A detection system and detection method

Abstract

This invention provides a detection system and method using immunochromatography. The detection system includes a laterally flowing test strip and an incubation tube, the incubation tube containing a marker that specifically binds to the analyte. The detection system and method of this invention can improve product sensitivity and operating range, and can achieve multiplex detection.

Description

Technical Field

[0001] This invention belongs to the field of biological detection technology, and specifically relates to the structure of test strips, the preparation method of test strips, and detection methods. Background Technology

[0002] Severe Acute Respiratory Syndrome Coronavirus type 2 (SARS-CoV-2) is an enveloped, non-fragment-positive RNA virus. Studies have found that the RBD of the SARS-CoV-2 S protein strongly interacts with the human ACE2 receptor, leading to endocytosis into deep lung host cells and viral replication. Infection with SARS-CoV-2 triggers an immune response, resulting in the production of neutralizing antibodies in the blood. Neutralizing antibodies are protective antibodies produced by the immune system that recognize and prevent pathogens from binding to host cells, exerting a protective effect. For COVID-19, neutralizing antibodies competitively bind to the viral surface S protein, blocking the S protein and preventing viral invasion of cells. Neutralizing antibodies, as antiviral antibodies, constitute only a small fraction of all antiviral antibodies. Vaccination is widely recognized as the most economical and effective method for controlling viral outbreaks. To combat the COVID-19 pandemic, countries worldwide are accelerating the development of COVID-19 vaccines that induce neutralizing antibodies in recipients to protect against the virus.

[0003] Accurate and rapid evaluation of vaccination efficacy has always been a key focus of scientific research. Traditional methods for detecting neutralizing antibodies in viral vaccines, such as the PNRT, CPE, and pseudovirus neutralization assay, are the most commonly used methods. While these traditional methods are reliable, they are time-consuming, require strict standardization, and are inefficient, typically taking 2-4 days to complete. This poses a significant challenge for large-scale population-based evaluation of vaccination efficacy. Therefore, a simple and rapid alternative method is urgently needed for assessing protective antibodies in large populations. Based on this, a miniaturized, portable, easy-to-operate, low-cost method for immediate detection of neutralizing antibodies is urgently required.

[0004] Lateral flow test strips are commonly used point-of-care testing products that utilize chromatography to transfer samples across the strip and obtain test results. A typical lateral flow test strip includes a support plate, on which, stacked sequentially from upstream to downstream, are a sample application pad, a labeled conjugate release pad (or simply labeled pad), a detection pad, and an absorbent pad. The labeled pad contains a marker that binds to the analyte, such as latex, colloidal gold, or fluorescent microspheres labeled with antigens or antibodies. The detection pad usually has a test line and a reference line. Depending on the reaction principle, as the sample flows across the strip, the marker may or may not aggregate on the test line. The presence or concentration of the analyte is determined based on the marker signal, such as color or fluorescence. The reference line is used to determine the effectiveness of the test strip and for instrument positioning when reading test results. Lateral flow test strips often suffer from low sensitivity; improving sensitivity to better utilize them for evaluating neutralizing antibodies or other analytes is a pressing technical challenge. Summary of the Invention

[0005] To address the aforementioned problems of the prior art, the present invention provides a detection system comprising a lateral flow test strip and an incubation tube, wherein the incubation tube contains a marker that specifically binds to the analyte.

[0006] Furthermore, the test strip is mounted in the test plate.

[0007] Furthermore, the incubation tube and the test plate are two independently configured devices in the detection system.

[0008] Furthermore, the incubation tubes are mounted on the test plate.

[0009] Furthermore, the test strip consists of, from upstream to downstream, stacked sample pads, detection pads, and absorption pads.

[0010] Furthermore, the test strip also includes a flow guide pad located between the sample application pad and the detection pad.

[0011] Furthermore, the test strip does not contain any markers that can specifically bind to the analyte.

[0012] The present invention also provides a detection method for detecting an analyte, using the detection system described in the present invention; the detection steps include: adding the sample to an incubation tube to form a reconstitute of the sample and the marker, and incubating; adding the incubated reconstitute to a test strip; and obtaining the detection result based on the detection line on the test strip.

