A kit for implementing multiple connection detection and a preparation method thereof

Abstract

The present application relates to a kind of kit for realizing multiple detection and its preparation method.The preparation method of the kit for realizing multiple detection includes the following steps: nitrocellulose membrane is pasted on reagent strip back plate, and modification processing is formed in different detection projects chromatography flow channel, and coating is carried out in the detection point and quality control point in different detection projects chromatography flow channel;The corresponding binding pad of each project of multiple detection is cut into strip according to the space size of multiple detection reagent strip, and after array arrangement on binding pad substrate, it is pasted with interval, and multiple detection binding pad is obtained by cutting;Multiple detection binding pad, sample pad, absorbent paper and reagent strip back plate are positioned and pasted, and the kit for realizing multiple detection is obtained by cutting and installation.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological detection, and particularly relates to a kit for realizing multi-association detection and a preparation method thereof. BACKGROUND

[0002] Instant detection is a detection method that can be performed on site using portable analysis instruments and supporting reagents to quickly obtain detection results. Its advantages lie in the fact that it saves the complex sample pretreatment process, realizes immediate analysis on site and quickly obtains test results. The use of simple, user-friendly and portable test equipment promotes testing.

[0003] The lateral flow chromatographic reagent strip is a commonly used method in instant detection applications, and its principle is based on the adsorption and migration characteristics of chemicals on paper. The lateral flow chromatographic test strip has the advantages of simple operation, rapidness, portability, low cost and the like, and is therefore widely used in medical treatment, environmental protection, food safety and the like. For example, a urine lateral flow chromatographic test strip can be used to detect glucose, protein, ketone bodies and the like in urine, and is used to diagnose diabetes, kidney diseases and the like; a blood lateral flow chromatographic test strip can be used to detect cholesterol, blood sugar, hemoglobin and the like in blood, and is used to prevent cardiovascular diseases, diabetes and the like.

[0004] Generally, the lateral flow chromatographic test strip can only detect a single index of a single item, and multi-item detection can usually be realized only by using multiple reagent cards or installing multiple test strips in one reagent card. The test strips have different components, require multiple sample additions for separate detection, have higher sample addition requirements for users, are inconvenient to use, and the sample addition amount is difficult to control, and cannot be adapted to commonly used detection instruments for single item detection. A few reagent strips that can stably realize multi-association item synchronous detection have relatively harsh requirements for detection items, can only be applied to items with extremely similar detection environments, and still have mutual interference in the chromatographic process, which still affects the results, cannot be used for other item detection, are difficult to develop, and have poor universality.

[0005] Multi-association detection has the advantages of convenience and efficiency that single item detection reagent strips do not have, but most of them cannot realize mutual non-interference between multi-association item detection while adapting to commonly used detection instruments under the condition of considering preparation cost and operation convenience. Therefore, a preparation method of a multi-association detection reagent kit that has the advantages of mass production cost and use and good adaptability can truly exert the advantages of multi-association detection and meet market demand. SUMMARY

[0006] In view of the deficiencies of the prior art, the purpose of the present application is to provide a kit for realizing multi-association detection and a preparation method thereof.

[0007] In a first aspect, the present application provides a preparation method of a kit for realizing multiple detection, comprising the following steps:

[0008] pasting nitric acid fiber membrane on the back plate of the reagent strip and modifying and processing to form chromatographic flow channels of different detection items and coating detection points and quality control points in the chromatographic flow channels of different detection items;

[0009] cutting the binding pads corresponding to each item of the multiple detection into strips according to the space size of the multiple detection reagent strip and arraying and spacing pasting on the binding pad substrate, to obtain the multiple detection binding pad through cutting;

[0010] positioning and pasting the multiple detection binding pad, the sample pad, the water absorption paper and the reagent strip back plate, and obtaining the kit for realizing multiple detection through cutting and installation.

[0011] Preferably, the reagent strip back plate is provided with a mark hole for positioning near the edge of the shape.

