentration of the object to be detected in the detection sample according to a pre-stored calibration curve of the corresponding reagent, and displays the result on a display or outputs the result to a printer.

Claims (8)

1. A real-time detection system of homogeneous immunoassay is characterized in that the real-time detection system comprises a light source, a photoelectric control system, a stirring system, a temperature control system, a reaction cup, a detection module and a data collection and processing system; the detection module comprises two or more light sensors; a lens or an optical filter is arranged on the optical sensor; the data collecting and processing system is connected with the detection module and is used for collecting optical signals and converting the optical signals into digital signals; the light source is used for generating light to irradiate a reaction system in the reaction cup; the detection module comprises a light path front detection module and a light path vertical direction detection module, and an optical sensor of the light path front detection module is used for receiving transmitted or scattered light; the optical sensor of the optical path vertical direction detection module is used for receiving a chemiluminescence signal; the optical sensor of the optical path front detection module is arranged right in front of the incident optical path or forms a certain included angle with the incident optical path; the included angle is an acute angle, a right angle or an obtuse angle; the detection system is also provided with a reaction cup groove, and the reaction cup groove is used for placing a reaction cup; the light-transmitting part of the reaction cup is made of transparent materials; the point-of-care assay system is adapted to be compatible with the use of latex turbidimetric immunoassay reagents and homogeneous luminescent immunoassay reagents.

2. The point-of-care detection system of claim 1, wherein the light source is a single light source or multiple light sources.

3. The point-of-care detection system of claim 2, wherein the light source is a halogen lamp, an LED, or a laser.

4. The real-time detection system as claimed in claim 3, wherein the light source emits light with a wavelength within the range of 400-750 nm.

5. The point-of-care assay system of claim 1, wherein the reagents in the cuvette are latex turbidimetric immunoassay reagents or homogeneous luminescent immunoassay reagents.

6. The in-situ detection system as claimed in claim 1, wherein the detection system comprises a module for detecting chemiluminescence with wavelength of 520-620nm and a module for detecting transmission or scattering light with wavelength of 400-750 nm.

7. The real-time detection system as claimed in claim 6, wherein a filter allowing passage of light waves with wavelength 520-620nm is disposed in front of the light measuring lens of the detection module for detecting chemiluminescence, and a photomultiplier is disposed for amplifying the light signal.

8. The point-of-care testing system of claim 1, wherein said stirring system is a magnetic stirring system.

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