[0013] Furthermore, when adding the sample to the incubation tube, add the buffer solution to the incubation tube, or when adding the reconstituted solution to the test strip, add the buffer solution.

[0014] Furthermore, the analyte can be an antigen, antibody, or DNA / RNA nucleic acid aptamer.

[0015] The present invention also provides a method for preparing a detection system, including preparing a test strip and an incubation tube with a marker.

[0016] Furthermore, the markers are stored in the incubation tube in the form of dry reagents, liquid reagents, powdered reagents, or gel reagents.

[0017] Furthermore, the markers are prepared in incubation tubes by drying or lyophilization.

[0018] Furthermore, the labeled particles on the markers are selected from latex, colloidal gold, fluorescein, quantum dots, and upconversion fluorescent nanoparticles.

[0019] Furthermore, the test strip consists of, from upstream to downstream, stacked sample pads, detection pads, and absorption pads.

[0020] Furthermore, the test strip also includes a flow guide pad located between the sample application pad and the detection pad.

[0021] Furthermore, the test strip does not contain any markers.

[0022] Furthermore, the test strip is mounted in the test plate.

[0023] Furthermore, the incubation tube and the test plate are two independently configured devices in the detection system.

[0024] Furthermore, the incubation tubes are mounted on the test plate.

[0025] This invention transfers the marker from the labeling pad of the test strip to an incubation tube outside the test strip, allowing it to incubate together with the sample outside the test strip. This operation significantly improves the sensitivity and accuracy of the product. Compared with existing test strips with a labeling pad, it reduces the requirements for the test strip and simplifies the assembly process. It enables multiple detection, which not only facilitates operation but also reduces cross-influence caused by mixing different complexes through the labeling pad, thus minimizing the impact on detection accuracy. Attached Figure Description

[0026] Figure 1 In the detection system of this invention, the test plate and the incubation tube are connected and set up independently.

[0027] Figure 2 An exploded view of the test board described in this invention.

[0028] Figure 3 A schematic diagram of the test paper without a flow guide pad according to the present invention.

[0029] Figure 3-1 A schematic diagram of the test strip with multiple detection lines described in this invention.

[0030] Figure 3-2 This invention only includes a schematic diagram of the test strip for the test line.

[0031] Figure 4 A schematic diagram of the test paper with a flow guide pad according to the present invention.

[0032] Figure 5 The operational steps for sample analysis using the detection system of the present invention.

[0033] Figure 5-1 Another operational step for sample analysis using the detection system of the present invention.

[0034] Figure 6 The detection system of the present invention comprises a test plate and an incubation tube assembled together.

[0035] Figure 7 yes Figure 6 A diagram from another angle.

[0036] Figure 8 A schematic diagram showing the incubation tubes not mounted on the test plate.

[0037] Figure 9 A schematic diagram of an incubation tube at one angle.

[0038] Figure 10 Schematic diagram of the upper opening of the incubation tube.

[0039] Figure 11 A schematic diagram of the incubation tube after rotation.

[0040] Figure 12 A schematic diagram comparing the test results of the test panel with the color chart. Detailed Implementation

[0041] like Figure 1 The detection system using immunochromatography shown includes a test plate 100 and an incubation tube 200 separate from the test plate. The test plate includes a housing and test strips contained within the housing. Figure 2 , Figure 3 and Figure 3-1As shown, the housing includes an upper cover 111 and a lower plate 112, which are interlocked to hold the test strip 120 within the housing. The upper cover of the housing has a sample application hole 101 and an observation window 102. The test strips 120 and 120a are lateral flow test strips, which, from upstream to downstream, sequentially include a sample application pad 121, a detection pad 122, and an absorption pad 123 stacked on top of each other. The stacking means that one end of the sample application pad partially overlaps with one end of the detection pad; and one end of the absorption pad partially overlaps with the other end of the detection pad. In some preferred embodiments, the test strip also includes a base card 124, on which the sample application pad, detection pad, and absorption pad are adhered to increase the rigidity of the test strip. Figure 4 The test strip 120b shown is similar to... Figure 3 The test strips shown are basically the same, except that a flow guide pad 125 is provided between the sample pad and the test pad. In one embodiment, the flow guide pad does not contain reagents, i.e., it is a blank pad.

