Sample detection kit

Abstract

A sample detection kit, which comprises a detection test strip and sample treatment reagents, wherein the detection test strip comprises a sample pad, a hydrophobic pad, a labeling pad, and a detection pad which are connected in sequence; and the sample treatment reagents comprise two reagents: one of the sample treatment reagents is comprised in the sample pad, and the other sample treatment reagent is present in a reagent bottle. In the field of detection that requires sample pretreatment for sufficient release of an analyte in a sample, the kit simplifies operational steps, avoids erroneous results caused by improper manual operations, and exhibits a high detection sensitivity and a strong specificity.

Description

A sample detection kit Technical Field

[0001] This invention relates to the field of biological sample testing, and in particular to sample testing kits for which two sample processing solutions need to be temporarily mixed. Background Technology

[0002] Immunochromatographic assays, used to detect analytes in samples, have been widely applied since the 1880s. They are convenient, cost-effective, and readily available. This technique can be used to detect analytes in samples such as blood, saliva, urine, and lesion secretions to diagnose a patient's health. Based on the principle of specific binding of immune molecules, these molecules are solidified on a chromatographic support such as a PVC substrate or glass. Simultaneously, the immune molecules bind to groups such as latex, colloidal gold, fluorescent microspheres, and quantum dots, resulting in color development or excitation at specific wavelengths to display the detection results.

[0003] Immunochromatographic assays typically employ two methods: the double-antibody sandwich method and the competitive method, with the double-antibody sandwich method being the most widely used in antigen detection. For example, Chinese patent application CN202010893746.5 describes a test strip utilizing the double-antibody sandwich method for detecting Group A Streptococcus (Strep A). ​​A colored line appearing on the T-line in the test area indicates a positive result, meaning the analyte is present in the sample. Conversely, the absence of a colored line on the T-line indicates a negative result, meaning the analyte is not present in the sample.

[0004] However, existing test strips have some problems in practical applications for certain samples. For example, sample pretreatment is often required during testing to ensure the analyte is fully released. Therefore, sample pretreatment is an essential step for accurate results. In some tests, two sample processing solutions (solution A and solution B) need to be mixed in a specific ratio for pretreatment. However, pre-mixing these solutions makes them unsuitable for long-term storage, or the effectiveness of the pretreated sample decreases significantly after long-term storage. Furthermore, improper mixing ratios can lead to inaccurate results. Therefore, the two sample processing solutions are usually stored separately in two reagent bottles. However, storing the two solutions separately and packaging them with the test strip increases production steps; it also requires more space during packaging and storage, and is inconvenient for transportation. On the other hand, the process of mixing the two solutions before adding them to the sample and then applying them to the test strip is cumbersome and complex, increasing the risk of operational errors for ordinary operators. Summary of the Invention

[0005] To solve the above technical problems, the technical solution adopted by the present invention is as follows:

[0006] A test kit is provided, comprising a test strip and a sample processing solution. The test strip includes a sample pad, a label pad, and a test pad connected in sequence. The sample processing solution includes two reagents, with the sample pad containing one sample processing reagent and the other sample processing reagent stored in a reagent bottle.

[0007] When in use, it allows for sample pretreatment with only one sample processing reagent, eliminating the need for mixing two sample processing reagents in a specific ratio, thus simplifying the operation.

[0008] Furthermore, the test strip also includes a hydrophobic pad, which is connected between the sample pad and the marker pad.

[0009] There is a hydrophobic pad between the sample pad and the label pad, which slows down the speed at which the sample passes through the hydrophobic pad. This allows the sample to be fully processed on the sample pad, thereby fully releasing the analytes in the sample and increasing the concentration of the analytes, thus maintaining the detection sensitivity of the test strip.

[0010] Furthermore, the hydrophobic pad is made of a hydrophobic polyester film.

[0011] Furthermore, the hydrophobic pad is made of polyester film and contains EPS, sucrose, and starch.

[0012] Furthermore, the hydrophobic pad is made of polyester film and is treated with a hydrophobic treatment solution; the hydrophobic pad treatment solution contains EPS, sucrose and starch.

[0013] Furthermore, the hydrophobic treatment solution contains 5g / L EPS, 25g / L sucrose, 10g / L starch, and has a pH value of 8.0.

