Blood or Tissue Fluid Collection, Processing Chip, Detection Device and Method

Abstract

The present invention discloses a blood or tissue fluid collection and processing chip, a detection device and a method. Among them, the blood or tissue fluid collection and processing chip includes a human blood or tissue fluid collection mechanism and a human blood or tissue fluid in vitro processing mechanism. In the human blood or tissue fluid in vitro processing mechanism, there are single or multiple binding structures, a test chamber and a quality control chamber. In the binding structure, there are substances labeled with a labeling substance and capable of binding to a substance to be detected, and substances labeled with a labeling substance and capable of binding to a quality control substance. In the test chamber, there is a substance fixedly bound to the substance to be detected, and in the quality control chamber, there is a quality control substance fixedly bound. The detection device includes the blood or tissue fluid collection and processing chip as described above, and further includes an optical signal detection module, a data analysis and processing module, a display module, a control panel and a communication module, and can realize the collection, processing and detection of self-antibodies.

Description

Technical Field

[0001] The present invention relates to the technical field of pathogen detection, particularly to the technical field of blood or tissue fluid collection and processing chips, detection devices and methods.

Background Art

[0002] There are mainly three ways to detect whether a pathogen invades the human body, namely nucleic acid detection, antigen detection and antibody detection. Among them, nucleic acid detection determines whether a pathogen infection has occurred by detecting the pathogen nucleic acid in various body fluid specimens or tissue specimens through molecular biology techniques (such as real-time fluorescence polymerase chain reaction, isothermal amplification, in situ hybridization and sequencing, etc.). It has advantages such as high sensitivity, good specificity and strong reliability, and is mainly used for early diagnosis, prevention and control, prognosis judgment and efficacy monitoring of diseases. However, although nucleic acid detection can detect patients in the window period and thus discover infected persons as early as possible, it has relatively high requirements for detection equipment or platforms (high-sensitivity RT-PCR instruments are expensive), high requirements for the cleanliness of the laboratory and operators, and takes a long time. Considering the situation of sample transportation and sample backlog, it usually takes 4 to 8 hours to obtain the test report results. When the body is infected by various pathogens, it may lead to the presence of pathogen antigens in the blood, and antigen detection can diagnose whether the patient is infected by the pathogen. Compared with nucleic acid detection, antigen detection is faster and more convenient to operate, but has relatively low accuracy and is generally used in the early stage of infection. Antibody detection is to detect the immune response of the human body after being infected by a pathogen. Collecting relevant body fluids such as blood or tissue fluid is required for antibody detection. Compared with nucleic acid detection and antigen detection, professional medical staff are needed for collecting test specimens, and the process requires standardization, which is not conducive to grass-roots laboratories or individuals for self-detection. Moreover, the antibody content and status, such as antibodies like IgM, IgG and IgA, can clearly reflect the state of infection and the immune or defense response of the human body to the corresponding pathogen. Detecting antibodies can play a role in tracking and guiding the judgment of the course of relevant diseases and the treatment of diseases, which cannot be achieved by nucleic acid detection or antigen detection.

[0003] Currently, antibody detection is usually carried out by trained medical staff at fixed points for the collectors, and the whole process is limited by the professionalism of the venue, instruments and personnel, and there is also a risk of cross-infection. A rapid and accurate self-detection device and method for pathogen antibodies are very important for both the prognosis and treatment of infected persons with this pathogen and the evaluation of the effectiveness of vaccines against this pathogen.

Summary of the Invention

[0004] The object of the present invention is to solve the problems in the existing antibody detection technology, and to provide a blood or tissue fluid collection and processing chip, a detection device and a method, which can realize the collection, processing and detection of self-pathogen antibodies, and have the advantages of simple operation, accurate quantification, high sensitivity, short detection time and low cost. The collection and processing chip and its antibody detection device can be mass-produced and popularized, and are more suitable for individuals to monitor their own antibody production status.