[0042] The detection pad 122 has detection lines 301, which include reagents that specifically bind to the analyte or to a marker. Depending on the detection requirements, the detection pad may include multiple detection lines, such as... Figure 3-1 The test strip 120a shown includes test lines 301a, 301b, 301c and 301d.

[0043] Depending on the testing requirements, the test pad may also include a reference line 302. The function of the reference line includes, but is not limited to, determining whether the test strip is effective; comparing the color of the test line with the color of the reference line to determine the presence of the analyte, or positioning the instrument when reading the test results. The test strip's measurement result on the sample can be obtained from the signals given by the test line and / or the reference line, for example, but not limited to, the following methods: comparing the test line and / or the reference line with the matching color chart; comparing the test line and the reference line visually or with an instrument; judging based on the change in the color of the test line before and after contact with the sample; comparing the test line visually or with an instrument on the blank area of ​​the test pad, where the blank area refers to the area on the test pad that is neither the test line nor the reference line.

[0044] The test strips 120, 120a, and 120b do not include a marker that specifically binds to the analyte; the marker is placed inside the incubation tube 200. The marker can be pre-stored in the incubation tube or placed in by the operator during testing. The marker in the incubation tube can be stored in various forms, such as dry reagent, liquid reagent, powder reagent, or gel reagent. The marker can also be pre-treated on a carrier before being placed in the incubation tube; for example, a reagent containing the marker can be sprayed onto glass fiber, dried, and then a glass fiber containing a certain amount of the marker can be placed in the incubation tube. When the incubation tube pre-stores the marker reagent, preferably, the incubation tube also includes a sealing element, such as aluminum foil, a plastic cap, or a plastic seal. Figure 1 As shown, the incubation tube 200 includes a tube body 211, a marker 303 pre-loaded inside the tube, a closed bottom, and a sealing element 212, such as aluminum foil, covering the opening of the incubation tube. In some embodiments, the incubation tube may also be referred to as a reaction tube.

[0045] like Figure 5 and Figure 5-1 In the detection procedure shown, the seal of the incubation tube is removed, and the sample is first mixed with the marker inside the incubation tube to form a reconstitute. Then, the reconstitute is added to the sample application pad of the test strip. Depending on the reaction principle, as the sample flows on the test strip, the marker may or may not aggregate on the detection line. The presence or concentration of the analyte is determined based on the signal from the marker on the detection line, such as a color signal or a fluorescence signal.

[0046] The sample application pad is made of an absorbent material, such as glass fiber, polyester film, non-woven fabric, polyethersulfone, or polysulfone, with a thin film material being preferred. The test pad is made of materials such as nitrocellulose, glass fiber, polyethersulfone, and nylon, with nitrocellulose film being preferred. The absorbent pad is made of an absorbent material, such as filter paper. The base card can be made of a hydrophobic material such as polyvinyl chloride to ensure that the sample does not leak from the bottom support. The incubation tube is made of materials suitable for storing reagents, such as ethylene plastic or glass.

[0047] like Figures 6 to 11 The detection system shown includes a test plate 100a and an incubation tube 400 mounted on the test plate. Figure 6 and 7 The test board shown is Figure 1 The test panels in the examples are basically the same, each containing test strips. Test panel 100a and incubation tube 400 can be molded as a single piece, or they can be two separate components assembled together through subsequent processing. Figure 8 As shown, an incubation tube mounting base 103 is provided at the sample application port 101a of the test plate. (As indicated...) Figure 9 and 10 As shown, the incubation tube is a tube with openings 401 at both the top and bottom. The bottom of the incubation tube is installed on the sample dispensing base. If the incubation tube 400 contains a pre-stored marker 303, the top opening and / or bottom opening of the incubation tube can be sealed with a sealant. The sealant is removed when the sample is added to the incubation tube.