[0014] Furthermore, the sample detection kit is used for the detection of Group A Streptococcus and Group B Streptococcus.

[0015] Furthermore, the sample processing reagent on the sample pad contains sodium nitrite, and the sample processing reagent in the reagent bottle contains glacial acetic acid.

[0016] Furthermore, the sample treatment reagent on the sample pad is a 4M aqueous solution of sodium nitrite, and the sample treatment reagent in the reagent bottle is a 2M aqueous solution of glacial acetic acid.

[0017] Furthermore, the sample testing kit is used for the detection of Chlamydia trachomatis.

[0018] Furthermore, the sample treatment reagent on the sample pad contains sodium hydroxide, and the sample treatment reagent in the reagent bottle contains tartaric acid.

[0019] Furthermore, the sample treatment reagent on the sample pad is a 0.5M aqueous solution of sodium hydroxide, and the sample treatment reagent in the reagent bottle is a 0.5M aqueous solution of tartaric acid.

[0020] Furthermore, the test strip also includes an absorbent pad and a bottom PVC base plate. The sample pad, and / or hydrophobic pad, marking pad, test pad, and absorbent pad are all connected to the PVC base plate.

[0021] The advantages of this invention compared to the prior art are:

[0022] First, in the test strip preparation stage, this invention treats the sample pad with the active ingredient of one of the two sample processing reagents. Therefore, when using the test strip, the sample is pretreated directly with one sample processing reagent, eliminating the need for mixing the two sample processing reagents in a specific ratio, thus simplifying the operation.

[0023] Secondly, the sample pad treatment involves adding one of the active ingredients of the two sample treatment reagents, eliminating the need to mix the two reagents in a specific ratio. This avoids inaccurate test results due to incorrect mixing ratios of the two reagents.

[0024] Third, the sample pad treatment involves adding one of the two sample treatment reagents' active ingredients, eliminating the need for one sample treatment reagent filling step, reducing production processes, improving production efficiency, and lowering production costs.

[0025] Fourth, by treating the active ingredient of one of the two sample processing reagents in the sample pad, one reagent bottle for storing the processing reagent is eliminated, which saves costs and further saves packaging space for the test kit, reducing storage and transportation costs.

[0026] Fifth, the present invention adds a hydrophobic pad between the sample pad and the label pad, which slows down the flow rate of the sample when it passes through the hydrophobic pad. This allows the sample to be fully processed on the sample pad before flowing into the label pad, so that the analytes in the sample are fully released. This reduces the sample processing reagent while maintaining the detection sensitivity of the test strip. Attached Figure Description

[0027] Figure 1 is a schematic diagram of the operation steps of existing conventional Group A Streptococcus (Strep A) or Group B Streptococcus (Strep B) detection kits.

[0028] Figure 2 is a schematic diagram of the structure of existing conventional test strips.

[0029] Figure 3 is a schematic diagram of the structure of the test strip of the present invention.

[0030] Figure 4 is a schematic diagram of a test plate equipped with the test strip of the present invention.

[0031] Figure 5 is a schematic diagram of the operation steps of the detection kit of the present invention. Detailed Implementation

[0032] Embodiments of the present invention are described in detail below, wherein the same or similar reference numerals denote the same or similar elements or elements with similar functions throughout. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention and should not be construed as limiting the present invention.

[0033] The numbering of steps mentioned in the various embodiments is merely for descriptive convenience and does not imply a sequential relationship. Different steps in various specific embodiments can be combined in different orders to achieve the inventive objective of this invention.

[0034] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0035] As shown in Figure 1, a standard test kit includes two sample processing reagents, reagent A and reagent B, stored in a reagent bottle. The operating steps of a standard test kit are shown in steps a to f of Figure 1, including: During testing, reagents A and B are mixed in a specific ratio. A swab is inserted into the mixture to pre-treat the sample, ensuring the analyte is fully released. The solution treated with the mixed reagents is then dropped onto a sample pad. The sample pad collects the treated sample solution and filters out impurities that may affect the test results. Specific molecules labeled with chromogenic groups on the label pad bind to the analyte, forming a chromogenic group-specific molecule-analyte complex. The test pad is typically a nitrocellulose membrane immobilizing the specific molecules, which captures the chromogenic group-specific molecule-analyte complex, resulting in a test line indicating the result on the test pad.