[0005] To achieve the above object, the present invention provides a blood or tissue fluid collection and processing chip, which includes a human blood or tissue fluid collection mechanism and a human blood or tissue fluid external processing mechanism. The human blood or tissue fluid collection mechanism includes a single hollow microneedle or a microneedle array composed of multiple hollow microneedles arranged in an array. The human blood or tissue fluid external processing mechanism is provided with one or more binding structures, a test chamber and a quality control chamber. The binding structure is a microchannel and / or a binding chamber. One end of the binding structure is connected to the needle opening of the single or multiple hollow microneedles, and the other end is sequentially connected to the test chamber and the quality control chamber. The binding structure is loaded with a substance labeled with a labeling substance and capable of binding to the substance to be detected, and a substance labeled with a labeling substance and capable of binding to the quality control substance. When the collected blood or tissue fluid passes through the binding structure, the substance to be detected can fully bind to the above-labeled substance. A substance capable of binding to the substance to be detected is fixedly bound in the test chamber, and a quality control substance is fixedly bound in the quality control chamber.

[0006] Preferably, the length of the hollow microneedle is 20-200 μm.

[0007] Preferably, the number of the hollow microneedles is a single one or an array of multiple ones.

[0008] Preferably, the hollow microneedle can be inserted into an appropriate part of the human body (such as the abdomen and the inner side of the arm, etc.) and penetrate the epidermal layer.

[0009] Preferably, the hollow microneedle extracts microliters of blood or tissue fluid from under the epidermal layer by capillary action, pressure difference or a silicon microelectrostatic motor connected thereto.

[0010] Preferably, the tip of the hollow microneedle is sealed with a biofusion film, and the biofusion film can be dissolved by blood or tissue fluid after the hollow microneedle penetrates into the body.

[0011] Preferably, the human blood or tissue fluid external processing mechanism includes a substrate and a transparent cover plate. The binding structure, the test chamber and the quality control chamber are all arranged on the top of the substrate. The transparent cover plate is installed on the top surface of the substrate to enclose the binding structure, the test chamber and the quality control chamber and form a vacuum environment isolated from the outside or a closed environment filled with non-oxygen gas.

[0012] Further, the non-oxygen gas is nitrogen.

[0013] Further, the substrate is made of a polymer material, a pure metal material, an alloy material or a non-metal material.

[0014] Still further, the polymer material is PDMS or polystyrene, etc., the pure metal material is tungsten or titanium, etc., and the non-metal material is a carbon-based material, etc.

[0015] Further, the size of the microchannel is in the sub-micron to micron scale.

[0016] Further, the microchannel is meandering.

[0017] Further, the transparent cover plate is transparent inorganic glass, transparent organic glass or an optical organic film.

[0018] Still further, both the front and back surfaces of the transparent cover plate and the bottom surfaces of the test chamber and the quality control chamber are coated with an optical anti-reflection inorganic material coating.

[0019] Preferably, proteins can be immobilized or crosslinked on the walls of the test chamber and the quality control chamber.

[0020] Further, the function of immobilizing or crosslinking proteins is provided by the substrate itself or achieved through functional groups such as carboxyl and amino groups.

[0021] Further, the groups capable of immobilizing and crosslinking proteins are carboxyl or amino groups, etc.

[0022] Still further, the post-processing is coating a functional coating.

[0023] Preferably, the substance to be detected is human IgG antibody and / or human IgM antibody of the pathogen to be detected.

[0024] Preferably, the substance capable of binding to the substance to be detected is an antibody or an antigen.

[0025] Further, the antibody is anti-human IgG antibody and / or anti-human IgM antibody, and the antigen is natural antigen and / or recombinant antigen.

[0026] Still further, when at least two different substances capable of binding to the substance to be detected are loaded in the test chamber at the same time, the various substances capable of binding to the substance to be detected are respectively loaded in different test chambers or respectively loaded in different parts of the same test chamber.

[0027] Preferably, the labeling substance is one or a combination of several of colloidal gold, latex, fluorescent microspheres or fluorescein.

[0028] Preferably, the quality control substance is biotin, and the substance capable of binding to the quality control substance is avidin.

[0029] Preferably, the human blood or tissue fluid collection mechanism and the human blood or tissue fluid in vitro processing mechanism are integrated into one body and jointly form a microneedle and microfluidic integrated chip, or are two independent interconnected mechanisms.