[0048] During sample incubation in the incubation tube, or before the mixture of marker and sample is added to the sample pad through the sample application well of the test plate, the bottom opening of the incubation tube on the test plate is not in communication with the liquid in the sample application well of the test plate. Only after incubation is complete and the incubated mixture is added to the sample pad does the bottom opening of the incubation tube communicate with the liquid in the sample application well of the test plate. The mixture flows through the sample pad to the detection pad to complete the detection. In one design, the bottom opening of the incubation tube is sealed with a sealant such as aluminum foil. After the incubation tube is mounted on the test plate, the bottom opening of the incubation tube is aligned with the sample application well of the test plate, and the liquid communication between the two is blocked by the aluminum foil sealant. After incubation is complete, the aluminum foil sealant is punctured with a pointed tool, allowing the liquid in the incubation tube to flow into the test plate. In another design, an O-ring is installed around the lower outlet of the incubation tube. After the incubation tube is mounted on the test plate, the bottom opening of the incubation tube is not aligned with the sample dispensing hole of the test plate, but rather offset from it. The O-ring blocks the liquid flow between the outlet of the incubation tube and the sample dispensing hole. When the incubation tube is rotated, the bottom opening of the incubation tube aligns with the sample dispensing hole of the test plate, releasing the liquid, which enters the test plate, and reaches the sample pad. Figure 6 and Figure 11 As shown, after mounting the incubation tube on the test plate, the bottom opening of the incubation tube is not aligned with the sample well of the test plate, but rather offset from it, preventing liquid flow between the incubation tube and the sample well. After incubation is complete, the incubation tube is rotated 400 degrees to open from the sample well. Figure 6 The incubation location was changed to Figure 11 The detection position is shown in the figure. At this time, the bottom opening of the incubation tube is aligned with the sample application hole of the test plate, and the liquid in the incubation tube flows into the test plate.

[0049] In addition to the forms described above, incubation tubes can also be centrifuge tubes, with the caps that come with the centrifuge tubes serving as seals for the tube openings.

[0050] In other embodiments, the incubation tubes containing markers that specifically bind to the analyte are not sealed with seals. For example, the incubation tubes containing the markers without seals are packaged together with test strips or test plates in an aluminum foil bag for later use.

[0051] The analyte can be an antigen, antibody, or DNA / RNA aptamer. The labeling particles (labeling reagents) used to label the antigen, antibody, or DNA / RNA aptamer can be selected from latex, colloidal gold, fluorescein (fluorescent microspheres), quantum dots, upconversion fluorescent nanoparticles, etc.

[0052] The labeled substance is a complex, formed by the combination of a substance that specifically binds to the analyte and a labeled particle with an indicative signal. For example, a labeled substance is a complex formed by labeling specific antigens, antibodies, or DNA / RNA aptamers with the aforementioned labeling reagents such as latex, colloidal gold, fluorescein (fluorescent microspheres), quantum dots, or upconversion fluorescent nanoparticles. For instance, a complex of colloidal gold and an antigen is a labeled substance, often simply called a gold label.

[0053] The clinical samples used for testing by the device and method described in this invention can be blood (serum, plasma or whole blood), urine, cerebrospinal fluid or other clinical samples, or feces or other clinical samples that need to be pre-treated.

[0054] The detection system of the present invention can also employ a combination of test strips and incubation tubes, wherein the test strips are not installed within the test plate. The test strip is a lateral flow test strip, comprising, from upstream to downstream, stacked sample application pad 121, detection pad 122, and absorption pad 123. The test strip also includes a base card 124, on which the sample application pad, detection pad, and absorption pad are adhered. The test strip may also include adhesive stickers covering the upper surfaces of the sample application pad and the absorption pad, respectively. In another embodiment, the test strip may further include a flow guide pad 125 disposed between the sample application pad and the detection pad. In one embodiment, the flow guide pad is empty of reagents, i.e., a blank pad. After the sample has incubated with the marker in the incubation tube, the test strip is placed directly into the incubation tube, allowing the sample application pad to contact the incubated sample, or the incubated sample in the incubation tube is dripped onto the sample application pad of the test strip. The incubated sample flows through the sample application pad into the detection pad for detection, and any excess sample on the test strip is absorbed by the absorption pad.

[0055] Example 1: Detection system and method for SARS-CoV-2 neutralizing antibodies in samples.