[0036] As shown in Figure 2, a conventional test strip consists of a sample pad 1, a marking pad 2, a detection pad 4, an absorbent pad 5, and a bottom PVC base plate 6. The sample pad 1, marking pad 2, detection pad 4, and absorbent pad 5 are typically assembled on the PVC base plate 6. Generally, during test strip preparation, the detection pad 4 is placed first on the bottom PVC base plate. One downstream end of the detection pad 4 is connected to the absorbent pad 5, and the upstream end is connected to the marking pad 2. The upstream end of the marking pad 2 is connected to the sample pad 1. In specific designs, the length of the PVC base plate is typically between 4cm and 12cm, and the width is between 2mm and 10mm. In one specific embodiment, the length is 6cm and the width is 3.5mm.

[0037] In practice, two sample processing reagents, one (e.g., reagent A) and the other (e.g., reagent B), are mixed in a specific ratio to pretreat the sample. The pretreated sample solution is then dropped onto the sample pad. The sample solution is absorbed into the sample pad and flows laterally along the test strip. The analyte in the sample solution passes through the label pad and reacts with specific molecules in the label pad to form a complex. This complex continues to flow along the test strip to the detection line 41 (T line) and control line 42 (C line) on the test pad, where it reacts to produce colored lines indicating the test result. Excess sample solution continues to flow along the test strip downstream of the test pad and is absorbed by the absorbent pad, thus completing the test.

[0038] In practice, conventional test kits require mixing two sample processing solutions, reagent A and reagent B, which is cumbersome. Furthermore, the samples need to be processed within a short time (e.g., 1 hour) after mixing; otherwise, the effectiveness of the mixed sample processing solution will decrease, affecting the sensitivity of the test strip. Incorrect mixing ratios can also easily lead to erroneous test results.

[0039] Analysis revealed that the substances in the two sample processing reagents, A and B, cannot be stored together for long periods and must be mixed in precise proportions; adding too much or too little reagent A or B will affect the detection results. Using reagent A or reagent B alone does not effectively release the analyte, leading to low sensitivity. For example, in Example 1, the active ingredient in reagent A is sodium nitrite, and the active ingredient in reagent B is acetic acid. When mixed, reagents A and B react to produce nitrite and sodium acetate. Nitrite can effectively degrade the cell walls of Group A Streptococcus, allowing the analyte to be fully released. However, nitrite is chemically unstable in solution and gradually decomposes, significantly reducing the effectiveness of nitrite solutions in treating samples after long-term storage. In Example 2, the principle of sample processing with reagents A and B is similar to Example 1, except that they degrade the cell walls of Group B Streptococcus, resulting in the full release of the analyte. In Example 3, the active ingredient of reagent A is sodium hydroxide, and the active ingredient of reagent B is tartaric acid. The sample is first treated in an acidic environment, and then sodium hydroxide is added to treat the sample to ensure that the analyte is fully released.

[0040] To solve this problem, it is necessary to ensure that the active ingredients in reagents A and B react to fully release the analyte from the sample. Simultaneously, a simple operating procedure is required to facilitate the correct reaction ratio between the active ingredients in reagents A and B. Therefore, an improved test strip of the present invention includes a sample pad containing one sample processing reagent and another sample processing reagent stored in a reagent bottle. For example, the sample pad contains reagent A, and the reagent bottle contains reagent B. In use, simply add the other sample processing reagent, such as reagent B, from the reagent bottle, after mixing it with the sample, to the sample pad. This allows the sample processing reagent on the sample pad, such as reagent A, to mix with reagent B and simultaneously with the sample. This achieves both timely mixing of reagents A and B and the sample, and also ensures mixing in a fixed ratio.

