Colloidal gold detection test strip for mycoplasma pneumoniae as well as preparation method and application of colloidal gold detection test strip

Abstract

The invention relates to a colloidal gold detection test strip for mycoplasma pneumoniae as well as a preparation method and application of the colloidal gold detection test strip, and belongs to the technical field of immunodetection. A colloidal gold labeled mycoplasma pneumoniae detection antibody reconstitution fluid and a quality control detection antibody reconstitution fluid are prepared, mixed and sprayed on the combination pad, solutions containing capture antibodies with specific concentrations are sprayed on the detection line and the quality control line arranged on the coating film respectively, and finally all the components are assembled. The test strip has the advantages of high sensitivity and specificity, low detection limit, simplicity, convenience and rapidness in operation, capability of obtaining a result in a short time, and suitability for on-site instant detection, provides a reliable means for diagnosis of mycoplasma pneumoniae infection, and has a wide application prospect in the field of auxiliary diagnosis of pneumonia.

Description

Technical Field

[0001] The invention relates to the technical field of immunoassay detection, and in particular to a colloidal gold test strip for detecting mycoplasma pneumoniae, a preparation method and application thereof. Background Art

[0002] Mycoplasma pneumoniae, as a pathogen that frequently appears in the field of respiratory diseases, plays a major role in causing community-acquired pneumonia. Especially in children and adolescents, the infection rate is relatively high, and it is one of the important pathogenic factors of respiratory infectious diseases in this age group. This microorganism has a unique cell structure, and its most notable feature is the lack of a cell wall. The lack of a cell wall makes Mycoplasma pneumoniae show natural resistance to many conventional antibiotics. This characteristic poses a huge challenge to clinical treatment, because traditional antibiotics that target cell wall synthesis or act on cell wall-related targets are not effective against it. Therefore, in the response to Mycoplasma pneumoniae infection, rapid and accurate diagnosis has become a crucial link. Only by timely and accurate determination of the infection can a solid basis be provided for subsequent effective treatment and epidemic prevention and control, and to avoid aggravation of the disease or expansion of the scope of spread due to delayed diagnosis.

[0003] Traditional methods for detecting Mycoplasma pneumoniae include serological tests, PCR technology, and culture methods. Serological tests are mainly based on detecting specific antibodies produced against Mycoplasma pneumoniae in patients, and the infection status is determined by detecting the presence and titer changes of antibodies in blood samples. However, this method is extremely complicated to operate, requiring the collection of patient blood samples and a series of complex laboratory processing steps, including serum separation, preparation and incubation of antibody detection reagents, multiple washing operations, etc. The entire process is time-consuming and usually takes several hours or even days to obtain preliminary results. Moreover, the accuracy of serological tests is easily affected by various factors, such as the patient's infection time, individual immune response differences, etc., which may lead to false positive or false negative results.

[0004] PCR, or polymerase chain reaction technology, is highly sensitive and specific and can detect very small amounts of pathogen nucleic acid. However, PCR technology has extremely high professional requirements for laboratory equipment and technicians. Laboratories need to be equipped with expensive instruments such as high-precision thermal cyclers, nucleic acid extraction equipment, and professional fluorescent quantitative PCR instruments. At the same time, operating technicians must undergo rigorous professional training and be proficient in complex technical links such as nucleic acid extraction, primer design, PCR reaction system preparation, amplification program settings, and result analysis. In addition, the PCR detection process involves multiple steps, including sample collection, transportation, storage, nucleic acid extraction, amplification reaction, and result interpretation. Each link requires strict quality control, and the slightest carelessness may lead to deviations in the results. Moreover, the high cost of PCR testing, reagent costs, equipment maintenance costs, and labor costs make it difficult for this detection method to be widely carried out in primary medical institutions, limiting its popularization and application in clinical diagnosis.

[0005] The culture method is a traditional classic method for detecting Mycoplasma pneumoniae. Patient samples are inoculated into a specific culture medium, and after culturing for a period of time under appropriate culture conditions, the growth of Mycoplasma pneumoniae is observed. However, the disadvantages of this method are very obvious. First, the culture method is extremely time-consuming, and it generally takes several days to weeks to obtain clear culture results, which is unacceptable for patients who are in urgent need of timely diagnosis and treatment. Secondly, the culture method has strict requirements on the culture environment and conditions. It is necessary to strictly control multiple factors such as the composition, pH value, temperature, and humidity of the culture medium. At the same time, it is also necessary to prevent contamination by other bacteria, otherwise it is easy to cause culture failure. Furthermore, the culture method is difficult to operate, and requires professional microbiology technicians to operate, and requires rich experience and skills, which is often difficult to meet in primary medical institutions. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides a colloidal gold test strip for detecting Mycoplasma pneumoniae, wherein a sample pad, a binding pad, a coating film and a water absorbent pad are sequentially formed on a bottom plate, a Mycoplasma pneumoniae detection line and a quality control line are arranged on the coating film, and a colloidal gold test strip for detecting Mycoplasma pneumoniae with high sensitivity, high specificity and low detection limit is prepared.

