CN109856398B - Vaccine detection device and detection method thereof - Google Patents
Vaccine detection device and detection method thereof Download PDFInfo
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- CN109856398B CN109856398B CN201910100705.3A CN201910100705A CN109856398B CN 109856398 B CN109856398 B CN 109856398B CN 201910100705 A CN201910100705 A CN 201910100705A CN 109856398 B CN109856398 B CN 109856398B
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Abstract
The application provides a vaccine detection device and a detection method thereof. The vaccine detection device is used for detecting the effectiveness of a vaccine, the vaccine is an inactivated type or attenuated type single vaccine, and the vaccine detection device comprises: the detection channel comprises a drawing end and a driving end, a reaction area and a detection area are sequentially arranged in the detection channel from the drawing end to the driving end, a first antibody is arranged in the reaction area, the first antibody can be specifically combined with an inactivated or attenuated pathogen in a vaccine to be detected, the first antibody in the reaction area is labeled by colloidal gold, and the first antibody is arranged in the detection area; and the driving assembly is positioned at the driving end of the detection channel and is used for driving the vaccine to enter the detection channel from the drawing end of the detection channel and flow in the driving direction. The vaccine detection method adopts the vaccine detection device to detect. The vaccine detection device can be used for detecting the effectiveness of the vaccine quickly.
Description
Technical Field
The application relates to a vaccine detection device and a detection method thereof.
Background
Inactivated or attenuated unions are prepared by inactivating pathogens (viruses or bacteria) or by subtracting the virulent form of the pathogen to humans. The vaccine retains the antigenic property of pathogen, and can induce specific immune response of body, so that human can obtain immune protection to the pathogen. The vaccines are harsh in storage conditions, generally need to be stored under the conditions of light protection and low temperature, and are easy to lose efficacy in the transportation and storage processes. Meanwhile, a large number of vaccine counterfeiting events are reported in China, so that people can hardly judge whether the inoculated vaccine is effective or not.
At present, the effectiveness detection of vaccine detection is carried out by depending on a large-scale detection mechanism. This detection method is time and labor consuming. And the amount of single vaccine consumed in detection is large, while the dose of the common vaccine is very small, and the effective vaccination dose of the vaccine can not be reached after the detection. Therefore, at present, vaccine detection can only be stopped at a spot check stage, and whether the vaccine is effective or not can not be detected before inoculation.
Disclosure of Invention
The invention provides a vaccine detection device and a detection method thereof.
In order to achieve the above purpose, the embodiment of the present invention provides a vaccine detection apparatus. The vaccine detection device is used for detecting the effectiveness of a vaccine, wherein the vaccine is an inactivated type or attenuated type single vaccine, and the vaccine detection device comprises:
the detection device comprises a main body, wherein a detection channel is arranged in the main body, the detection channel comprises a drawing end and a driving end, a reaction area and a detection area are sequentially arranged in the detection channel from the drawing end to the driving end, a first antibody is arranged in the reaction area, the first antibody can be specifically combined with an inactivated or attenuated pathogen in the vaccine to be detected, the first antibody in the reaction area is labeled by colloidal gold, and the detection area is provided with the first antibody;
the driving assembly is positioned at the driving end of the detection channel and is used for driving the vaccine to enter the detection channel from the drawing end of the detection channel and flow in the driving direction.
Optionally, the first antibody labeled with the colloidal gold in the reaction region is spotted on the tube wall of the detection channel by electrostatic adsorption;
and/or the first antibody of the detection area is spotted on the tube wall of the detection channel by an embedding method.
Optionally, the driving assembly includes a piston and a piston rod connected to each other, and the piston is clamped at the driving end of the detection channel and can be driven by the piston rod to reciprocate along the axial direction of the detection channel.
Optionally, the vaccine detection apparatus further includes a microfluidic component, the microfluidic component is fixed to the main body, and the microfluidic component is configured to limit a moving distance of the driving component along an axial direction of the detection channel.
Optionally, the microfluidic component includes a screw nut and a screw rod matched with the screw nut in a threaded manner, the screw nut and the main body are relatively fixedly arranged, and the screw rod is connected with the driving component and drives the driving component to move along the axial direction of the detection channel in a reciprocating manner.
