CN111665354A - Shared sample injection microfluidic immunoassay joint inspection device - Google Patents
Shared sample injection microfluidic immunoassay joint inspection device Download PDFInfo
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Abstract
The invention provides a shared sample injection microfluidic immunoassay joint inspection device, which comprises a joint inspection strip, a washing supply unit and a waste liquid absorption and storage unit, wherein the washing supply unit is connected to a sample injection end of the joint inspection strip; the joint detection strip is concavely provided with a first micro-flow channel and a plurality of second micro-flow channels, the first micro-flow channel extends from the sample inlet end of the joint detection strip to the sample outlet end of the joint detection strip, and the plurality of second micro-flow channels are intersected and communicated with the first micro-flow channel; a sample storage tank is arranged at the inlet of the first microflow channel; a marker storage groove is arranged in each of the first microflow channel at intervals of the second microflow channels; item reaction detection grooves are arranged at one ends, far away from the first microfluidic channels, of the second microfluidic channels, and solid phase components used for capturing a plurality of targets or corresponding markers are fixedly arranged on the inner walls of the item reaction detection grooves respectively. The device has high detection sensitivity and avoids mutual interference of a plurality of items.
Description
Technical Field
The invention relates to a shared sample injection microfluidic immunoassay joint inspection device, and belongs to the technical field of immunoassay.
Background
In the medical diagnosis process, a plurality of indexes are required to be jointly detected for the majority of disease diagnosis, and mutual evidence is adopted to assist the disease diagnosis, so that missed diagnosis or misdiagnosis is avoided, and the diagnosis accuracy is improved. Meanwhile, the multi-index combined detection is also used for clinically differential diagnosis of diseases, such as in the investigation and diagnosis of new coronavirus (nCOVI) epidemic induced new coronary pneumonia (covi-19), and for diagnosis of a patient or a suspected population, the patient should be informed of whether the patient is infected by the new coronavirus (nCOVI) or not, and if not, the patient should be informed of the disease state caused by other pathogens as much as possible, so that the multi-index combined detection is also required. Therefore, there is an important practical need to develop a real-time diagnosis of multi-index joint detection.
At present, the multi-index joint detection mostly adopts the labeled immune joint detection based on lateral chromatography, and the method generally has two detection methods. The first is to arrange different detection items in sequence and solid-phase on the same nitrocellulose solid-phase membrane (NC membrane), label objects of different items are layered together, when in detection, a sample permeates into a label object storage position from top to bottom and is combined with a corresponding label antigen or antibody, enters the NC membrane through a contact area with the NC membrane, moves to a solid-phase area by virtue of capillary action and is combined and deposited with a solid-phase component for the second time to form a visible characteristic detection band, and the defect is that the different items possibly interfere with each other, especially the corresponding antigen and antibody of one molecule interfere with each other and cannot be detected simultaneously, and the number of the detection indexes is limited. The second method is to prepare different items into independent test strips, then place the independent test strips in different card slots of the same plastic card respectively, and share a sample adding pool or add samples independently respectively, but the detection efficiency is low.
Disclosure of Invention
The invention provides a shared sample injection microfluidic immunoassay joint inspection device which can effectively solve the problems.