[0007] The first object of the present invention is to provide a method for preparing a colloidal gold test strip for Mycoplasma pneumoniae, the preparation method specifically comprising the following steps:

[0008] (1) Soaking the sample pad in a sample pad treatment solution, wherein the sample pad treatment solution comprises a buffer, polyvinyl pyrrolidone, a surfactant, a preservative and a blocking agent, the pH value of the sample pad treatment solution is 7-8, and the amount of the sample pad treatment solution added is 0.05-0.1 mL / cm 2 ;

[0009] (2) taking a colloidal gold solution, adding a Mycoplasma pneumoniae detection antibody and bovine serum albumin to make the final concentration of the Mycoplasma pneumoniae detection antibody be 13-18 μg / mL, centrifuging to obtain a precipitate, adding a colloidal gold marker reconstitution solution to the precipitate to obtain a colloidal gold-labeled Mycoplasma pneumoniae detection antibody reconstitution solution, wherein the concentration of the colloidal gold solution is OD8-10;

[0010] Take the colloidal gold solution, add the quality control detection antibody and bovine serum albumin to make the final concentration of the quality control detection antibody be 8-12 μg / mL, centrifuge to obtain the precipitate, add the colloidal gold marker re-solution to the precipitate, and obtain the colloidal gold-labeled quality control detection antibody re-solution;

[0011] The colloidal gold marker complex solution includes Tris-HCL, casein or bovine serum albumin, sucrose, trehalose, sodium azide or ProClin300;

[0012] The colloidal gold-labeled Mycoplasma pneumoniae detection antibody reconstitution solution and the colloidal gold-labeled quality control detection antibody reconstitution solution are evenly mixed to prepare a mixed reconstitution solution, and the mixed reconstitution solution is sprayed on the conjugate pad;

[0013] (3) Setting a test line and a quality control line on the coating membrane, and spraying a solution containing Mycoplasma pneumoniae capture antibody and a solution containing quality control capture antibody respectively;

[0014] The final concentration of the Mycoplasma pneumoniae capture antibody is 0.6-1 mg / mL, and the final concentration of the quality control capture antibody is 0.8-1.2 mg / mL;

[0015] (4) Assembling the sample pad, the binding pad, the coating membrane and the water-absorbing pad to obtain the Mycoplasma pneumoniae colloidal gold test strip.

[0016] Furthermore, the method further includes a step of pretreating the conjugate pad, wherein the pretreatment is to immerse the conjugate pad in a conjugate pad treatment solution, wherein the conjugate pad treatment solution is composed of 40-80 mM Tris-HCl, 0.4-0.8 wt % casein or bovine serum albumin, 0.8-1.5 wt % polyvinyl pyrrolidone K30, 0.04-0.08 wt % Tween 20, and 0.02-0.06 wt % sodium azide or ProClin300.

[0017] Furthermore, the addition amount of the conjugate pad treatment solution is 0.04-0.08 mL / cm 2 .

[0018] Furthermore, in step (1), the sample pad treatment solution consists of 40-80 mM Tris-HCl, 0.4-0.8 wt % casein or BSA, 0.8-1.5 wt % polyvinylpyrrolidone K30, 0.8-1.5 wt % surfactant S9, 0.04-0.1 wt % Tween 20, 0.02-0.05 wt % sodium azide or ProClin300 and 0.002-0.006 wt % heterophilic antibody blocker HBR.

[0019] Furthermore, in step (2), the concentration of the colloidal gold-labeled Mycoplasma pneumoniae detection antibody in the mixed reconstitution solution is OD45-70, and the concentration of the colloidal gold-labeled quality control detection antibody is OD20-30.

[0020] Furthermore, in step (2), the spraying amount of the mixed solution is 2.5-4 μL / cm.

[0021] Furthermore, in step (2), the colloidal gold marker complex solution includes 50-100 mM Tris-HCl, 0.4-0.8 wt % casein or BSA, 15-25 wt % sucrose, 4-10 wt % trehalose, and 0.02-0.06 wt % sodium azide or ProClin300.

[0022] Furthermore, the pH value of the colloidal gold marker complex solution is 8-8.5.

[0023] Furthermore, in step (3), the spraying amount of the solution containing the Mycoplasma pneumoniae capture antibody on the detection line is 1-2 μL / cm.

[0024] The second object of the present invention is to provide a colloidal gold test strip for Mycoplasma pneumoniae, which is prepared using any of the above methods.

[0025] Furthermore, the Mycoplasma pneumoniae colloidal gold test strip also includes a sample diluent.