[0012] Preferably, the modification and processing to form different chromatographic flow channels is UV glue curing.

[0013] Preferably, the different chromatographic flow channels are arranged in parallel with equal width, and the width of the single chromatographic flow channel is 2-2.5 mm.

[0014] Preferably, the width of the cutting of the binding pad of each detection item is not greater than the width of the chromatographic flow channel of the detection area of the corresponding item, preferably not greater than 2.5 mm, and more preferably 2 mm; and the length of the cutting of the binding pad of each detection item is not less than the length of the reagent strip back plate of the detection item.

[0015] Preferably, the binding pad substrate is a plastic sheet with a thickness of 0.1-0.2 mm, and the length of the binding pad substrate is not less than the whole page length of the multiple detection item test paper, preferably 320-340 mm.

[0016] Preferably, the binding pad substrate is a plastic sheet with a thickness of 0.1-0.2 mm, and the length of the binding pad substrate is not less than the whole page length of the multiple detection item test paper, preferably 320-340 mm.

[0017] Preferably, the binding pad substrate is provided with a void groove structure, and the void grooves are arranged in parallel and spaced on the binding pad substrate, and the adhesive pasting area of the binding pad is separated by the void groove structure, and the interval is preferably 3-5 mm, and more preferably the required length of the binding pad.

[0018] Preferably, the arrayed arrangement of the strip-shaped binding pad corresponding to each item and the binding pad substrate is in a vertical orthogonal manner; and the pasting interval of the strip-shaped binding pad of different detection items on the binding pad substrate is 0.7-1 mm, and preferably 1 mm.

[0019] In a second aspect, the present application provides a kit for realizing multiple detection obtained by the above preparation method, wherein the kit for realizing multiple detection comprises: a reagent strip back plate, and reagent strips corresponding to different detection items formed by detection area separation parts on the reagent strip back plate; preferably, each detection item reagent strip is sequentially and adjacently provided with a sample pad, a conjugate pad with a conjugate pad substrate, a nitrocellulose membrane and a water absorption paper on the reagent strip back plate in the direction of chromatographic liquid flow; more preferably, the sample pad is provided with a sample pad substrate.

[0020] Advantages

[0021] The present application solves the problem of mutual interference of detection items by combining the innovative method of conjugate pad combination and the separation of reaction areas, realizes reliable multiple detection in the smallest volume, and has a good effect on eliminating non-specific binding in detection. The innovative way of introducing the conjugate pad substrate of the conjugate pad in the multiple item detection is the core element of realizing mass production process of the multiple detection mode, which can reliably perform the separation processing of the conjugate under manual or equipment conditions, and is convenient for pasting and positioning on the final reagent plate, which is consistent with the existing production process, does not need additional production line arrangement and modification, and can realize mass production faster, has good operability, and excellent process stability.

[0022] In terms of cost, the multiple detection mode of the present application has excellent anti-interference ability, and integrates the reagent reaction in the smallest volume on a certain scale of reagent strip, so that the cost of the reagent strip and the cost of the supporting cartridge are greatly reduced compared with the form of multiple reagent strips and large reagent cartridges of the currently marketed multiple detection items, and the multiple detection mode has good mass production cost advantage.

[0023] The multiple detection mode of the present application is more friendly to users for use and operation, and only one time of sample addition is needed, which effectively solves the problem of sample addition amount of multiple sample additions. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 It is a schematic diagram of the reagent strip back plate pasted with nitrocellulose membrane;

[0025] Figure 2 It is a schematic diagram of the conjugate pad substrate provided with a mark hole as an example;

[0026] Figure 3 It is a schematic diagram of the conjugate pad and the conjugate pad substrate pasted in a vertical orthogonal manner for double detection as an example;

[0027] Figure 4 It is a schematic diagram of the double detection conjugate pad corresponding to the length of the conjugate pad after the conjugate pad substrate is bonded to the conjugate pad and then cut as an example;