The invention is realized by the following steps:
a shared sample injection microfluidic immunoassay joint detection device is used for detecting a plurality of targets and comprises a joint detection strip, a washing supply unit and a waste liquid absorption and storage unit, wherein the washing supply unit is connected to the sample injection end of the joint detection strip, and the waste liquid absorption and storage unit is connected to the sample outlet end of the joint detection strip;
the joint detection strip is concavely provided with a first micro-flow channel and a plurality of second micro-flow channels, the first micro-flow channel extends from the sample inlet end of the joint detection strip to the sample outlet end of the joint detection strip, and the plurality of second micro-flow channels are intersected and communicated with the first micro-flow channel; a sample storage tank is arranged on the first micro-flow channel and close to the sample introduction end of the joint detection strip; a plurality of second microfluidic channels are arranged in the first microfluidic channel, and a plurality of label storage tanks are arranged at intervals of the second microfluidic channels and are respectively filled with labels corresponding to a plurality of targets; a plurality of project reaction detection grooves are arranged at one end, far away from the first microfluidic channel, of each second microfluidic channel, the inner walls of the project reaction detection grooves are fixedly provided with solid phase components used for capturing a plurality of targets or corresponding markers respectively, and observation windows are arranged on the walls of the joint detection strips corresponding to the bottoms or the side surfaces of the project reaction detection grooves; a film covering seal is arranged on the upper surface of the joint detection strip, so that a sealed communicating cavity channel is formed by the first microfluidic channel, the plurality of second microfluidic channels, the sample storage tank, the marker storage tank and the item reaction detection tank; a membrane-breaking prick extends from the sample introduction end of the joint inspection strip to the washing supply unit, and a channel is arranged in the membrane-breaking prick and communicated with the first microfluidic channel;
the washing supply unit comprises a washing liquid sleeve, a washing liquid pipe and a first sealing ring; the washing liquid pipe is arranged in the washing liquid sleeve, the first sealing ring seals the inlet end of the washing liquid sleeve and is provided with a first hollow matched with the cross section of the sample inlet end of the joint detection strip in shape, the sample inlet end of the joint detection strip extends into the washing liquid sleeve from the first hollow, the pipe orifice of the washing liquid pipe is coated with a sealing film, and the film breaking pricks at the sample inlet end of the joint detection strip are opposite to the sealing film; the washing liquid pipe is filled with washing liquid; when the joint inspection strip advances to the washing supply unit, the sealing film is punctured by a membrane-breaking prick at the sample injection end of the joint inspection strip, and the cleaning solution in the liquid-washing tube flows into the first microflow channel in the joint inspection strip from the channel of the membrane-breaking prick for washing.
As a further improvement, the waste liquid sucking and storing unit comprises a waste liquid sleeve, a negative pressure forming valve plate and a second sealing ring; the negative pressure forming valve plate is arranged in the inner cavity of the waste liquid sleeve, the second sealing ring seals the inlet end of the waste liquid sleeve and is provided with a second hollow matched with the cross section shape of the sample outlet end of the joint detection strip, the center of the negative pressure forming valve plate is provided with a third hollow matched with the cross section shape of the sample outlet end of the joint detection strip, the sample outlet end of the joint detection strip extends into the waste liquid sleeve from the second hollow, the negative pressure forming valve plate is fixedly sleeved on the sample outlet end of the joint detection strip through the third hollow, and the first microflow channel is communicated with the inner cavity of the outlet end of the waste liquid sleeve; when the waste liquid absorbing and storing unit of the first microflow channel slides towards the sample outlet end far away from the joint detection strip, the negative pressure forming valve plate does not move, and negative pressure is generated in an inner cavity of the outlet end of the negative pressure forming valve plate and the waste liquid sleeve to suck the first microflow channel.
In a further refinement, the first microfluidic channel, the plurality of second microfluidic channels, the sample reservoir, the label reservoir, and the item reaction detection chamber are all surface treated to be hydrophilic.
As a further improvement, the marker is selected from colloidal gold, colored microspheres, fluorescent substances, lanthanide rare earth element Eu2+And quantum dots.
As a further improvement, the joint inspection strip is integrally formed by compression molding or injection molding.
As a further improvement, the sample reservoir has a larger capacity than the marker reservoir.
As a further improvement, a plurality of the second microfluidic channels are arranged in parallel and perpendicular to the first microfluidic channel.
As a further improvement, a connecting rod extends out of the outer wall of a waste liquid sleeve of the waste liquid absorbing and storing unit along the direction of the joint detection strip, a chute is arranged on the wall of a washing liquid sleeve of the washing and supplying unit, and the extending end of the connecting rod is inserted into the chute to clamp the waste liquid absorbing and storing unit and the washing and supplying unit together.
As a further improvement, the depth of the second microfluidic channel is greater than the depth of the first microfluidic channel.