[0026] Furthermore, the sample diluent consists of 1-1.5wt% Tris, 0.5-1wt% BSA or casein, 0.5-1wt% surfactant S22, and 0.03-0.06wt% sodium azide or ProClin300.

[0027] Furthermore, the pH value of the sample diluent is 7.0-7.5.

[0028] The third object of the present invention is to provide the use of the colloidal gold test strip in the preparation of pneumonia auxiliary diagnosis products.

[0029] Beneficial effects of the present invention:

[0030] The colloidal gold test strip for Mycoplasma pneumoniae provided by the present invention has many significant technical effects. It has excellent performance in terms of sensitivity and specificity. By optimizing the antibody labeling and coating process, it can accurately identify Mycoplasma pneumoniae, greatly reducing the probability of false positive and false negative results, thereby providing a highly reliable basis for clinical diagnosis. Its detection limit is as low as 10 3 CFU / mL can effectively detect extremely low concentrations of Mycoplasma pneumoniae antigens, which is of vital importance for the early diagnosis of Mycoplasma pneumoniae infection, helping to detect the disease in time at the early stage of infection and to provide patients with the best treatment opportunity. In terms of operation, the test strip is simple and fast, does not rely on complex instruments and equipment, and only requires a simple operation process to complete the test in a short time. It is very suitable for on-site instant detection scenarios, and plays a positive role in the prevention and control of Mycoplasma pneumoniae infection. It has broad application prospects and important clinical value. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below according to specific embodiments of the present invention in conjunction with the accompanying drawings, wherein

[0032] Figure 1 The structure diagram of the colloidal gold test strip for Mycoplasma pneumoniae in Example 1 of the present invention, wherein 1 is a sample pad, 2 is a binding pad, 3 is a nitrocellulose membrane, 31 is a detection line, 32 is a quality control line, 4 is a water absorbent pad, and 5 is a base plate. DETAILED DESCRIPTION

[0033] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0034] Some of the reagents involved in the following examples are as follows:

[0035] The sample pad treatment solution consists of 50 mM Tris-HCl, 0.6 wt % casein, 0.8 wt % polyvinylpyrrolidone K30, 1 wt % surfactant S9, 0.05 wt % Tween 20, 0.05 wt % ProClin 300 and 0.004 wt % heterophilic antibody blocker HBR, and the solution pH = 7.5.

[0036] The conjugate pad treatment solution consisted of 50 mM Tris-HCl, 0.5 wt % BSA, 1 wt % polyvinylpyrrolidone K30, 0.05 wt % Tween 20 and 0.05 wt % ProClin 300, and the solution pH = 8.5.

[0037] The colloidal gold solution has an OD value of 8-10 and is prepared by heating and calcining 0.04 wt % of chloroauric acid and 0.06 wt % of sodium citrate.

[0038] The colloidal gold labeling complex solution consists of 50 mM Tris-HCl, 0.6 wt % casein, 15 wt % sucrose, 5 wt % trehalose and 0.05 wt % ProClin 300, and the solution pH=8.

[0039] The coating diluent consisted of 10 mM PBS and 1.5 wt % sucrose, and the solution pH was 7-7.5.

[0040] Example 1: Preparation of test strips

[0041] 1. Pre-treat the sample pad and conjugate pad

[0042] (1) Sample pad pretreatment: 0.08 mL / cm 2 The sample pad is immersed in the sample pad treatment solution, and dehydrated after 5 minutes. The dehydrated sample pad is dried at 45°C for 7 hours and sealed in a low humidity environment.

[0043] (2) Pretreatment of the conjugate pad: 0.06 mL / cm 2 The conjugate pad is immersed in the conjugate pad treatment solution and dehydrated after 5 minutes. The dehydrated conjugate pad is dried at 45°C for 7 hours and sealed in a low humidity environment.

[0044] 2. Colloidal gold labeling and reconstitution of detection antibodies

[0045] (1) Labeling and reconstitution of mouse anti-human Mycoplasma pneumoniae detection antibodies:

[0046] Weigh 1 mL of colloidal gold solution and adjust the pH value of the colloidal gold solution to 7.2-7.5 using 0.1 M potassium carbonate solution;

[0047] Add mouse anti-human Mycoplasma pneumoniae detection antibody to the adjusted colloidal gold solution to make the final concentration 15 μg / mL, and shake for more than 10 minutes to fully mix;

[0048] After mixing, add bovine serum albumin BSA to a final concentration of 1 wt %, and shake for more than 10 minutes to fully mix;

[0049] The mixed solution was subjected to low-temperature high-speed centrifugation at 4°C and 10,000 rpm for 8 minutes;

[0050] The supernatant was removed, and a colloidal gold labeled reconstitution solution was added to the precipitate, and the precipitate was broken up and resuspended to make the OD value of the solution between 120 and 180, thereby obtaining a mouse anti-human Mycoplasma pneumoniae detection antibody colloidal gold labeled reconstitution solution.