[0041] As shown in Figure 3, the improved test strip 10 comprises a sample pad 1, a hydrophobic pad 3, a marking pad 2, a detection pad 4, an absorbent pad 5, and a bottom PVC base plate 6. Based on a conventional test strip, the reagent bottle containing reagent A from the sample processing reagents is omitted, while the sample pad 1 contains a certain amount of the effective component of reagent A. A hydrophobic pad 3 is added between the sample pad 1 and the marking pad 2. The operation steps of the test kit described in this invention are shown in steps a to d in Figure 5, including: pretreating the sample with reagent B, then dropping the treated sample solution onto sample pad 1. The sample solution is adsorbed into the sample pad. The hydrophobic pad 3 at the downstream end of the sample pad needs a certain amount of time to be wetted. Before being wetted, the sample solution is blocked by the hydrophobic pad 3, causing the sample solution to accumulate on sample pad 1. The sample pad contains a certain amount of the active ingredient of reagent A, which reacts fully with the sample solution, allowing the analyte in the sample to be fully released. Then, the hydrophobic pad begins to be wetted. The fully released analyte passes through the hydrophobic pad to the label pad, reacts with the specific molecules in the label pad 2 to form a complex, and continues to flow along the test strip to the detection line 41 (T line) and control line 42 (C line) on the test pad to react, thereby displaying the test result by color lines. Excess sample solution continues to flow along the test strip to the downstream end of the test pad and is absorbed by the absorbent pad, finally completing the test.

[0042] As shown in Figure 4, the test plate 20 includes an upper cover 20, a lower plate 21, and a test strip 10 installed between the upper cover and the lower plate. Depending on the needs, either the test strip or the test plate can be used for sample testing. The test kit includes test strips and sample processing reagents; alternatively, the test kit may include a test plate and sample processing reagents.

[0043] Example 1: Comparison of conventional test strips and improved test strips that incorporate reagent A into the sample pad and add a hydrophobic pad for detecting group A streptococci.

[0044] Material:

[0045] Sampling swabs;

[0046] The standard test strips are products of Hangzhou Lin'an Aikon Biotechnology Co., Ltd.

[0047] The improved test strip refers to the test strip of the present invention in which reagent component A is treated in the sample pad and a hydrophobic pad is added between the sample pad and the label pad.

[0048] The control group test strips refer to test strips in which reagent A is treated in the sample pad.

[0049] The sample was an inactivated strain of Group A Streptococcus.

[0050] step:

[0051] 1. Prepare inactivated Group A Streptococcus strains. Use PBS to prepare 2*10... 8 Group A Streptococcus inactivated strains were serially diluted to 2 x 10⁻⁶ CFU / ml. 7 CFU / ml, 2*10 6 CFU / ml, 2*10 5 CFU / ml, 2*10 4 CFU / m.

[0052] 2. Positive sample preparation. Place 20 μL of the prepared sterilized bacterial strain into the head of the sampling swab to prepare gradient positive swabs with different concentration gradients. Positive swab 1 (containing 20 μL of 2*10...) 7 CFU / ml strain), 2 positive swabs (containing 20ul 2*10 6 CFU / ml strain), 3 positive swabs (containing 20ul 2*10 5 CFU / ml strain), 4 positive swabs (containing 20ul 2*10 4 (CFU / ml strain).

[0053] 3. Negative sample preparation. Pharyngeal secretions were collected from different individuals using sampling swabs.

[0054] 4. Sample preprocessing.

[0055] For standard test strips: First, mix 8 drops (about 200ul) each of reagent A and reagent B. Then, put the prepared positive or negative sample into the mixture of reagent A and reagent B, rotate and stir 15 times, let stand for 1 minute, and then discard the swab.

[0056] Improved test strip: Place the prepared positive or negative sample into reagent B, rotate and stir 15 times, let stand for 1 minute, and then discard the swab.

[0057] 5. Sample addition and testing. Add 4 drops (approximately 100 μL) of the pretreated sample to the sample pad. Compare the color intensity on the test pad with the colorimetric chart on the test strip to obtain the numerical value.

[0058] 6. Standard test strips are prepared according to the following method:

[0059] 6.1 Preparation of sample processing reagents, namely reagent A and reagent B. Reagent A is a 4M aqueous solution of sodium nitrite, and reagent B is a 2M aqueous solution of glacial acetic acid.

[0060] 6.2 Preparation of the test pad. The test pad uses a nitrocellulose membrane as a carrier, with two lines on the NC membrane: the test line (T line) and the control line (C line). The test line (T line) is immobilized with anti-Group A Streptococcus antibody-1, and the control line (C line) is immobilized with goat anti-rabbit IgG polyclonal antibody. The T line and C line are immobilized on the NC membrane using a membrane application machine. The concentration of anti-Group A Streptococcus antibody-1 in the T line solution is 0.5 mg / ml, and the dilution buffer is borate buffer with a pH of 8.0. The concentration of goat anti-rabbit IgG polyclonal antibody in the C line solution is 2.0 mg / ml, and the dilution buffer is borate buffer with a pH of 8.0. After processing by the membrane application machine, the test pad is dried in a 37°C oven.