The invention provides a using method of the shared sample injection microfluidic immunoassay joint inspection device, which comprises the following steps:
s1, adding a sample liquid into the sample storage tank for reaction;
s2, sliding the waste liquid absorbing and storing unit to the sample outlet end far away from the joint inspection strip at the speed of 2-5 mm/S;
s3, after the reaction is finished, the joint inspection strip is pushed to the washing supply unit until a falling feeling is felt, and at the moment, a membrane rupture prick pierces the sealing membrane of the liquid washing tube to wash;
and S4, observing or calculating the detection result through visual inspection, colorimetric or fluorescent counting on the observation window of the joint inspection strip after the washing is finished.
The invention has the beneficial effects that:
the invention relates to a shared sample introduction microfluidic immunoassay joint inspection device, wherein a first microfluidic channel and a plurality of second microfluidic channels are concavely arranged on a joint inspection strip, a sample storage tank and a marker storage tank are arranged in the first microfluidic channel, an item reaction detection tank is arranged in the second microfluidic channel, the inner walls of the item reaction detection tanks are respectively and fixedly provided with a solid phase component for capturing a plurality of targets and corresponding markers thereof, when a sample to be inspected is arranged in the sample storage tank, the sample liquid flows along with the first microfluidic channel through the capillary action, the targets in the sample liquid are combined with the corresponding markers through the marker storage tank, then the sample liquid enters the item reaction detection tank in the second microfluidic channel to be combined with the corresponding solid phase, and the results are observed, recorded, qualitatively or quantitatively reported through an observation window in a visual inspection, colorimetric, fluorescent and other modes, the solid phase and the marker are respectively arranged at different positions, so that the mutual interference of the detection of each target object is avoided.
The shared sample injection microfluidic immunoassay joint inspection device disclosed by the invention has the advantages that only one sample injection is needed in the sample storage tank, and the sample liquid can enter the plurality of marker storage tanks and the plurality of item reaction detection tanks through the capillary action to carry out the detection of a plurality of items, so that the detection speed is greatly improved.
The shared sample injection microfluidic immunoassay joint inspection device is provided with the washing supply unit, when reaction detection is finished, the joint inspection strip is pushed to the washing supply unit, the membrane breaking prick at the sample injection end of the joint inspection strip pierces the sealing membrane, cleaning liquid in the liquid washing pipe flows into the joint inspection strip from a channel of the membrane breaking prick for washing, and samples, markers and reaction residues remained in the shared sample injection microfluidic immunoassay joint inspection device are fully washed, so that the interference of the residues on the detection is avoided, and the detection sensitivity is remarkably improved.
The shared sample injection microfluidic immunoassay joint inspection device is also provided with a waste liquid absorption and storage unit, a negative pressure valve plate is arranged in the waste liquid absorption and storage unit, when the waste liquid absorption and storage unit slides to the sample outlet end far away from the joint inspection strip, the negative pressure forming valve plate does not move, negative pressure is generated in the inner cavity of the outlet end of the negative pressure forming valve plate and the waste liquid sleeve, the negative pressure can accelerate the flow of liquid in the first microflow channel and the second microflow channel, the reaction speed is improved, and the detection speed is further improved.
The shared sample injection microfluidic immunoassay joint inspection device is simple in structure and easy to operate, meets various application scenes, and reduces medical inspection cost.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
Fig. 1 is a schematic structural diagram of an entirety of a shared sample injection microfluidic immunoassay joint detection device provided in an embodiment of the present invention.
Fig. 2 is a schematic top view of a joint-detection strip of a shared-sample microfluidic immunoassay joint-detection device according to an embodiment of the present invention.
Fig. 3 is a schematic front view of a joint-detection strip of a shared-sample microfluidic immunoassay joint-detection device according to an embodiment of the present invention.