Optionally, the vaccine detection device further comprises a drawing part, the drawing part is arranged at the drawing end of the detection channel, the drawing part is a needle head, the needle head is fixed on the fixing seat, the fixing seat is connected with the main body, and the needle head is communicated with the drawing end of the detection channel.
Optionally, a control area is further disposed in the detection channel, the control area is located at one end of the detection area, which is far away from the reaction area, the control area is provided with a second antibody, and the second antibody can specifically bind to the first antibody.
Optionally, the second antibody of the control zone is spotted on the tube wall of the detection channel by an embedding method.
Optionally, the inner surface of the tube wall of the detection channel except the reaction region, the detection region, and the control region is subjected to surface hydrophobic treatment.
The embodiment also provides a vaccine detection method, which uses the vaccine detection device to detect, and the vaccine detection method includes:
enabling the vaccine to be detected to enter the detection channel from the suction end of the detection channel through the driving assembly, and sequentially passing through the reaction region and the detection region in the driving direction;
when the inactivated or attenuated pathogen in the vaccine to be detected can be specifically combined with the first antibody marked by the colloidal gold in the reaction area and reacts with the first antibody in the detection area, so that the color of the detection area is displayed by the colloidal gold, the vaccine to be detected is an effective vaccine; when the inactivated or attenuated pathogen in the vaccine to be detected cannot react with the first antibody marked by the colloidal gold, the detection area does not display color, and the vaccine to be detected is an ineffective vaccine.
Optionally, a control area is further disposed in the detection channel, the control area is located at one end of the detection area away from the reaction area, the control area is provided with a second antibody, and the second antibody can specifically bind to the first antibody;
the detection method of the vaccine further comprises the following steps: when the first antibody marked by the colloidal gold can react with the second antibody in the control area, so that the control area displays color through the colloidal gold, the detection is effective; and if the first antibody marked by the colloidal gold cannot react with the second antibody in the control area, the control area does not display color, and the detection is invalid.
In the vaccine detection device and the vaccine detection method of the embodiment, the effectiveness of the vaccine can be rapidly detected by setting the overall structure of the vaccine detection device.
Drawings
FIG. 1 is a structural view of an exemplary embodiment vaccine testing device.
Fig. 2 is an internal structural view of an exemplary embodiment vaccine testing device.
Description of the reference numerals
Tapping end 111
Drive end 112
Piston 21
Piston rod 22
Screw adjusting nut 31
Rotary adjusting screw 32
Needle 40
Fixed seat 50
Detailed Description
Reference will now be made in detail to the exemplary embodiments, examples of which are illustrated in the accompanying drawings. When the following description refers to the accompanying drawings, like numbers in different drawings represent the same or similar elements unless otherwise indicated. The embodiments described in the following exemplary embodiments do not represent all embodiments consistent with the present application. Rather, they are merely examples of apparatus consistent with certain aspects of the present application, as detailed in the appended claims.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. Unless defined otherwise, technical or scientific terms used herein shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The use of the terms "a" or "an" and the like in the description and in the claims of this application do not denote a limitation of quantity, but rather denote the presence of at least one. The word "comprising" or "comprises", and the like, means that the element or item listed as preceding "comprising" or "includes" covers the element or item listed as following "comprising" or "includes" and its equivalents, and does not exclude other elements or items. The terms "connected" or "coupled" and the like are not restricted to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "plurality" includes two, and is equivalent to at least two. As used in this specification and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It should also be understood that the term "and/or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.
As will be understood with reference to fig. 1 and 2, the present embodiment provides a vaccine testing device. The vaccine detection device is used for detecting the effectiveness of a vaccine, wherein the vaccine is an inactivated type or attenuated type single vaccine, and the vaccine detection device comprises a main body 10 and a driving component 20.