[0061] 6.3 Preparation of Labeled Pads. The labeled pads used glass fiber as a carrier. The labeling solution was applied to the glass fiber using a spray gun. The labeling solution was obtained by covalently coupling anti-Group A Streptococcus antibody-2 with latex microspheres. The dilution buffer was 0.8% BSA, pH 8.0. After spray gun processing, the labeled pads were dried in a 37°C oven.

[0062] 6.4 Sample Pad Preparation. Glass fiber was used as the carrier for the sample pads. The sample pad treatment solution was sprayed onto the glass fiber. The treatment solution consisted of: 20 g / L borate, 5 g / L calcium dodecylbenzene xanthate, 10 g / L PVP, and 10 g / L BSA, with a pH of 8.0. The treated sample pads were then dried in a 37°C oven.

[0063] 6.5 Assembly of the test strip. Assemble the prepared components in the order shown in Figure 2 for a standard test strip. First, assemble the detection pad, with one end of the absorbent pad covering the downstream end of the detection pad, the downstream end of the labeling pad covering the upstream end of the detection pad, and the downstream end of the sample pad covering the upstream end of the labeling pad.

[0064] 7. The improved test strip, which incorporates reagent A into the sample pad and adds a hydrophobic pad (i.e., the improved test strip), is prepared according to the following method:

[0065] 7.1 Preparation of sample processing reagent, i.e., reagent B: Reagent B is the same as reagent B in regular test strips.

[0066] 7.2 The preparation methods for the test pad and the label pad are the same as those for conventional test strips.

[0067] 7.3 Sample pad preparation: Compared with conventional sample pads, only 4M sodium nitrite (Reagent A) was added to the sample pad treatment solution, while the other components and treatment methods remained unchanged.

[0068] 7.4 Preparation of hydrophobic pad: The hydrophobic pad uses a polyester film as a carrier. The hydrophobic pad treatment solution is sprayed onto the polyester film. The hydrophobic pad treatment solution is: EPS 5g / L, sucrose 25g / L, starch 10g / L, pH 8.0.

[0069] 7.5 Test strip assembly. Assemble the processed components in the order shown in Figure 3 of the improved test strip. First, assemble the test pad. One end of the absorbent pad covers the downstream end of the test pad, the downstream end of the marker pad covers the upstream end of the test pad, the downstream end of the hydrophobic pad covers the upstream end of the marker pad, and the downstream end of the sample pad covers the upstream end of the hydrophobic pad.

[0070] Interpretation of test results:

[0071] Colorimetric chart: The colorimetric chart has a color rendering range of 0-10, i.e., L0-L10. The color intensity increases progressively from 0 to 10. When the color intensity of the test line (T line) is between L3 (inclusive) and L10, the test result is positive. When the color intensity of the test line (T line) is between L0 and L3 (exclusive), the test result is negative.

[0072] 8. The control group test strips were prepared according to the following method:

[0073] The preparation method of the control group test strip is basically the same as that of "7. Improved test strip with reagent A component treated in the sample pad and an additional hydrophobic pad (i.e., improved test strip) prepared according to the following method", except that no hydrophobic pad is placed between the sample pad and the label pad.

[0074] Table 1: Detection Results of Group A Streptococcus Positive Samples

[0075] Table 2: Negative Sample Test Results

[0076] Analysis of test results:

[0077] Table 1 shows that mixing reagents A and B 4 hours in advance resulted in a 10-fold decrease in sensitivity. Test strips with reagent A pre-treated sample pads exhibited sensitivity between those using conventional test strips with reagents A and B mixed within 1 hour and those using conventional test strips with reagents A and B mixed 4 hours in advance. The improved test strips, with reagent A pre-treated sample pads and an added hydrophobic layer, showed the same sensitivity performance as conventional test strips with reagents A and B mixed within 1 hour, without any negative impact on sensitivity.

[0078] The results in Table 2 show that the improved test strip has the same specificity as the conventional test strip, and its specificity performance was not negatively affected.