Reference numerals:
a joint test strip 1, a washing supply unit 2, a first micro-flow channel 11, a second micro-flow channel 12, a sample storage tank 13, a marker storage tank 14, an item reaction detection tank 15, a coating film 16 and a membrane breaking prick 17,
a washing supply unit 2, a washing liquid sleeve 21, a washing liquid pipe 22, a first sealing ring 23, a sealing film 24 and a sliding groove 25,
the waste liquid sucking and storing unit 3, the waste liquid sleeve 31, the negative pressure forming valve plate 32, the second sealing ring 33 and the connecting rod 34.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
In the description of the present invention, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
Referring to fig. 1 to 3, the present invention provides a shared sample injection microfluidic immunoassay joint detection device for detecting a plurality of targets, including a joint detection strip 1, a washing supply unit 2 and a waste liquid storage unit 3, wherein the washing supply unit 2 is connected to a sample injection end of the joint detection strip 1, and the waste liquid storage unit 3 is connected to a sample output end of the joint detection strip. Wherein, the one end that the sample liquid of joint inspection strip 1 got into is the introduction of sample end, and the other end that the sample liquid flows out is the appearance of going out end.
The concave first microflow channel 11 and a plurality of second microflow channel 12 of establishing on the joint test strip 1, first microflow channel 11 is followed the introduction port of joint test strip 1 extends to the appearance end of leading out of joint test strip 1, and is a plurality of second microflow channel 12 all with first microflow channel 11 is crossing and communicates.
The first microfluidic channel 11 is provided with a sample reservoir 13 near the sample inlet end of the joint test strip 1. A plurality of marker reservoirs 14 are disposed in the first microfluidic channel 11 at intervals of the second microfluidic channels 12, and a plurality of the marker reservoirs 14 contain a plurality of markers corresponding to the targets. An item reaction detection groove 15 is disposed at one end of each of the second microfluidic channels 12 away from the first microfluidic channel 11, and solid phase components for capturing a plurality of targets or corresponding labels are respectively and fixedly disposed on inner walls of the item reaction detection grooves 15. A plurality of the marker storage tanks 14 from the sample inlet end to the sample outlet end of the joint test strip 1 are respectively filled with a first marker, a second marker and an Nth marker corresponding to a first target, a second target and an Nth target; the solid phase components in the plurality of item reaction detection cells 15 from the sample introduction end to the sample discharge end of the joint test strip 1 are a first solid phase component, a second solid phase component, and an nth solid phase component (N is a natural number and is not less than 3) corresponding to a first target, a second target, and an nth target, respectively. In this embodiment, N ═ 7 is preferable. The marker and the corresponding solid phase component in the device are independently arranged at different positions, and the solid phase components are also independently arranged, so that the mutual interference of detection items is avoided, and the detection sensitivity is improved. The target may be a protein antigen or antibody, a hapten T3, T4, etc., and the solid phase is divided into an antibody, an antigen, an anti T3 antibody, an anti T4 antibody, etc., which may be an anti-protein antigen.
And observation windows are arranged on the walls of the joint inspection strips corresponding to the bottoms or the side surfaces of the plurality of item reaction detection grooves 15. When the observation is carried out by visual observation, the bottom can be seen from the right side, or the side wall can be seen from the side; when the colorimetric detection is used, the color is detected through the side wall; when the fluorescence is used for detection, the side wall and the bottom can be excited right above, and the side wall detects the fluorescence; the side walls and bottom can also be excited with the side walls and the top detects fluorescence. The results of visual and instrumental tests can be reported qualitatively or quantitatively.
Set up tectorial membrane 16 on the upper surface of joint inspection strip 1 and seal, make first miniflow channel 11, a plurality of second miniflow channel 12 sample storage tank 13 marker storage tank 14 reaches project reaction detects groove 15 and forms sealed intercommunication chamber way, does benefit to the formation of capillary action and follow-up waste liquid and inhales storage unit 3 formation negative pressure for the liquid flow rate improves the speed and the efficiency of detection.
The sample inlet end of the joint inspection strip 1 extends a membrane-breaking prick 17 towards the washing supply unit 2, and a channel is arranged in the membrane-breaking prick 17 and communicated with the first microflow channel 11.
The washing supply unit 2 includes a washing liquid sleeve 21, a washing liquid pipe 22 and a first sealing ring 23; the liquid washing tube 22 is arranged in the liquid washing sleeve 21, the first sealing ring 23 seals an inlet section of the liquid washing sleeve 21 and is provided with a first hollow matched with the cross section shape of the sample introduction end of the joint inspection strip 1, the sample introduction end of the joint inspection strip 1 extends into the liquid washing sleeve 21 from the first hollow, a tube opening of the liquid washing tube 22 is coated with a sealing film 24, and the film breaking pricks 17 at the sample introduction end of the joint inspection strip 1 are opposite to the sealing film 24; the washing liquid pipe 22 contains washing liquid. The lotion sleeve 21 is of a cylindrical structure integrated with injection molding, and the first sealing ring 23 is of a circular structure made of rubber or silica gel.