[0030] The detection device includes the blood or tissue fluid collection and processing chip as described above, and also includes an optical signal detection module, a data analysis and processing module, a display module, a control panel, and a communication module. The data analysis and processing module is electrically connected to the optical signal detection module, the display module, the control panel, and the communication module respectively. The optical signal detection module can detect the optical signals of the test chamber and the quality control chamber, convert the optical signals into electrical signals, and transmit them to the data analysis and processing module. The data analysis and processing module can compare and analyze the electrical signals with the inventory data in the data center and use the communication module for data transmission.

[0031] Preferably, the optical signal detection module is a spectrometer or a spectrophotometer.

[0032] Furthermore, the optical signal detection module is an integrated optical sensor, photometric measurement, and amplifier of Analog Devices, such as series modules like ADPD10, 14, 17, 18X, 22XX, etc.

[0033] Preferably, the data analysis and processing module is the SH99F100 16-bit DSP + enhanced 8051 dual-core controller of Winbond Electronics.

[0034] Preferably, the display module is an optoelectronic thin-film touch display screen, which can also be used as a control panel.

[0035] Furthermore, the display module is, for example, H035045QO-54 of Hongxian Weiye.

[0036] Preferably, the communication module is a Bluetooth communication module, a 4G communication module, a 5G communication module, or other wireless communication modules.

[0037] Furthermore, the Bluetooth communication module is TTC HY-40R204P of Shengren Technology or MS50SFA of Yunli Wuli, etc.; the 4G communication module is ML303 of China Mobile or EC04-SGC of Ebyte Electronics, etc.; the 5G communication module is RG200U-CN of Quectel Wireless Solutions; the other wireless communication modules are E840-TTL-NB03 of Ebyte Electronics or BC28JC-02-B5 of Quectel Wireless Solutions.

[0038] The above modules are all mature products in the market and can be optimized and selected according to the actual design.

[0039] Preferably, it further includes an environmental temperature detection module. The environmental temperature detection module can detect whether the blood or tissue fluid collection and processing chip is in an extreme environmental temperature, convert the temperature signal into an electrical signal and transmit it to the data analysis and processing module, so as to correct the influence of the extreme test environmental temperature on the measurement data to obtain an accurate antibody concentration.

[0040] Furthermore, the environmental temperature detection module is BSY0-103IOV1 of BYD Electronics.

[0041] The detection method uses the pathogen antibody self-test device as described above and includes the following steps:

[0042] a) Sampling: Insert the hollow micro-needle into the epidermis so that human blood or tissue fluid flows through the binding structure, the test chamber and the quality control chamber in sequence;

[0043] b) Detection: Use the optical signal detection module to simultaneously detect the optical signals in the test chamber and the quality control chamber. First, distinguish human tissue fluid or blood by the optical signal in the test chamber, and at the same time judge whether there is a substance to be detected in the human body by whether the optical signal in the test chamber is generated. Then, obtain the concentration of the substance to be detected in the human body by the intensity of the optical signal in the test chamber and the intensity of the optical signal in the quality control chamber.

[0044] In addition, it should be emphasized that the blood or tissue fluid collection and processing chip, the detection device and the method of the present invention can also be used to detect non-antibody proteins or small molecule substances in blood or tissue fluid, etc.

[0045] Advantages of the present invention:

[0046] 1) By setting up a human blood or tissue fluid collection mechanism and a human blood or tissue fluid external processing mechanism to jointly form a blood or tissue fluid collection and processing chip, since the hollow micro-needle only pierces the epidermis, there is no pain, the overall operation is simple, the sample collection error rate is low, and the collection and processing chip is a disposable consumable. It can be processed and manufactured by using the mature micron-scale large-scale integrated circuit production technology and equipment in the microelectronics industry, and its large-scale production cost will not be higher than 10 yuan. It meets the cost requirements for personal or family use;