[0079] Without compromising sensitivity and specificity, the improved test kit eliminates the need for a reagent bottle for storing reagent A and the step of mixing reagent A and reagent B, simplifying operation, reducing production steps, increasing production efficiency, and lowering production costs.

[0080] Example 2: Comparison of conventional test strips and improved test strips with reagent A component treated in the sample pad and an additional hydrophobic pad for detecting group B streptococci.

[0081] 1. The operation steps in this embodiment are the same as those in Example 1, except that the strain used for preparing the positive sample is different, and the labeling solution and the antibody for the T-line detection line are different.

[0082] 2. Prepare inactivated Group B Streptococcus strains. Use PBS to prepare 3.8*10 8 Group A Streptococcus inactivated strains were serially diluted to 3.8 x 10⁻⁶ CFU / ml. 7 CFU / ml, 3.8*10 6 CFU / ml, 3.8*10 5 CFU / ml, 3.8*10 4 CFU / ml.

[0083] 3. Positive Sample Preparation. Place 20 μL of the prepared sterilized bacterial strain into the head of the sampling swab to prepare gradient positive swabs with different concentration gradients. Positive swab 1 (containing 20 μL of 3.8*10...) 7 CFU / ml strain), 2 positive swabs (containing 20ul 3.8*10 6 CFU / ml strain), 3 positive swabs (containing 20ul 3.8*10 5 CFU / ml strain), 4 positive swabs (containing 20ul 3.8*10 4 (CFU / ml strain).

[0084] 4. The detection line T is immobilized with anti-Group B Streptococcus antibody-1. The labeling solution can be obtained by covalently coupling anti-Group B Streptococcus antibody-2 with latex microspheres. The remaining procedures are the same as in Example 1.

[0085] Table 3: Detection Results of Group B Streptococcus Positive Samples

[0086] Table 4: Negative Sample Test Results

[0087] Analysis of test results:

[0088] Table 3 shows the same results: mixing reagents A and B 4 hours in advance resulted in a 10-fold decrease in sensitivity. The test strip with reagent A pre-treated sample pad exhibited sensitivity between that of a conventional test strip used within 1 hour of mixing reagents A and B, and a conventional test strip with reagents A and B mixed 4 hours in advance. The improved test strip, with reagent A pre-treated sample pad and an added hydrophobic layer, showed slightly higher sensitivity than the conventional test strip (used within 1 hour of mixing reagents A and B), without negatively impacting sensitivity performance.

[0089] Table 4 shows that the improved test strip has the same specificity as the conventional test strip, and its specificity performance was not negatively affected.

[0090] Without compromising sensitivity and specificity, the improved test kit eliminates the need for a reagent bottle for storing reagent A and the step of mixing reagent A and reagent B, simplifying operation, reducing production steps, increasing production efficiency, and lowering production costs.

[0091] Example 3: Comparison of conventional test strips and improved test strips with reagent A component treated in the sample pad and an additional hydrophobic pad for detecting Chlamydia trachomatis.

[0092] Except for the following steps, which are different from those in Example 1, all other operation steps in this embodiment are the same as those in Example 1.

[0093] 1. Standard test strips

[0094] 1.1 The sample processing solutions are reagent A and reagent B. Reagent A is a 0.5M aqueous solution of sodium hydroxide, and reagent B is a 0.5M aqueous solution of tartaric acid.

[0095] 1.2 Sample pretreatment. The sample was placed in 4 drops of reagent A and stirred 15 times, then left to stand for 1 minute. Then 4 drops of reagent B were added and stirred 15 times, left to stand for 1 minute, and then the swab was discarded.

[0096] 1.3 Prepare the Chlamydia trachomatis inactivation quality control sample. Use PBS to prepare 4.5*10 7Copies / ml of Chlamydia trachomatis inactivated quality control sample were serially diluted to 4.5 x 10^6 copies. 6 copies / ml, 4.5*10 5 copies / ml, 4.5*10 4 copies / ml.

[0097] 1.4 Preparation of positive samples. Place 20 μL of the prepared inactivated quality control sample into the head of the sampling swab to prepare gradient positive swabs with different concentration gradients. Positive swab 1 (containing 20 μL of 4.5*10...) 6 CFU / ml strain), 2 positive swabs (containing 20ul 4.5*10 5 CFU / ml strain), 3 positive swabs (containing 20ul 4.5*10 4 (CFU / ml strain).