When the joint test strip 1 is pushed to the washing supply unit 2, the membrane-breaking prick 17 at the sample-feeding end of the joint test strip 1 pierces the sealing membrane 24, and the cleaning solution in the cleaning solution tube 22 flows into the first microfluidic channel 11 in the joint test strip 1 from the channel of the membrane-breaking prick 17 for washing.
The waste liquid sucking and storing unit 3 comprises a waste liquid sleeve 31, a negative pressure forming valve plate 32 and a second sealing ring 33. The end of the waste liquid casing 31 close to the joint inspection strip 1 is defined as an inlet end, and the other end is defined as an outlet end. The negative pressure forming valve plate 32 is arranged in the inner cavity of the waste liquid sleeve 31, the second sealing ring 33 seals the inlet end of the waste liquid sleeve 31 and is provided with a second hollow matched with the cross section shape of the sample outlet end of the joint detection strip 1, the center of the negative pressure forming valve plate 32 is provided with a third hollow matched with the cross section shape of the sample outlet end of the joint detection strip 1, the sample outlet end of the joint detection strip 1 extends into the waste liquid sleeve 31 from the second hollow, the negative pressure forming valve plate 32 is fixedly sleeved on the sample outlet end of the joint detection strip 1 through the third hollow, and the first microflow channel 11 is communicated with the inner cavity of the outlet end of the waste liquid sleeve 31; when the waste liquid sucking and storing unit 3 slides to the sample outlet end far away from the joint inspection strip 1, the negative pressure forming valve plate 32 does not move, negative pressure is generated in the inner cavity of the outlet end of the negative pressure forming valve plate 32 and the waste liquid sleeve 31 to suck the first microflow channel 11, so that the liquid flow in the first microflow channel 11, the plurality of second microflow channels 12, the sample storage tank 13, the marker storage tank 14 and the item reaction detection tank 15 is accelerated, and the reaction speed and the detection efficiency are improved.
The waste liquid sleeve 31 is of a cylindrical structure integrated with injection molding, and the second sealing ring 33 is of a circular structure made of rubber or silica gel.
The first microfluidic channel 11, the plurality of second microfluidic channels 12, the sample reservoir 13, the label reservoir 14, and the item reaction detection reservoir 15 are all surface-treated to have hydrophilicity, so as to facilitate capillary action to push liquid to flow.
The marker is selected from colloidal gold, colored microspheres, fluorescent substance and lanthanide rare earth element Eu2+And quantum dots. The device is suitable for detection by various detection means.
The joint inspection strip 1 is molded integrally by polymer or silicate glass through compression molding or injection molding, the first microfluidic channel 11, the plurality of second microfluidic channels 12, the sample storage tank 13, the marker storage tank 14 and the item reaction detection tank 15 are all grooves formed by the upper surface of the joint inspection strip 1 in a concave mode, and the grooves are covered with the coating film 16 to form sealed slit cavities which are hydrophilic, so that capillary action is formed to push liquid to flow.
The capacity of the sample reservoir 13 is larger than that of the marker reservoir 14, and preferably, the capacity of the sample reservoir 13 is 10 to 20 times larger than that of the marker reservoir 14. Within this range, a large amount of sample liquid can be added to the sample reservoir 13 at a time, and the sample liquid can fill all of the label reservoir 14 and the item reaction detection well 15, thereby allowing detection of a plurality of items by a single sample addition. If the factor is more than 20, the volume of the sample reservoir 13 becomes too large to cause capillary action, which is disadvantageous to the flow of liquid.