[0047] 2) The blood or tissue fluid collection and processing chip disclosed by the present invention and its supporting detection and analysis device can be easily loaded and unloaded and connected. The antibody detection results will be displayed on a separate display and / or personal mobile phone. The supporting detection and analysis device includes an optical signal detection module, a data analysis and processing module, a display module, a control panel, and a communication module. Furthermore, rapid quantitative detection of pathogen antibodies can be achieved by utilizing the change of optical signals, enabling pathogen-infected individuals, such as those with mild symptoms or asymptomatic confirmed cases of the novel coronavirus, to directly conduct self-antibody detection, and then autonomously analyze the condition according to personal characteristics and physical conditions. The data can also be uploaded to relevant medical institutions or disease control departments to obtain remote medical support, thereby making it possible for pathogen-infected individuals to accurately predict the development of their own conditions and for vaccine recipients to accurately monitor the growth of their own pathogen antibodies;

[0048] 3) By integrating the human blood or tissue fluid collection mechanism and the human blood or tissue fluid external processing mechanism into one body to jointly form a micro-needle and microfluidic integrated chip, the present invention can use a biological fusion film to seal the tip of the hollow micro-needle. At the same time, a transparent cover plate is provided on the top surface of the substrate to simultaneously seal the needle opening of the hollow micro-needle, the micro-channel, the test cavity, and the quality control cavity, and then vacuum or inflation treatment is carried out, so that the blood or tissue fluid collection and processing chip forms an environment isolated from the outside world, effectively avoiding the contact of blood or tissue fluid with external air during extraction and processing, and reducing the influence of factors such as environmental humidity, atmosphere, and air pressure on the detection results;

[0049] 4) By adding an environmental temperature detection module, the present invention further reduces the influence of extreme temperature conditions on the test results and improves the accuracy of the detection results.

[0050] The features and advantages of the present invention will be described in detail through embodiments in conjunction with the drawings.

Description of the Drawings

[0051] Figure 1 is a three-dimensional structural schematic diagram of the blood or tissue fluid collection and processing chip of Embodiment 1;

[0052] Figure 2 is a front view cross-sectional view of the blood or tissue fluid collection and processing chip of Embodiment 1;

[0053] Figure 3 is a top view of the blood or tissue fluid collection and processing chip of Embodiment 1 after removing the transparent cover plate;

[0054] Figure 4 is a top view of the blood or tissue fluid collection and processing chip of Embodiment 2 after removing the transparent cover plate.

[0055] In the figure: 1 - human blood or tissue fluid collection mechanism, 2 - human blood or tissue fluid external processing mechanism, 21 - substrate, 211 - microchannel, 212 - test chamber, 213 - quality control chamber, 214 - binding chamber, 22 - transparent cover plate.

Specific Embodiment

[0056] Embodiment 1:

[0057] Taking the detection of novel coronavirus as an example, referring to Figure 1 , Figure 2 and Figure 3 , the blood or tissue fluid collection and processing chip of the present invention includes a human blood or tissue fluid collection mechanism 1 and a human blood or tissue fluid external processing mechanism 2. The human blood or tissue fluid collection mechanism 1 includes a single hollow micro needle. In the human blood or tissue fluid external processing mechanism 2, there is a single microchannel 211, a test chamber 212 and a quality control chamber 213. One end of the microchannel 211 is connected to the needle opening of the hollow micro needle, and the other end is sequentially connected to the test chamber 212 and the quality control chamber 213. The microchannel 211 is loaded with novel coronavirus antigen S protein and / or N protein labeled with a labeling substance and avidin labeled with a labeling substance. The test chamber 212 is fixedly loaded with anti-human immunoglobulin antibody, and the quality control chamber 213 is fixedly loaded with biotin.

[0058] The tip of the hollow micro needle is sealed with a biofusion film, and the biofusion film can be melted by blood or tissue fluid after the hollow micro needle penetrates into the body.

[0059] The human blood or tissue fluid external processing mechanism 2 includes a substrate 21 and a transparent cover plate 22. The microchannel 211, the test chamber 212 and the quality control chamber 213 are all arranged on the top of the substrate 21. The transparent cover plate 22 is installed on the top surface of the substrate 21 to enclose the microchannel 211, the test chamber 212 and the quality control chamber 213 and form a vacuum environment isolated from the outside.