[0098] 1.5 Negative Sample Preparation. Using sampling swabs, cervical swabs were collected from women and urethral swabs from men from different individuals.

[0099] 1.6 The T-line of the detection line is immobilized with anti-Chlamydia trachomatis lipopolysaccharide monoclonal antibody-1. The labeling solution can be obtained by covalently coupling anti-Chlamydia trachomatis lipopolysaccharide monoclonal antibody-2 with latex microspheres. The remaining procedures are the same as in Example 1.

[0100] 2. Improved test strip

[0101] 2.1 Sample preprocessing. Same as in Example 1.

[0102] 2.2 Preparation of sample processing solution, i.e., reagent B: Reagent B is the same as reagent B in conventional test strips.

[0103] 2.3 The preparation methods for the test pad and the label pad are the same as those for conventional test strips.

[0104] 2.4 Sample pad preparation: Compared with conventional sample pads, only 0.5M sodium hydroxide (reagent A) was added to the sample pad treatment solution, pH 12.0, while the other components and treatment methods remained unchanged.

[0105] Table 5: Detection Results of Positive Samples for Chlamydia trachomatis

[0106] Table 6: Negative Sample Test Results

[0107] Analysis of test results:

[0108] Table 5 shows that the improved test strip, which incorporates reagent A into the sample pad and adds a hydrophobic layer, performs the same as the conventional test strip (used within 1 hour of mixing reagent A and reagent B), with no negative impact on sensitivity. The conventional test strip, which pre-mixes reagent A and reagent B, and the test strip with reagent A incorporated into the sample pad, exhibit significantly reduced sensitivity.

[0109] Table 6 shows that the improved test strip has the same specificity as the conventional test strip, and its specificity performance was not negatively affected.

[0110] In this way, without reducing sensitivity and specificity, the improved test kit eliminates the need for a reagent bottle to store reagent A and the step of mixing reagent A and reagent B, simplifying operation, reducing production steps, increasing production efficiency, and reducing production costs.

[0111] The above description is only a preferred embodiment of the present invention. It should be noted that those skilled in the art can make several improvements and modifications without departing from the concept of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A sample detection kit, comprising test strips and sample processing reagents, characterized in that, The test strip includes a sample pad, a label pad, and a test pad connected in sequence; the sample processing reagent includes two reagents, one of which is included in the sample pad, and the other is contained in a reagent bottle; the test strip also includes a hydrophobic pad, which is connected between the sample pad and the label pad.

2. The sample detection kit according to claim 1, characterized in that, The hydrophobic pad is made of hydrophobic polyester film.

3. The sample detection kit according to claim 1, characterized in that, The hydrophobic pad is made of polyester film and is treated with a hydrophobic treatment solution containing EPS, sucrose, and starch.

4. The sample detection kit according to claim 3, characterized in that, The hydrophobic pad treatment solution contains 5g / L EPS, 25g / L sucrose, and 10g / L starch, with a pH of 8.

0.

5. The sample detection kit according to claim 1 or 3, characterized in that, The sample detection kit is used for the detection of Group A Streptococcus and Group B Streptococcus.

6. The sample detection kit according to claim 5, characterized in that, The sample treatment reagent on the sample pad contains sodium nitrite, and the sample treatment reagent in the reagent bottle contains glacial acetic acid.

7. The sample detection kit according to claim 6, characterized in that, The sample treatment reagent on the sample pad is a 4M aqueous solution of sodium nitrite, and the sample treatment reagent in the reagent bottle is a 2M aqueous solution of glacial acetic acid.

8. The sample detection kit according to claim 1 or 3, characterized in that, The sample detection kit is used for the detection of Chlamydia trachomatis.

9. The sample detection kit according to claim 8, characterized in that, The sample treatment reagent on the sample pad contains sodium hydroxide, and the sample treatment reagent in the reagent bottle contains tartaric acid.

10. The sample detection kit according to claim 9, characterized in that, The sample treatment reagent on the sample pad is a 0.5M aqueous solution of sodium hydroxide, and the sample treatment reagent in the reagent bottle is a 0.5M aqueous solution of tartaric acid.

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