[0028] Figure 5Fig. 1 shows the schematic diagram of the positioning and pasting of the cut multiple detection combined pad according to the mark hole and the reagent strip back plate;

[0029] Figure 6 Fig. 4 shows the schematic diagram of the pressing flow resistance structure of the blocking edge type cartridge at the sample window position;

[0030] Figure 7 Fig. 5 shows the comparison photo of the color development 5C of the reagent kit prepared in Example 1 and the reagent strip for single item detection;

[0031] Reference signs:

[0032] 1, nitrocellulose membrane, 2, reagent strip back plate, 3, back plate mark hole, 4, back plate shape, 5, detection area separation part,

[0033] 6, combined pad substrate, 7, combined pad substrate mark hole, 8, reagent paper whole plate length, 9, empty slot. DETAILED DESCRIPTION

[0034] The present application is further illustrated by the following embodiments, which should be understood as merely illustrative of the present application, but not limiting the present application.

[0035] Hereinafter, the preparation method of the reagent kit for multiple detection provided by the present application is exemplarily described. The preparation method can include the following steps.

[0036] (1) Back plate pasting nitrocellulose membrane (NC membrane). As shown in Figure 1 , the nitrocellulose membrane 1 is pasted on the reagent strip back plate 2 by back adhesive, and other isolation membranes on the reagent strip back plate 2 are not torn off. In some embodiments, the reagent strip back plate 2 is provided with a mark hole 3 for positioning near the edge of the back plate shape 4. The back plate mark hole 3 can be formed by secondary processing and punching after the reagent strip back plate 2 is formed, and the shape of the back plate mark hole 3 is preferably circular, and other geometric shape holes that can be used for positioning can also be used.

[0037] (2) Detection area division. The reagent strip back plate 2 pasted with the nitrocellulose membrane 1 in step (1) is positioned by using the back plate mark hole 3 and the back plate shape 4, and the detection area separation part 5 is modified and processed on the nitrocellulose membrane 1 according to the multiple detection requirements of the items to divide and form the chromatographic flow channels of different detection items, so that the detection sample liquid and the combined pad are not mixed after being combined.

[0038] In some embodiments, the modification of the detection area partition 5 can be performed by UV glue curing. It should be noted that the modification of the detection area partition 5 is not limited to this, as long as the nitrocellulose membrane 1 can be partitioned to form different detection areas, and the flow channel modification processing method can be used to prevent the detection sample liquid and the conjugate pad from flowing into each other after being combined.

[0039] When the modification is performed by UV glue curing, a UV light curing glue with hydrophobicity, biological safety, and stable physical and chemical properties after curing can be selected. After dispensing, a corresponding light source (preferably a UV laser curing lamp) is selected to cure and form the flow channel.

[0040] In some embodiments, the different chromatographic flow channels are preferably arranged in parallel with equal widths, that is, the detection area partition 5 is preferably a straight line to divide the different detection areas. The width of the single chromatographic flow channel can be set to 2-2.5 mm. The width of the detection area partition 5 is preferably 0.4-0.6 mm.

[0041] (3) Coating of detection points and quality control points. The detection points and quality control points corresponding to the multiple detection items are coated in the different detection flow channels of the nitrocellulose membrane 1 prepared in step (2), and the detection points and quality control points of the same item are controlled to be on the same side. The coating of the detection points and quality control points can be performed by using a dispensing device, and after dispensing, the membrane is placed in an oven for baking and curing.

[0042] In some embodiments, the reagent strip for multiple detection provided by the present application can also be coated by using the traditional reagent strip coating method, that is, the coating of the detection points and quality control points is performed before the modification of the detection area.

[0043] (4) Cutting of conjugate pad. The conjugate pad corresponding to each detection item is cut into a strip according to the size of the multiple detection reagent strip. The width of the conjugate pad for each detection item is controlled to be not greater than the width of the chromatographic flow channel of the detection area corresponding to the item, for example, the width of the conjugate pad can be controlled to be not greater than 2.5 mm, and is preferably 2 mm; the length of the conjugate pad for each detection item can be not less than the length of the reagent strip backboard of the detection item.