The detection channel 11 is arranged in the main body 10, the detection channel 11 includes a drawing end 111 and a driving end 112, and the detection channel 11 is provided with a reaction region 113 and a detection region 114 in sequence from the drawing end 111 to the driving end 112. The width of the detection channel 11 is 0.3-2 mm, and the cross-sectional shape may be rectangular, circular, or other shapes. The tapping end 111 of the detection channel 11 is provided with graduation marks 116, and the graduation marks 116 are marked with 1-5 microliter graduations.
The reaction region 113 is provided with a first antibody, the first antibody can specifically bind to an inactivated or attenuated pathogen in the vaccine to be detected, and the first antibody of the reaction region 113 is labeled with colloidal gold. The detection zone 114 is provided with the primary antibody. In this embodiment, the first antibody labeled with the colloidal gold in the reaction region 113 is spotted on the tube wall of the detection channel 11 by electrostatic adsorption; the first antibody of the detection region 114 is spotted on the tube wall of the detection channel 11 by an embedding method. The first antibody labeled with the colloidal gold is applied by electrostatic adsorption, so that the first antibody labeled with the colloidal gold can be relatively easily separated from the reaction region 113 and moved to the detection region 114. The first antibody is spotted by an embedding method, so that the first antibody can be more firmly fixed on the detection region 114. The immunoglobulin is fixed on the substrate by adopting an embedding method, has the characteristics of stable sample application, no enzyme separation at low flow rate and the like, and simultaneously leaves a reaction region for the antigen and the antibody and generates a detection result in the detection region 114. Specifically, in this embodiment, the first antibody of the PDDA immobilized detection region 114 is used for explanation, and the specific operations are as follows: and (3) carrying out ultraviolet irradiation on the sample application region, soaking the sample application region with acrylic acid to graft the sample application region with acrylic acid, simultaneously mixing the sample application first antibody with PDDA, then carrying out sample application on the mixture to the acrylic acid soaking region, adsorbing the PDDA and the acrylic acid together, simultaneously wrapping the first antibody by the net structure of the PDDA, and carrying out air drying by nitrogen wind to complete the sample application. Wherein PDDA is the English abbreviation of poly dimethyl diallyl ammonium chloride.
For the convenience of observation, the material of the tube wall of the detection channel 11 is transparent. And the main body 10 has a triangular structure, so that the detection result of the detection channel 11 can be observed more clearly.
The driving assembly 20 is located at the driving end 112 of the detection channel 11 and is used for driving the vaccine to enter the detection channel 11 from the drawing end 111 of the detection channel 11 and flow in the driving direction. In this way, the vaccine to be detected can be driven to flow in the detection channel 11 rapidly according to the driving direction by the driving component 20, so that the detection can be completed rapidly; meanwhile, the detection result can be directly observed by naked eyes in a mode of labeling the first antibody by the colloidal gold without other observation tools, so that the convenient and fast detection can be realized.
The driving assembly 20 includes a piston 21 and a piston rod 22 connected to each other, the piston 21 is clamped at the driving end 112 of the detection channel 11, and can be driven by the piston rod 22 to reciprocate along the axial direction of the detection channel 11. The piston rod 22 is driven by external force to make the piston 21 move back and forth, so as to provide driving force for the liquid in the detection channel 11.
The vaccine detection device further comprises a micro-fluidic component 30, the micro-fluidic component 30 is relatively fixedly arranged with the main body 10 and is located on one side of the driving component 20 far away from the driving end 112 of the detection channel 11, and the micro-fluidic component 30 is used for limiting the axial moving distance of the driving component 20 along the detection channel 11, so that the liquid absorption amount can be finely adjusted, and the purpose of controlling the use amount of the required vaccine to be detected in a small range is achieved. The amount of vaccine to be tested required in this example is very small, requiring only 2-5 microliters of vaccine. Specifically, the vaccine to be detected in this embodiment is a rabies virus vaccine, and usually each rabies virus vaccine (after reconstitution) is 0.5ml, and the rabies virus vaccine to be detected only accounts for 0.4% -1% of the total amount of the whole rabies virus vaccine, and does not affect rabies virus vaccination. Compared with the prior art that the rabies virus vaccine is detected by the detection test paper, the detection test paper needs more detection amount, and the dosage of the rabies virus vaccine is extremely small (mostly only 0.5ml), so that the rabies virus vaccine is not suitable for detection by the detection test paper.