[0060] An anti-reflection coating is coated on the bottom surface of the test chamber 212. The surface of the anti-reflection coating is rich in carboxyl groups, or is treated with a nano-coating to be rich in carboxyl groups. The anti-human immunoglobulin antibody can be cross-linked and fixed through the carboxyl groups.

[0061] An anti-reflection coating is coated on the bottom surface of the quality control chamber 213, and biotin is cross-linked on the surface of the anti-reflection coating.

[0062] Anti-reflection coatings are coated on both surfaces of the transparent cover plate 22.

[0063] The anti-human immunoglobulin antibody is mouse anti-human IgG antibody and / or mouse anti-human IgM antibody.

[0064] When both the mouse anti - human IgG antibody and the mouse anti - human IgM antibody are loaded in the test chamber 212, the mouse anti - human IgG antibody and the mouse anti - human IgM antibody are respectively loaded at different positions in the same test chamber 212.

[0065] The labeling substance is one or a combination of several of colloidal gold, latex, fluorescent microspheres or fluorescein.

[0066] The human blood or tissue fluid collection mechanism 1 and the human blood or tissue fluid in - vitro processing mechanism 2 are integrated into one body and jointly form a microneedle and microfluidic integrated chip, or are two independent interconnected mechanisms.

[0067] The detection device includes the blood or tissue fluid collection and processing chip as described above, and also includes an optical signal detection module, a data analysis and processing module, a display module, a control panel and a communication module. The data analysis and processing module is electrically connected to the optical signal detection module, the display module, the control panel and the communication module respectively. The optical signal detection module can detect the optical signals of the test chamber 212 and the quality control chamber 213, convert the optical signals into electrical signals and transmit them to the data analysis and processing module. The data analysis and processing module can compare and analyze the electrical signals with the inventory data in the data center and use the communication module for data transmission.

[0068] The detection method uses the detection device as described above and includes the following steps:

[0069] a) Sampling: Insert the hollow microneedle into the human body and penetrate the epidermal layer, so that the blood or tissue fluid flows through the microchannel 211, the test chamber 212 and the quality control chamber 213 in sequence.

[0070] b) Detection: Use the optical signal detection module to simultaneously detect the optical signals in the test chamber 212 and the quality control chamber 213. First, distinguish human tissue fluid or blood by the optical signal in the test chamber 212, and at the same time judge whether there is a substance to be detected in the human body by whether the optical signal in the test chamber 212 is generated. Then, obtain the concentration of the substance to be detected in the human body by the intensity of the optical signal in the test chamber 212 and the intensity of the optical signal in the quality control chamber 213.

[0071] The working process of the present invention:

[0072] First, the needle of the pathogen antibody self-test device, which has been installed with a blood or tissue fluid collection and processing chip, is inserted into the human body and passes through the epidermis, so that the blood or tissue fluid flows gradually toward the test cavity 212 along the microchannel 211 under the guidance of capillary force from the needle opening of the hollow microneedle. During the flow process, if there is new crown antibody IgM or IgG in the blood or tissue fluid, it will be fully combined with the new crown antigen S protein and / or N protein labeled with the labeled substance loaded in the microchannel 211 due to the laminar flow and Dean effect unique to the microchannel 211, and then reach the test cavity 212 together. When the sample flows to the test cavity 212, the flow rate will be significantly slowed down, and after the antibody in the sample is combined with the labeled antigen, it will be captured by the mouse anti-human IgG antibody and / or mouse anti-human IgM antibody fixed in the test cavity 212 by chemical cross-linking, and produce color change or generate fluorescence under the excitation of incident light, and then the change intensity is detected by a spectrophotometer or spectrometer. In addition, the components that are not captured will continue to flow with the whole body fluid until they flow to the quality control cavity 213. At this time, the biotin fixed by chemical cross-linking in the quality control cavity 213 will capture the avidin labeled with the labeling substance, thereby causing a color change or generating fluorescence under the excitation of incident light, and then the intensity of the change is detected by a spectrophotometer or spectrometer to form a comparison. Figure 1 Indicated by the arrow direction.