[0044] (5) Paste the conjugate pad on the conjugate pad substrate. The strip-shaped conjugate pad corresponding to each detection item cut in step (4) is arrayed and pasted on the conjugate pad substrate; then, the conjugate pad substrate pasted with the strip-shaped conjugate pad corresponding to each detection item is fixed by using a cutting instrument, and the multiple detection conjugate pad (conjugate pad combination strip) corresponding to the length of the conjugate pad of the multiple detection items is obtained by cutting.

[0045] In some embodiments, the binding pad substrate 6 can be a plastic sheet with a thickness of 0.1-0.2 mm, and the length of the binding pad substrate 6 is not less than the length of the whole test strip of the multi-test item; preferably, the length of the substrate can be between 320-340 mm, so that the effective length of the large plate formed after the process is completed is basically consistent with the conventional large plate length (300 mm).

[0046] As shown in Figure 2 In order to facilitate mass production, a binding pad substrate mark hole 7 corresponding to the multi-test reagent strip back plate 2 can be provided at the edge of the binding pad substrate 6 to position the relative position of the reagent binding pad substrate 6 and the reagent strip back plate 2, so that the multi-test binding pad (binding pad combination strip) can be accurately aligned with the detection area corresponding to each item of the multi-test reagent strip, and the whole test strip can be cut according to the fixed strip width to obtain a completely consistent multi-test reagent strip.

[0047] In some embodiments, the binding pad substrate 6 is also provided with a void slot structure 9. As shown in Figure 2 The void slots 9 are arranged in parallel on the binding pad substrate 6, and the adhesive area of the binding pad is separated by the void slot structure, and the interval is preferably 3-5 mm, and more preferably the length of the required binding pad. The length of the void slot 9 can be set to be greater than the length of the whole reagent strip (detection area), and the width can be set to be 0.4-0.6 times the length of the required binding pad or sample pad, and preferably 0.5 times. The design of the void slot structure 9 on the binding pad substrate 6 can ensure that the binding pad still has a spare section after cutting, so that the binding pad can be well connected to the detection NC membrane. If there is no void slot structure 9, the whole binding pad combination strip cannot be well pasted and connected to the NC membrane.

[0048] In some embodiments, the array arrangement of the strip-shaped binding pad corresponding to each item and the binding pad substrate can be a vertical orthogonal manner, so that the multi-test binding pad formed after cutting can correspond to each layer of chromatographic flow channel arranged in parallel; preferably, the paste interval of the strip-shaped binding pad of different detection items on the binding pad substrate can be 0.7-1 mm, and preferably 1 mm. As shown in Figure 3 The figure shows a schematic diagram of the binding pad and the binding pad substrate for double-test in a vertical orthogonal manner. Of course, the reagent strip provided by the present application is not limited to the case of double-test, but also can be the case of triple-test, quadruple-test and more. That is, the array arrangement of the strip-shaped binding pad corresponding to each detection item and the substrate is also applicable to the arrangement of the binding pad and the substrate for more test items.

[0049] As shown in Figure 4 The double detection bonding pad is cut after the bonding pad substrate is bonded with the bonding pad, and the corresponding bonding pad length of the double detection bonding pad is detected.

[0050] (6) Assembly. The multi-detection bonding pad cut in step (5) is positioned and pasted with the reagent strip back plate according to the mark hole 7, and the bonding pad of each detection item is connected with the corresponding detection area flow channel, as shown in Figure 5 At the same time, the sample pad and the water-absorbing paper are pasted on the reagent strip back plate, and the corresponding sample pad of each detection item is connected with the bonding pad, and the water-absorbing paper is connected with the detection area flow channel formed by the nitrocellulose membrane; then, the positioning hole is installed on the cutting tool for positioning and cutting according to the size of the multi-detection item, and the cutting width is the final required width of the multi-detection reagent strip; the cut multi-detection reagent strip is installed in the reagent cartridge to obtain the reagent kit for realizing multi-detection.