Preferably, a plurality of the second microfluidic channels 12 are arranged in parallel and perpendicular to the first microfluidic channel 11, the first microfluidic channel 11 passes through the middle point of the length of the plurality of the second microfluidic channels 12, and the item reaction detection grooves 15 can be arranged at both ends of the second microfluidic channels 12, so that the detection reaction can be observed from both sides of the joint detection strip, and the detection can be simultaneously performed by different detection means at both sides, thereby realizing more efficient detection.
A connecting rod 34 extends out of the outer wall of a waste liquid sleeve 31 of the waste liquid absorption and storage unit 3 along the direction of the joint inspection strip 1, a sliding groove 25 is formed in the wall of a washing liquid sleeve 21 of the washing and supply unit 2, and the extending end of the connecting rod 34 is inserted into the sliding groove 25 to realize clamping. Will through connecting rod 34 waste liquid is inhaled and is stored up unit 3, joint inspection strip 1 and washing supply unit 2 connects and fixes into a whole, in addition waste liquid sleeve pipe 31 with lotion sleeve pipe 21 is cylindricly, and whole sharing advances kind micro-fluidic immunoassay joint inspection device and forms a pen appearance structure like this, carries easily.
The length of the second microfluidic channel 12 is 10-15mm, the distance between the intervals of a plurality of the second microfluidic channels 12 is 10-15mm, the length of the first microfluidic channel 11 can be adjusted according to the number of the second microfluidic channels 12, the number of the second microfluidic channels 12 is preferably 3-10, and the length of the first microfluidic channel 11 is preferably 40-200 mm. The width of the first microfluidic channel 11 and the second microfluidic channel 12 is 1.5-2.5mm, preferably 2 mm. The depth of the second microfluidic channel 12 is greater than the depth of the first microfluidic channel 11, preferably, the depth of the second microfluidic channel 12 is 2.0-2.5mm, and the depth of the first microfluidic channel 11 is 1.0-1.5 mm. Under the sizes of the lengths and the depths, a narrow cavity is formed, so that the capillary action is facilitated, the liquid can be fully distributed, the flowing of the liquid is accelerated, and the detection efficiency is improved. The depth of the second microfluidic channel 12 is greater than that of the first microfluidic channel 11, so that the reacted target substance and the label can stay in the second microfluidic channel 12 for a longer time, fully react with the corresponding solid phase component, fully capture the solid phase component, and improve the detection accuracy and sensitivity.
The outlet end of the marker reservoir 14 is lower than the inlet end to facilitate the flow of fluid into the marker reservoir 14.
The sample storage tank 13 and the marker storage tank 14 are both oval, the oval shape has fewer water caltrops than the square shape, liquid flows more smoothly, no dead angle is left in washing, and the detection sensitivity is further improved.
The prepared shared sample injection microfluidic immunoassay joint inspection device is sealed and stored in a sealing bag.
The use method of the shared sample injection microfluidic immunoassay joint inspection device comprises the following steps:
opening the sealed bag of the shared sample microfluidic immunoassay joint detection device, adding a sample liquid, such as blood, into the sample storage tank 11, allowing the sample liquid to flow along the first microfluidic channel 11 to the label storage tank 14 under capillary action, allowing a first target in the sample liquid to combine with a first label in the label storage tank 14 to form a first reaction chain mixture, allowing the first reaction chain mixture to continuously flow in the first microfluidic channel 11 and the second microfluidic channel 12 under capillary action, allowing the first reaction chain mixture to flow into the item reaction detection tank, sufficiently reacting with a solid phase component corresponding to the first target, and fixing the first reaction chain mixture on the solid phase component. This is done until the second to Nth targets complete the detection reaction.
In the reaction process, the waste liquid sucking and storing unit 3 slides towards the sample outlet end far away from the joint detection strip 1 at a speed of 2-5mm/s, the negative pressure forming valve plate 32 is fixed on the sample outlet end of the joint detection strip 1 and does not move, negative pressure is generated in the inner cavity of the outlet end of the negative pressure forming valve plate 32 and the waste liquid sleeve 31, and liquid in the first micro-flow channel 11, the plurality of second micro-flow channels 12, the sample storage tank 13, the marker storage tank 14 and the item reaction detection tank 15 is sucked and enters the inner cavity of the waste liquid sleeve 31, so that the flow of the liquid is accelerated, and the reaction and detection speed is increased. The sliding speed is very critical, and if the sliding speed is less than 2mm/s, the formed negative pressure is too small, so that the reaction speed is difficult to accelerate; if the sliding speed is more than 5mm/s, the formed negative pressure is too large, the suction is too fast, the reaction of the target, the marker and the solid phase component is insufficient, and the detection sensitivity is reduced.