[0073] Subsequently, the data analysis and processing module compares and analyzes the detected data with the data model of virus antibody levels and infection symptoms in the data center, thereby providing accurate prediction results of the development of infection symptoms. In addition, the data analysis and processing module can also transmit the detection data to the mobile phone APP or send it to the medical / disease control center through the communication module to obtain professional medical support.

[0074] Embodiment 2:

[0075] The microchannel 211 is replaced by the combination cavity 214 , and the rest is the same as in the first embodiment.

[0076] The above embodiments are intended to illustrate the present invention, not to limit the present invention. Any solution that is a simple transformation of the present invention belongs to the protection scope of the present invention.

Claims

1. Blood or tissue fluid collection and processing chip, Characterized in that: It includes a human blood or tissue fluid collection mechanism (1) and a human blood or tissue fluid in vitro processing mechanism (2). The human blood or tissue fluid collection mechanism (1) includes a single hollow micro needle or a micro needle array composed of multiple hollow micro needles arranged. In the human blood or tissue fluid in vitro processing mechanism (2), there are single or multiple binding structures, a test chamber (212) and a quality control chamber (213). The binding structure is a combination of a microchannel (211) or a microchannel (211) and a binding chamber (214). One end of the binding structure is connected to the needle opening of a single or multiple hollow micro needles, and the other end is sequentially connected to the test chamber (212) and the quality control chamber (213). The binding structure is loaded with a substance labeled with a labeling substance and capable of binding to the substance to be detected and a substance labeled with a labeling substance and capable of binding to the quality control substance. In the test chamber (212), a substance capable of binding to the substance to be detected is fixedly bound. In the quality control chamber (213), a quality control substance is fixedly bound. The tip of the hollow micro needle is sealed with a biofusion film, and the biofusion film can be melted by blood or tissue fluid after the hollow micro needle penetrates into the body. The human blood or tissue fluid in vitro processing mechanism (2) includes a substrate (21) and a transparent cover plate (22). The binding structure, the test chamber (212) and the quality control chamber (213) are all arranged on the top of the substrate (21). The transparent cover plate (22) is installed on the top surface of the substrate (21) to enclose the binding structure, the test chamber (212) and the quality control chamber (213) and form a vacuum environment isolated from the outside. The microchannel (211) is serpentine. The front and back surfaces of the transparent cover plate (22) and the bottom surface of the test chamber (212) and the quality control chamber (213) are all coated with an optical anti-reflection inorganic material coating. When at least two different substances capable of binding to the substance to be detected are simultaneously loaded in the test chamber (212), each of the substances capable of binding to the substance to be detected is respectively loaded in different test chambers (212) or respectively loaded in different parts of the same test chamber (212). The hollow micro needle extracts microliters of blood or tissue fluid from under the epidermal layer through capillary action or pressure difference.

2. The blood or tissue fluid collection and processing chip according to claim 1, Characterized in that: The substance capable of binding to the substance to be detected is an antibody or an antigen.

3. The blood or tissue fluid collection and processing chip according to claim 2, Characterized in that: The antibody is an anti-human IgG antibody and / or an anti-human IgM antibody, and the antigen is a natural antigen and / or a recombinant antigen.

4. The blood or tissue fluid collection and processing chip according to claim 1, Characterized in that: The labeling substance is one or a combination of several of colloidal gold, latex, fluorescent microspheres or fluorescein.

5. The blood or tissue fluid collection and processing chip according to claim 1, Characterized in that: The human blood or tissue fluid collection mechanism (1) and the human blood or tissue fluid external processing mechanism (2) are integrated into one body and jointly form a microneedle and microfluidic integrated chip, or are two independent mechanisms connected to each other.

6. Detection device, characterized in that: It includes the blood or tissue fluid collection and processing chip according to any one of claims 1 to 5, and further includes an optical signal detection module, a data analysis and processing module, a display module, a control panel and a communication module. The data analysis and processing module is electrically connected to the optical signal detection module, the display module, the control panel and the communication module respectively. The optical signal detection module can detect the optical signals of the test chamber (212) and the quality control chamber (213), convert the optical signals into electrical signals and transmit them to the data analysis and processing module. The data analysis and processing module can compare and analyze the electrical signals with the inventory data in the data center and use the communication module for data transmission.

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