[0051] In some embodiments, especially for the preparation of multi-detection reagent kits that require different sample pads, the preparation of multi-detection sample pads can also be achieved by introducing a sample pad substrate. The cutting process of the sample pad corresponding to each detection item, the process of pasting the sample pad on the sample pad substrate and cutting, and the assembly process can refer to the above-mentioned process of pasting, cutting and assembling the bonding pad and the bonding pad substrate.

[0052] In addition, to match this type of reagent strip, preferably, the cartridge can be selected to have a pressure-blocking flow structure at the sample window position to prevent the sample liquid from directly spreading to the area of the bonding pad, thereby reducing the risk of cross-contamination of the bonding pad material between different detection items, and the blocking edge form is preferred, as shown in Figure 6

[0053] The preparation method provided by the application realizes the effect that the reactions in different detection areas do not interfere with each other in multi-detection by using UV ultraviolet curing glue to separate different detection areas. At the same time, by introducing the bonding pad substrate and the sample pad substrate, the bonding pads and sample pads for different item detection are arrayed and spacedly pasted on the substrate, and then cut once, which better realizes the processability of mass production of sample pads and bonding pads, has wide applicability, low cost and convenient operation.

[0054] ​The application solves the problem of mutual interference of detection items by combining the innovative method of the binding pad and the separation of the reaction area, and realizes reliable multi-connection detection in the smallest volume, which has a good effect on eliminating non-specific binding in detection. The innovative way of introducing the binding pad substrate of the binding pad in multi-item detection is the core element of realizing mass production process of this multi-connection detection mode. This way can reliably separate the binding under manual or equipment conditions, and is convenient for sticking and positioning on the final reagent plate, which is consistent with the existing production process, does not need additional production line arrangement and modification, and can realize mass production faster, has good operability and excellent process stability.

[0055] In terms of cost, the multi-connection detection mode of the application has excellent anti-interference ability, and integrates the smallest reagent reaction on a certain scale of reagent strip, so that the cost of the reagent strip and the cost of the matching cartridge are greatly reduced compared with the form of multi-reagent strip and large reagent cartridge of the multi-connection detection item on the market, and the cost advantage of mass production is good. At the same time, the multi-connection detection item of this mode is more friendly to users for use and operation, and only one time of adding sample is needed, which effectively solves the problem of adding sample quantity in multiple times.

[0056] The reagent kit for realizing multi-connection detection obtained by the preparation method provided by the application comprises: a reagent strip back plate, and reagent strips corresponding to different detection items formed by a detection area separation part; preferably, each detection item reagent strip is sequentially and adjacently provided with a sample pad, a binding pad with a binding pad substrate attached, a nitrocellulose membrane and a water absorption paper on the reagent strip back plate in the direction of chromatography liquid flow; more preferably, the sample pad is attached with a sample pad substrate.

[0057] The reagent kit for realizing multi-connection detection provided by the application ensures that it can still be well adapted to the commonly used single-item detection instrument in the case of reducing the external shape space. The reagent kit for realizing multi-connection detection items (such as double-connection detection and triple-connection detection) can be conveniently assembled by adjusting the size of each detection item reagent strip on the basis of the external shape of the existing single-connection detection cartridge, which is better than the current side-by-side or large-size multi-connection detection form.

[0058] In some embodiments, the length of the reagent kit for realizing multi-connection detection is 80-110 mm, the width is 19-25 mm, and the thickness is 5-7 mm.