After the reaction is completed, the joint test strip 1 is held and pushed to the washing supply unit 2 until a falling feeling is sensed, at this time, the membrane rupture 17 ruptures the sealing membrane 24 of the liquid washing tube 22, and the washing liquid in the liquid washing tube 22 flows into the first microfluidic channel 11 in the joint test strip 1 by capillary action from the passage of the membrane rupture 17 to wash, and the first microfluidic channel 11, the plurality of second microfluidic channels 12, the sample storage tank 13, the marker storage tank 14, and the item reaction detection tank 15 are washed in sequence. The remaining sample solution, the labeling substance, etc. are washed away, and the reaction chain mixture immobilized on the solid phase component is not washed away. In the washing process, the waste liquid sucking and storing unit 3 can slide towards the sample outlet end far away from the joint inspection strip 1 at the speed of 6-10mm/s, and the cleaning liquid and the residual liquid can be sucked into the inner cavity of the waste liquid sleeve 31, so that the washing process is accelerated. This sliding speed is very critical, and if it is less than 6mm/s, it is difficult to clean, and if it is more than 10mm/s, the suction force becomes too large, and the solid phase component is easily extracted, which affects the detection sensitivity.
After washing is finished, the detection result can be observed and calculated through visual inspection, colorimetric or fluorescent counting on the observation window of the joint inspection strip 1, so that interference of residue on detection is avoided, and the detection sensitivity is greatly improved. When the visual observation is carried out, the bottom can be seen from the right side, or the side wall can be seen from the side; when the colorimetric detection is used, the color is detected through the side wall; when the fluorescence is used for detection, the side wall and the bottom can be excited right above, and the side wall detects the fluorescence; the side walls and bottom can also be excited with the side walls and the top detects fluorescence. The results of visual and instrumental tests can be reported qualitatively or quantitatively.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A shared sample injection microfluidic immunoassay joint inspection device is used for detecting a plurality of targets and is characterized by comprising a joint inspection strip, a washing supply unit and a waste liquid absorption and storage unit, wherein the washing supply unit is connected to the sample injection end of the joint inspection strip, and the waste liquid absorption and storage unit is connected to the sample outlet end of the joint inspection strip;
the joint detection strip is concavely provided with a first micro-flow channel and a plurality of second micro-flow channels, the first micro-flow channel extends from the sample inlet end of the joint detection strip to the sample outlet end of the joint detection strip, and the plurality of second micro-flow channels are intersected and communicated with the first micro-flow channel; a sample storage tank is arranged on the first micro-flow channel and close to the sample introduction end of the joint detection strip; a plurality of second microfluidic channels are arranged in the first microfluidic channel, and a plurality of label storage tanks are arranged at intervals of the second microfluidic channels and are respectively filled with labels corresponding to a plurality of targets; item reaction detection grooves are arranged at one ends, far away from the first microfluidic channels, of the second microfluidic channels, solid-phase components used for capturing a plurality of targets or corresponding markers are fixedly arranged on the inner walls of the item reaction detection grooves respectively, and observation windows are arranged on the walls, corresponding to the bottoms or the side faces of the item reaction detection grooves, of the joint detection strips; a film covering seal is arranged on the upper surface of the joint detection strip, so that a sealed communicating cavity channel is formed by the first microfluidic channel, the plurality of second microfluidic channels, the sample storage tank, the marker storage tank and the item reaction detection tank; a membrane-breaking prick extends from the sample introduction end of the joint inspection strip to the washing supply unit, and a channel is arranged in the membrane-breaking prick and communicated with the first microfluidic channel;
the washing supply unit comprises a washing liquid sleeve, a washing liquid pipe and a first sealing ring; the washing liquid pipe is arranged in the washing liquid sleeve, the first sealing ring seals the inlet end of the washing liquid sleeve and is provided with a first hollow matched with the cross section of the sample inlet end of the joint detection strip in shape, the sample inlet end of the joint detection strip extends into the washing liquid sleeve from the first hollow, the pipe orifice of the washing liquid pipe is coated with a sealing film, and the film breaking pricks at the sample inlet end of the joint detection strip are opposite to the sealing film; the washing liquid pipe is filled with washing liquid; when the joint inspection strip advances to the washing supply unit, the sealing film is punctured by a membrane-breaking prick at the sample injection end of the joint inspection strip, and the cleaning solution in the liquid-washing tube flows into the first microflow channel in the joint inspection strip from the channel of the membrane-breaking prick for washing.