The micro-fluidic component 30 comprises a rotary adjusting nut 31 and a rotary adjusting screw 32 in threaded fit with the rotary adjusting nut 31, the rotary adjusting nut 31 is relatively fixed with the main body 10, and the rotary adjusting screw 32 is connected with the driving component 20 and drives the driving component 20 to reciprocate along the axial direction of the detection channel 11. During detection, the screw nut 31 can be rotated to drive the screw rod 32 to slightly move along the axial direction of the detection channel 11, so that the piston 21 can slightly move along the axial direction of the detection channel 11, and the dosage of the required vaccine to be detected is limited within a small range.
Furthermore, a bottom cover 60 is arranged at one end of the screwing nut 31 far away from the driving component 20, an elastic element 70 is arranged between the bottom cover 60 and the screwing screw rod 32, two ends of the elastic element 70 are respectively connected with the bottom cover 60 and the screwing screw rod 32, and an acting force towards the drawing end 111 of the detection channel 11 can be always applied to the screwing screw rod 32 through the arrangement of the elastic element 70, so that when the screwing screw rod 32 rotates to move towards the direction far away from the drawing end 111 of the detection channel 11, the resistance can be increased, and the dosage of the vaccine to be detected entering the drawing end 111 of the detection channel 11 can be better controlled.
The vaccine detection device further comprises a drawing part, the drawing part is arranged at a drawing end 111 of the detection channel 11, the drawing part is a needle head 40, the needle head 40 is fixed on the fixing seat 50, the fixing seat 50 is connected with the main body 10, and the needle head 40 is communicated with the drawing end 111 of the detection channel 11. In this way, the vaccine to be tested can be conveniently taken out of the ampoule by providing the drawing part and directly enter the drawing end 111 of the test channel 11.
The detection channel 11 is further provided with a control area 115, the control area 115 is located at one end of the detection area 114 far away from the reaction area 113, and the control area 115 is provided with a second antibody which can be specifically combined with the first antibody. The second antibody of the control zone 115 is spotted on the wall of the detection channel 11 by an embedding method. Whether the first antibody in the reaction region 113 and the detection region 114 is effective can be detected by providing the control region 115, thereby ensuring the effectiveness of the detection.
The inner surface of the tube wall of the detection channel 11 except for the reaction region 113, the detection region 114, and the control region 115 is subjected to surface hydrophobic treatment. Thus, the liquid can move relatively quickly in the regions other than the reaction region 113, the detection region 114, and the control region 115, thereby further shortening the detection time.
The vaccine detection device of the embodiment can detect the effectiveness of the vaccine before vaccination, and judge whether the vaccine to be vaccinated is ineffective or false vaccine. The vaccine detection device of the embodiment is simple to operate, the detection time is short (within 5 minutes), the detection result is easy to judge, and professional personnel operation and expensive instruments and equipment are not needed.
Specifically, the vaccine detection apparatus of the present example was used to detect the effectiveness of a rabies virus vaccine by using a canine IgG as the first antibody and one of a rabbit anti-RV IgG, a goat anti-RV IgG, and a mouse anti-RV IgG as the second antibody, and herein the rabbit anti-RV IgG was used as the second antibody. Wherein IgG is an abbreviation for immunoglobulin G; RV is called Rabies virus in English and Rabies virus in Chinese. Canine IgG refers to canine immunoglobulin G; rabbit anti-RV IgG refers to an immunoglobulin produced by the rabbit immune system when inoculated with rabies virus immunoglobulin G; goat anti-RV IgG refers to an immunoglobulin produced by the immune system of a goat when inoculated with rabies virus immunoglobulin G; murine anti-RV IgG refers to an immunoglobulin produced by the murine immune system when inoculated with rabies virus immunoglobulin G.