[0056] I. Detection Reaction Principle:

[0057] The test strip has two lines coated on the test pad: a test line coated with ACE-2 antigen and a control line (also known as a reference line). The test pad is made of an NC membrane. The incubation tube contains a dried gold label, a complex of colloidal gold and the SARS-CoV-2 S recombinant antigen. When there are no neutralizing antibodies in the sample, the gold-labeled complex of colloidal gold and S recombinant antigen migrates laterally along the NC membrane under capillary action. When this gold-labeled complex passes the test line, due to the high affinity of the S antigen for ACE-2, it is captured by the ACE-2 immobilized on the NC membrane, and the test line shows color. When there are neutralizing antibodies in the sample, the neutralizing antibodies in the sample first come into contact with the gold-labeled complex, and the S antigen first binds to the neutralizing antibodies, thus it does not bind to the ACE-2 immobilized on the NC membrane, and the test line does not show color. Regardless of the presence of neutralizing antibodies in the sample, the reference line will show color.

[0058] II. Preparation of the Detection System

[0059] Step 1. Adhere the NC membrane, which serves as the detection pad, to the substrate. Use a dotting machine to coat the NC membrane with ACE-2 antigen diluted to a certain concentration to form a detection line. The C line is composed of secondary antibody. Make it into a sheet and dry the sheet overnight at 37°C.

[0060] Step 2. Attach the sample pad and absorbent pad to the dried sheet to form a large card; the sample pad is made of glass fiber and contains hydrophilic surfactants, and the absorbent pad is made of filter paper.

[0061] Step 3. Cut the assembled card into test strips of the specified size, then assemble the test strips into the test plate's housing. The process is now complete. Figure 1 The test card shown is ready for use;

[0062] Step 4. Dilute the colloidal gold-antigen complex (hereinafter referred to as gold-labeled complex or gold label) to the required amount, then coat it into an incubation tube and dry or freeze-dry it.

[0063] Step 5. After drying or freeze-drying the incubation tubes in Step 4, heat-seal the openings with aluminum foil to complete the process. Figure 1 The incubation tubes shown are ready for use.

[0064] The test card in step 3 and the incubation tube in step 5 are used together to form the detection system described in this invention. It is then packaged in an aluminum foil bag.

[0065] II. Testing Steps

[0066] Detection Step 1. During detection, remove the test plate and incubation tube from the aluminum foil bag;

[0067] Detection step 2. Use one incubation tube for each sample. The bottom of the incubation tube has a dried gold label complex. If necessary, affix an appropriate label to each tube.

[0068] Detection step 3. (e.g.) Figure 5 As shown in Figure a, the seal 212 on the incubation tube 200 should be removed before adding the sample; as... Figure 5 As shown in Figure b, place the dropper vertically into the incubation tube and add 3 drops of the sample to be tested, 11; Figure 5 As shown in c and d, shake until the gold-labeled complex at the bottom is completely dissolved and mixed to form reconstituted solution 12; start the stopwatch and react for 5 minutes (if liquid adheres to the tube wall when adding buffer or sample, be sure to shake the reaction incubation tube to ensure the added solution reaches the bottom of the tube).

[0069] Detection step 4. Figure 5 As shown in e, place the new dropper vertically into the incubation tube and draw up the reconstituted product 12 from step 3;

[0070] Detection step 5. Figure 5 As shown in f, place the test plate on a flat and clean surface, and add 1 drop of buffer 304 and 2-3 drops of reconstitute 12 to the sample well of the test plate;

[0071] Step 6. Read the test results. If the test line shows color, there are no neutralizing antibodies in the sample. If the test line does not show color, there are neutralizing antibodies in the sample.

[0072] A drop of liquid dispensed from a dropper is approximately 20 to 30 microliters, for example, 25 microliters.

[0073] The order of adding the sample and buffer solution can be adjusted as needed. For example, in step 3, after adding the sample to the incubation tube with a dropper, two drops of buffer solution 304 are added to the incubation tube. Alternatively, step 3 could involve adding the sample to the incubation tube, shaking until the gold-labeled complex at the bottom is completely dissolved and mixed, reacting for a certain time, then aspirating the reconstituted solution from the incubation tube and adding it to the sample well of the test plate, while simultaneously adding buffer solution to the well. The reconstituted solution and buffer solution flow on the test strip, completing the detection. Another example: step 3 could involve adding the sample to the incubation tube, shaking until the gold-labeled complex at the bottom is completely dissolved and mixed, reacting for a certain time, then aspirating the reconstituted solution from the incubation tube and adding it to the sample well of the test plate. The reconstituted solution flow on the test strip, completing the detection.