[0059] The following examples further illustrate the present invention in detail. It should also be understood that the following examples are only for further explanation of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-essential improvements and adjustments made by those skilled in the art based on the above description of the present invention are within the scope of protection of the present invention. The specific process parameters, etc., in the following examples are merely examples within a suitable range; that is, those skilled in the art can make appropriate selections within the appropriate range based on the description herein, and are not intended to be limited to the specific values ​​in the examples below.

[0060] Example 1

[0061] The preparation method of the kit for detecting influenza A and B and novel coronavirus is as follows:

[0062] (1) Paste the blank NC film onto the backing plate. The backing plate has dimensions of 300mm×60mm×0.5mm.

[0063] (2) Place the backing plate with the nitrocellulose membrane onto a dedicated dispensing device, fix it with a tooling, and use UV-curable adhesive to modify the detection area, dividing the detection area into two parallel and non-interfering detection areas; the flow channel width of a single detection area is 2.2mm (0.6mm for the detection area division).

[0064] (3) Use a film dotting device to coat the quality control points and test points of the novel coronavirus detection project and the influenza A and B detection project on the NC membrane. The test points and quality control points of the same project are on the same side. After the film dotting is completed, place it in an oven for baking and curing.

[0065] (4) Process the conjugate pads for the novel coronavirus test and the conjugate pads for the influenza A and B test separately, and cut them into strips according to the specification of 300mm×2mm;

[0066] (5) Arrange the strip-shaped bonding pads cut for each item vertically and orthogonally on the bonding pad substrate and then stick them at intervals, with a 1mm gap between the bonding pads. The bonding pad substrate size is 320mm×300mm. Then, fix the bonding pad substrate with the strip-shaped bonding pads corresponding to each test item with a strip cutting instrument and cut a bonding pad combination strip with a width of 6mm and a length of 300mm.

[0067] (6) The cut combined pad assembly strip is pasted to the corresponding area of the reagent strip back plate, and the sample pad and the absorbent paper are pasted on the back plate, and then the positioning hole is installed on the special cutting tooling, the cutting position is adjusted, the separation part of the detection area on the reagent strip is ensured to be in the middle of the cut reagent strip, the reagent plate is cut into a 5mm wide reagent strip; the cut reagent strip is installed into the reagent cartridge, packaged, and the buffer, instruction manual, nasal swab and other accessories are placed in the packaging box to obtain the reagent kit for realizing the detection of influenza A (H1N1), influenza B (H3N2) and new coronavirus.

[0068] The reagent kit prepared in Example 1 is subjected to sample addition test, and the detection is compared with the reagent strip for separate detection. The sample addition comparison chroma result value is shown in Table 1 as follows:

[0069] New coronavirus quality control concentration (unit / mL) 10 pg 100 pg 400 pg 800 pg 1 ng 2 ng 10 ng Single joint detection reagent strip color (C) 2 3 4.5 5 7 8 10 Multi joint detection reagent strip color (C) 2 3 4.5 5 7 8 10 FluA quality control B61140 (dilution multiple) 1.5w 3 wx 6 wx 12 wx Single joint detection reagent strip color (C) 7 6 5 4 Multi joint detection reagent strip color (C) 7 6 5 4 FluB quality control B62122 (unit / mL) 10 ng 50 ng 100 ng Single joint detection reagent strip color (C) 5 6 7 Multi joint detection reagent strip color (C) 5 6 7 .

[0070] Figure 7 The contrast photo of the color development of the reagent kit prepared in Example 1 and the reagent strip for separate detection of 5C.

[0071] At present, the detection of influenza A (H1N1), influenza B (H3N2) and new coronavirus generally needs to be detected respectively or twice sample addition on two reagent strips, and the detection operation is more, the sample addition amount is difficult to control, and the overall cost is higher. The multi-connection detection reagent kit prepared by the preparation method provided by the present application can simultaneously detect new coronavirus and three items of influenza A (H1N1), influenza B (H3N2) on one reagent kit, realize the simplification of use and operation, eliminate the influence of sample addition amount, and more reduce the cost, shorten the detection time, and the result is simple and clear.