The embodiment also provides a vaccine detection method, which uses the vaccine detection device to detect, and the vaccine detection method includes:
the vaccine to be detected enters the detection channel 11 from the drawing end 111 of the detection channel 11 through the driving assembly 20, and sequentially passes through the reaction area 113, the detection area 114 and the control area 115 according to the driving direction;
when the inactivated or attenuated pathogen in the vaccine to be detected can be specifically combined with the first antibody marked by the colloidal gold in the reaction region 113 to form pathogen-first antibody-colloidal gold, and can be reacted with the first antibody in the detection region 114 to form first antibody-pathogen-first antibody-colloidal gold, so that the detection region 114 displays color through the colloidal gold, the vaccine to be detected is an effective vaccine; when the inactivated or attenuated pathogen in the vaccine to be detected cannot react with the first antibody labeled by the colloidal gold, the detection area 114 does not display color, and the vaccine to be detected is an ineffective vaccine;
when the first antibody labeled by the colloidal gold can react with the second antibody in the control area 115, so that the control area 115 displays a color through the colloidal gold, the detection is valid; the first antibody labeled with the colloidal gold cannot react with the second antibody in the control area 115, and the control area 115 does not display color, so that the detection is invalid.
Specifically, when the method for detecting a rabies virus vaccine of this embodiment is used to detect the effectiveness of a rabies virus vaccine, the needle 40 of the vaccine detection apparatus of this embodiment is inserted into a liquid of a rabies virus vaccine (redissolution) to be inoculated, the turnnut 31 is adjusted, the piston 21 is driven to draw 3 microliters of the rabies virus vaccine to be detected (herein, referred to as a sample), and then the needle 40 is taken out of the vaccine. The adjustment of the turnnut 31 is continued to hold the drawn sample in position in the reaction area 113 for 2 minutes. The turn nut 31 is adjusted so that the sample moves to the position of the detection zone 114 and the control zone 115, respectively, for one minute each. Finally, the nut 31 is turned in the opposite direction to move the sample out of the detection channel 11.
When the vaccine is detected, the colors of the detection area 114 and the control area 115 are observed, if the detection area 114 is red, the sample is a positive sample, and the rabies virus vaccine is indicated to be an effective vaccine. The inactivated rabies virus in the positive sample is subjected to antigen-antibody specific binding with the colloidal gold-labeled canine IgG at the reaction area 113 to form virus-canine IgG-colloidal gold. When the test sample moves to the detection region 114, the virus-canine IgG-colloidal gold reacts again with the canine IgG immobilized on the detection region 114 to form canine IgG-virus-canine IgG-colloidal gold, so that the detection region 114 shows a red color.
If the detection area 114 does not show red color, the sample is a negative sample, which indicates that the rabies virus vaccine is an ineffective vaccine.
Then, the sample is moved to the position of the control region 115, and the colloidal gold-labeled dog IgG reacts with the rabbit anti-RVIgG to form rabbit anti-RV IgG-dog IgG-colloidal gold, so that the control region 115 shows red, which indicates that the colloidal gold-labeled dog IgG used in the detection is effective, i.e., the detection result is effective. If the control area 115 does not display red, the detection result is invalid.
If the detection area 114 does not display color, the sample is a negative sample, which indicates that the rabies virus vaccine is an ineffective vaccine. This is because there is no inactivated rabies virus or denaturation of rabies virus antigen protein, and thus the reaction region 113 cannot react with the canine IgG in the detection region 114, so that the detection region 114 does not show red color.
According to the vaccine detection device, the effectiveness of the vaccine can be rapidly detected by setting the integral structure of the vaccine detection device. Specifically, the vaccine detection device of the present embodiment achieves the following effective effects:
1. the invention combines the micro-fluidic technology, so that the sample size only needs 2-5 microliter of vaccine, and the subsequent use of the vaccine is not influenced.
2. The detection needs short time, the invention adopts the colloidal gold immunoreaction technology, the specific combination of the antibody and the antigen only needs a plurality of minutes, and the detection timeliness is kept.
3. The method does not need professional operation, and is effective, simple and feasible in vaccine judgment by observing the color development of the colloidal gold.
The above description is only exemplary of the present application and should not be taken as limiting the present application, as any modification, equivalent replacement, or improvement made within the spirit and principle of the present application should be included in the scope of protection of the present application.