[0074] Step 4 for the detection of the reconstituted material also employs other methods. For example, a dropper is attached to the opening of the incubation tube. After the gold-labeled complex at the bottom of the incubation tube is fully reconstituted, the bottom of the incubation tube is squeezed to mix the complex and the sample evenly. After the mixture reacts for 5 minutes, the incubation tube is squeezed, and the reconstituted material in the incubation tube is dropped into the test plate through the dropper.

[0075] In step 5 of the test, when adding the reconstituted solution to the sample well, air bubbles should be avoided as much as possible.

[0076] In step 6, which involves reading the test results, a preferred approach is to read the results after 15 minutes, and not after 30 minutes. The strength of the test line signal can be compared with the accompanying color chart to determine whether neutralizing antibodies are present in the sample.

[0077] Example 2 Sensitivity Test

[0078] Experimental group: This group uses the methods and equipment described in this invention to conduct testing. The invention... Figures 1 to 5 The detection system shown, as well as the preparation method and detection steps described in Example 1.

[0079] The control group: Testing was conducted using existing technology, employing a traditional test plate and testing procedure. The test strips in the traditional test plate were essentially the same as those in Example 1, except for the addition of a marking pad between the sample pad and the test pad. The marker from the incubation tube in the experimental group was transferred to this marking pad. The testing procedure involved directly adding the sample to be tested into the sample well of the test plate, allowing the sample to enter the sample pad of the test strip through the well.

[0080] The test plates and test strips used in both the experimental and control groups were from the same batch and from the same manufacturer. The reagents used in both groups were from the same batch and from the same manufacturer.

[0081] Samples: 24 positive samples were confirmed by ELISA to contain SARS-CoV-2 neutralizing antibodies, with sample numbers P-1 to P-24.

[0082] Table 1:

[0083] Sample number P-1 P-2 P-3 P-4 P-5 P-6 P-7 P-8 ELISA method inhibition rate 60％ 61％ 39％ 64％ 39％ 42％ 87％ 76％ Control group test results Negative Negative Negative Negative Negative Negative Positive Positive Experimental group test results Positive Positive Positive Positive Positive Positive Positive Positive Sample number P-9 P-10 P-11 P-12 P-13 P-14 P-15 P-16 ELISA method inhibition rate 36％ 96％ 91％ 55％ 53％ 41％ 89％ 90％ Control group test results Negative Positive Positive Negative Negative Negative Positive Positive Experimental group test results Positive Positive Positive Positive Positive Positive Positive Positive Sample number P-17 P-18 P-19 P-20 P-21 P-22 P-23 P-24 ELISA method inhibition rate 45％ 88％ 79％ 40％ 85％ 88％ 37％ 49％ Control group test results Negative Positive Positive Negative Positive Positive Negative Negative Experimental group test results Positive Positive Positive Positive Positive Positive Positive Positive

[0084] Note: An inhibition rate >30% is considered positive using the ELISA method.

[0085] The sensitivity test results are shown in Table 1. The test results demonstrate that the detection system and method described in this invention achieve higher sensitivity compared to existing technologies. The main difference in this invention is that the gold label, which is sprayed onto the marking pad in traditional test strips, is transferred to a separate incubation tube, allowing the gold label to incubate with the sample outside the test strip. This operation significantly improves the product's sensitivity.

[0086] Example 3

[0087] The detection system adopts Figure 1 , Figure 2 and Figure 3-2 The detection system shown, Figure 3-2 The test strip 120c includes an influenza A test line on its test pad, wherein the influenza A test line is coated with a qualified influenza A coated antibody 1.

[0088] In order to quickly screen out influenza A labeled antibody 2 that matches the qualified influenza A coated antibody 1, the influenza A labeled antibody 2 is labeled on qualified colloidal gold to form an antibody-gold label complex (i.e., the label). The complex solution is added to different incubation tubes according to different dilution concentrations. For example, if five concentrations of the same batch of raw materials need to be evaluated at one time, five incubation tubes are needed, and each incubation tube corresponds to one evaluation concentration.