2. The shared sample introduction microfluidic immunoassay joint inspection device of claim 1, wherein the waste liquid absorption and storage unit comprises a waste liquid sleeve, a negative pressure forming valve plate and a second sealing ring; the negative pressure forming valve plate is arranged in the inner cavity of the waste liquid sleeve, the second sealing ring seals the inlet end of the waste liquid sleeve and is provided with a second hollow matched with the cross section shape of the sample outlet end of the joint detection strip, the center of the negative pressure forming valve plate is provided with a third hollow matched with the cross section shape of the sample outlet end of the joint detection strip, the sample outlet end of the joint detection strip extends into the waste liquid sleeve from the second hollow, the negative pressure forming valve plate is fixedly sleeved on the sample outlet end of the joint detection strip through the third hollow, and the first microflow channel is communicated with the inner cavity of the outlet end of the waste liquid sleeve; when the waste liquid absorbing and storing unit slides to the sample outlet end far away from the joint inspection strip, the negative pressure forming valve plate does not move, and negative pressure is generated in an inner cavity of the outlet end of the negative pressure forming valve plate and the waste liquid sleeve to suck the first microflow channel.
3. The shared sample microfluidic immunoassay joint inspection device of claim 2, wherein the first microfluidic channel, the plurality of second microfluidic channels, the sample reservoir, the label reservoir and the item reaction detection chamber are all surface treated to have hydrophilicity.
4. The shared sample feeding microfluidic immunoassay joint detection device of claim 2, wherein the label is selected from colloidal gold, colored microspheres, fluorescent substance, lanthanide rare earth element Eu2+And quantum dots.
5. The shared sample introduction microfluidic immunoassay joint detection device of claim 2, wherein the joint detection strip is integrally molded by compression molding or injection molding.
6. The shared sample microfluidic immunoassay joint detection device of claim 2, wherein the sample reservoir has a larger capacity than the label reservoir.
7. The shared sample microfluidic immunoassay joint detection device of claim 2, wherein a plurality of the second microfluidic channels are arranged in parallel and perpendicular to the first microfluidic channels.
8. The shared sample introduction microfluidic immunoassay joint inspection device of claim 2, wherein a connecting rod extends out of the outer wall of the waste liquid casing of the waste liquid suction and storage unit along the direction of the joint inspection strip, a sliding groove is arranged on the wall of the washing liquid casing of the washing and supply unit, and the extending end of the connecting rod is inserted into the sliding groove to clamp the waste liquid suction and storage unit and the washing and supply unit together.
9. The shared-sample microfluidic immunoassay joint detection device of claim 1, wherein the depth of the second microfluidic channel is greater than the depth of the first microfluidic channel.
10. A method of using the shared-sample microfluidic immunoassay joint detection device of any of claims 1 to 9, comprising the steps of:
s1, adding a sample liquid into the sample storage tank for reaction;
s2, sliding the waste liquid absorbing and storing unit to the sample outlet end far away from the joint inspection strip at the speed of 2-5 mm/S;
s3, after the reaction is finished, the joint inspection strip is pushed to the washing supply unit until a falling feeling is felt, and at the moment, a membrane rupture prick pierces the sealing membrane of the liquid washing tube to wash;
and S4, observing or calculating the detection result through visual inspection, colorimetric or fluorescent counting on the observation window of the joint inspection strip after the washing is finished.
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