Claims (11)
1. A vaccine testing device for testing the effectiveness of a vaccine, wherein the vaccine is an inactivated or attenuated univaccine, and the vaccine testing device comprises:
the detection device comprises a main body, wherein a detection channel is arranged in the main body, the pipe wall material of the detection channel is a transparent material, the detection channel comprises a drawing end and a driving end, a reaction area and a detection area are sequentially arranged in the direction from the drawing end to the driving end of the detection channel, a first antibody is arranged in the reaction area, the first antibody can be specifically combined with an inactivated or attenuated pathogen in the vaccine to be detected through antigen-antibody combination, the first antibody in the reaction area is labeled through colloidal gold, and the detection area is provided with the first antibody;
the driving assembly is positioned at the driving end of the detection channel and is used for driving the vaccine to enter the detection channel from the drawing end of the detection channel and flow in the driving direction.
2. The vaccine detection device according to claim 1, wherein the first antibody labeled with the colloidal gold in the reaction region is spotted on the tube wall of the detection channel by electrostatic adsorption;
and/or the first antibody of the detection area is spotted on the tube wall of the detection channel by an embedding method.
3. The vaccine detection apparatus according to claim 1, wherein the driving assembly includes a piston and a piston rod connected to each other, and the piston is engaged with the driving end of the detection channel and is capable of moving back and forth along the axial direction of the detection channel under the driving of the piston rod.
4. The vaccine detection apparatus according to claim 1, further comprising a microfluidic assembly fixedly disposed relative to the main body, the microfluidic assembly configured to limit a distance of movement of the driving assembly along an axial direction of the detection channel.
5. The vaccine detection apparatus according to claim 4, wherein the microfluidic assembly includes a screw nut and a screw rod threadedly engaged with the screw nut, the screw nut is fixed to the main body, and the screw rod is connected to the driving assembly and drives the driving assembly to reciprocate along the axial direction of the detection channel.
6. The vaccine testing device according to claim 1, further comprising a tapping component disposed at a tapping end of the testing channel; the drawing part is a needle head, the needle head is fixed on a fixed seat, the fixed seat is connected with the main body, and the needle head is communicated with the drawing end of the detection channel.
7. The vaccine detection device according to any one of claims 1 to 6, wherein a control zone is further provided in the detection channel, the control zone is located at an end of the detection zone away from the reaction zone, and the control zone is provided with a second antibody, and the second antibody is capable of specifically binding to the first antibody.
8. The vaccine detection apparatus of claim 7, wherein the second antibody of the control zone is spotted on the tube wall of the detection channel by an embedding method.
9. The vaccine detection apparatus according to claim 7, wherein the inner surface of the tube wall of the detection channel except for the reaction region, the detection region, and the control region is surface-hydrophobically treated.
10. A method for detecting a vaccine, which is performed using the vaccine detection apparatus according to any one of claims 1 to 6, the method comprising:
enabling the vaccine to be detected to enter the detection channel from the suction end of the detection channel through the driving assembly, and sequentially passing through the reaction region and the detection region in the driving direction;
when the inactivated or attenuated pathogen in the vaccine to be detected can be specifically combined with the first antibody marked by the colloidal gold in the reaction area and reacts with the first antibody in the detection area, so that the color of the detection area is displayed by the colloidal gold, the vaccine to be detected is an effective vaccine; when the inactivated or attenuated pathogen in the vaccine to be detected cannot react with the first antibody marked by the colloidal gold, the detection area does not display color, and the vaccine to be detected is an ineffective vaccine.
11. The method for detecting the vaccine according to claim 10, wherein a control area is further provided in the detection channel, the control area is located at one end of the detection area away from the reaction area, and the control area is provided with a second antibody, and the second antibody can be specifically combined with the first antibody;
the detection method of the vaccine further comprises the following steps: when the first antibody marked by the colloidal gold can react with the second antibody in the control area, so that the control area displays color through the colloidal gold, the detection is effective; and if the first antibody marked by the colloidal gold cannot react with the second antibody in the control area, the control area does not display color, and the detection is invalid.
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