[0089] For detailed operating procedures, please refer to... Figure 5-1 Before adding the sample, remove the seal 212 from the incubation tube 200, as shown below. Figure 5-1 As shown in Figure a; add buffer 304 dropwise to an incubation tube labeled with marker 303, as follows. Figure 5-1 As shown in b; add sample 11 containing influenza A virus to an incubation tube containing the labeled substance, as shown in Figure 1. Figure 5-1 As shown in c; the buffer, sample, and label are mixed to form reconstituted solution 12, as shown in the figure. Figure 5-1 As shown in d and e; after the required incubation time, use a clean pipette to remove the reconstituted product 12 from the incubation tube and add it to the sample well of the test plate 100, as shown. Figure 5-1 As shown in f and g. For example, to evaluate which of the five concentrations of the same batch of raw material is appropriate, the five concentrations of raw material can be placed in five different incubation tubes, according to... Figure 5-1 The procedure involves adding the reconstituted solution from five different incubation tubes to the sample wells of five test plates from the same batch. Then, based on the different color intensities of the detection lines on the test plates corresponding to the different incubation tubes, the appropriate evaluation concentration can be selected. The intensity of the detection line color can be obtained by comparing the detection line of the test plate with the color chart 500. Figure 12 As shown.

[0090] This operation allows for the simultaneous evaluation of multiple raw materials, different batches of the same raw material, and different concentrations of the same batch in a short period of time. It eliminates the need to spray the marker onto the binding pad and dry it before use, which greatly improves evaluation efficiency. On the other hand, since the large card used in the entire evaluation process does not require the application of a marking pad, assembly costs can be reduced, thereby reducing the overall evaluation cost.

Claims

1. A detection system comprising a test strip with lateral crossflow and an incubation tube, characterized in that, A marker that specifically binds to the analyte is placed in an incubation tube. A test strip is installed in a test plate, and the test plate has an incubation tube mounting base at the sample application well. The incubation tube is a tube with openings at both the top and bottom. The bottom of the incubation tube is mounted on the mounting base. During sample incubation in the incubation tube, or before the mixture of marker and sample is added to the sample pad through the sample application well of the test plate, the bottom opening of the incubation tube is not in communication with the liquid in the sample application well of the test plate. After incubation is complete and the incubated mixture is added to the sample pad, the bottom opening of the incubation tube becomes in communication with the liquid in the sample application well of the test plate. An O-ring is installed around the bottom opening of the incubation tube. After the incubation tube is mounted on the test plate, the bottom opening of the incubation tube is offset from the sample application well of the test plate, and the O-ring blocks the liquid flow between the outlet of the incubation tube and the sample application well. When the incubation tube is rotated, the bottom opening of the incubation tube aligns with the sample application well of the test plate, and the two become in communication, allowing the liquid in the incubation tube to flow into the test plate.

2. The detection system according to claim 1, characterized in that, The incubation tube and the test plate are two independent devices in the detection system.

3. The detection system according to claim 1, characterized in that, The test strip consists of, from upstream to downstream, a sample application pad, a detection pad, and an absorption pad, which are stacked on top of each other.

4. The detection system according to claim 3, characterized in that, The test strip also includes a flow guide pad, which is located between the sample application pad and the detection pad.

5. The detection system according to any one of claims 1 to 4, characterized in that, The test strip does not contain any markers that can specifically bind to the analyte.

6. A detection method for an analyte, characterized in that, The invention includes a detection system according to any one of claims 1 to 4; an incubation tube is installed on an incubation tube mounting base at the sample application port of a test plate, wherein the bottom opening of the incubation tube is not in communication with the liquid in the sample application port of the test plate; a sample is added to the incubation tube to form a mixture of sample and marker; and incubation is performed. Connect the bottom opening of the incubation tube to the liquid in the sample well of the test plate, and add the incubated mixture to the test strip; obtain the test result according to the signal indication of the test strip detection line.

7. The detection method according to claim 6, characterized in that, Add buffer solution to the incubation tube when adding the sample, or add buffer solution when adding the mixture to the test strip.

8. The detection method according to claim 6, characterized in that, The test strip does not contain any markers that can specifically bind to the analyte.

9. The detection method according to claim 6, characterized in that, The analyte is an antigen, antibody, or DNA / RNA aptamer.

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