[0072] The technical principles of the present application are described above in combination with specific embodiments. These descriptions are only for explaining the principles of the present application, and cannot be explained as limiting the protection scope of the present application in any way. Based on the explanation here, other specific embodiments of the present application can be conceived by those skilled in the art without creative labor, and these embodiments will fall within the protection scope of the present application.

Claims

1. A method for preparing a kit for multiplex detection, characterized in that, The preparation method comprises the following steps: The nitrocellulose membrane is pasted on the reagent strip back plate, and a modification treatment is performed to form chromatographic flow channels of different detection items and to coat detection points and quality control points in the chromatographic flow channels of different detection items; the chromatographic flow channels of different detection items are arranged in parallel at equal widths. The multi-detection combined pad corresponding to each item is cut into a strip shape according to the space size of the multi-detection reagent strip, and is arrayed and pasted on the combined pad substrate with intervals; the multi-detection combined pad is obtained through cutting; the combined pad substrate is provided with a void groove structure; the void grooves are arranged in parallel and with intervals on the combined pad substrate, and the adhesive pasting area of the combined pad is separated through the void groove structure; the arrayed manner of the strip-shaped combined pad corresponding to each item and the combined pad substrate is a vertical orthogonal manner. The multi-detection combined pad, the sample pad, the water absorption paper and the reagent strip back plate are positioned and pasted, and are cut and installed to obtain the reagent kit for realizing multi-detection.

2. The production method according to claim 1, characterized by, The reagent strip back plate is provided with a mark hole near the edge of the outer shape for positioning.

3. The production method according to claim 1, characterized by, The modification treatment forms chromatographic flow channels of different detection items in the form of UV glue curing.

4. The method of claim 1, wherein, The width of a single chromatographic flow channel in the chromatographic flow channels of different detection items is 2-2.5 mm.

5. The preparation method according to claim 1, characterized in that, The width of the cut combined pad of each detection item is controlled to be not greater than the width of the chromatographic flow channel of the corresponding detection area, and the length of the cut combined pad of each detection item is controlled to be not less than the length of the detection item reagent strip back plate.

6. The production method according to claim 5, wherein The width of the cut combined pad of each detection item is controlled to be not greater than 2.5 mm.

7. The preparation method according to claim 5, characterized in that, The width of the cut combined pad of each detection item is controlled to be 2 mm.

8. The method of claim 1, wherein, The combined pad substrate is selected to be a plastic sheet with a thickness of 0.1-0.2 mm, and the length of the combined pad substrate is not less than the length of the multi-detection item test paper.

9. The production method according to claim 8, characterized by, The length of the combined pad substrate is 320-340 mm.

10. The method of claim 2, wherein, The combined pad substrate is provided with a combined pad substrate mark hole corresponding to the multi-detection reagent strip back plate at the edge.

11. The method of claim 1, wherein, The mutual interval of the void grooves is 3-5 mm.

12. The method of claim 11, wherein, The mutual interval of the void grooves is the required length of the combined pad.

13. The method of claim 1, wherein, The pasting interval of the strip-shaped combined pad of different detection items on the combined pad substrate is 0.7-1 mm.

14. The method of claim 13, wherein, The pasting interval of the strip-shaped combined pad of different detection items on the combined pad substrate is 1 mm.

15. A kit for multiplex detection prepared by the method of any one of claims 1-14. The reagent kit for realizing multi-detection comprises a reagent strip back plate and reagent strips corresponding to different detection items formed by a detection area separation part.

16. The kit for enabling multiple association testing according to claim 15, wherein, The reagent strips corresponding to each detection item are sequentially and adjacently provided with a sample pad, a combined pad pasted with a combined pad substrate, a nitrocellulose membrane and a water absorption paper on the reagent strip back plate in the direction of chromatographic liquid flow.

17. The kit of claim 16, wherein the kit is capable of performing multiplexed detection. The sample pad is pasted with a sample pad substrate.

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