[0051] (2) Labeling and reconstitution of chicken IgY antibody:

[0052] Weigh 1 mL of colloidal gold solution and adjust the pH value of the colloidal gold solution to 8.5-9.0 using 0.1 M potassium carbonate solution;

[0053] Add chicken IgY antibody to the adjusted colloidal gold solution to a final concentration of 10 μg / mL, and shake for more than 10 minutes to fully mix;

[0054] After mixing, add bovine serum albumin BSA to a final concentration of 1 wt %, and shake for more than 10 minutes to fully mix;

[0055] The mixed solution was subjected to low-temperature high-speed centrifugation at 4°C and 10,000 rpm for 8 minutes;

[0056] The supernatant was removed, and a colloidal gold labeled reconstitution solution was added to the precipitate to break up and resuspend the precipitate so that the OD value of the solution was between 150 and 200, thereby obtaining a chicken IgY antibody colloidal gold labeled reconstitution solution.

[0057] 3. Capture Antibody Coating

[0058] (1) Coating of mouse anti-human Mycoplasma pneumoniae capture antibody: dilute the mouse anti-human Mycoplasma pneumoniae antibody to 0.8 mg / mL using a coating diluent to obtain a mouse anti-human Mycoplasma pneumoniae capture antibody coating solution;

[0059] (2) Coating of goat anti-chicken IgY antibody: Use coating diluent to dilute goat anti-chicken IgY antibody to 1 mg / mL to obtain goat anti-chicken IgY antibody coating solution.

[0060] 4. Assemble the test strips

[0061] The mouse anti-human pneumoniae Mycoplasma detection antibody colloidal gold labeled re-solution prepared above and the chicken IgY antibody colloidal gold labeled re-solution prepared above are mixed evenly, and the concentration of the mouse anti-human pneumoniae Mycoplasma detection antibody colloidal gold labeled re-solution is adjusted to OD60, and the concentration of the chicken IgY antibody colloidal gold labeled re-solution is adjusted to OD25, and the re-solutions are sprayed on the treated conjugate pad at a spray volume of 3 μL / cm, and the sprayed conjugate pad is placed at a temperature of 45° C. and baked for 6 hours, and then sealed and stored in an environment with a humidity of ≤30%;

[0062] The mouse anti-human pneumoniae Mycoplasma capture antibody coating solution and the goat anti-chicken IgY antibody coating solution were sprayed on the nitrocellulose membrane at an amount of 1 μL / cm to form a detection line and a quality control line. The treated nitrocellulose membrane was baked at 45°C for 8 hours and sealed and stored in an environment with a humidity of ≤30%.

[0063] The sample pad, conjugate pad, absorbent pad, and nitrocellulose membrane are all bonded to the bottom plate, wherein the nitrocellulose membrane is located near the middle of the bottom plate, one end of the nitrocellulose membrane is bonded to the absorbent pad with a 2mm overlap, and the other end is bonded to the conjugate pad with a 2mm overlap, and the other end of the conjugate pad is bonded to the sample pad with a 2mm overlap. The bottom plate is located at the bottom to support the entire test strip. The test strip structure is as follows: Figure 1 shown.

[0064] Example 2: Use of test strips

[0065] (1) Use sample diluent to dilute the sample to a concentration of 10 7 CFU / mL of Mycoplasma pneumoniae antigen standard was diluted to the following concentrations: 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 CFU / mL;

[0066] (2) Take 80 μL of the four different concentrations of Mycoplasma pneumoniae antigen standard solutions prepared in step (1) and add them dropwise to the sample pad of the test strip prepared in Example 1. Due to capillary action, the sample will move along the test strip to the conjugate pad and nitrocellulose membrane. The result will be observed after 10 minutes. After 15 minutes, the color development will be invalid.

[0067] (3) Mycoplasma pneumoniae antigen concentration ≥ 10 3 The CFU / mL test result is positive, less than 10 3 The CFU / mL test result was negative.

[0068] Comparative Example 1: Adjusting the detection antibody and capture antibody loading

[0069] (1) A test strip was prepared according to the method described in Example 1, except that when the colloidal gold-labeled mouse anti-human Mycoplasma pneumoniae detection antibody reconstitution solution and the colloidal gold-labeled chicken IgY antibody reconstitution solution were mixed, the concentration of the colloidal gold-labeled mouse anti-human Mycoplasma pneumoniae detection antibody was adjusted to OD30;

[0070] According to the method described in Example 2, the concentration of 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 The minimum detection limit was 10 4 CFU / mL;

[0071] (2) A test strip was prepared according to the method described in Example 1, except that: when preparing the antibody coating solution, the concentration of the mouse anti-human Mycoplasma pneumoniae capture antibody was diluted to 0.5 mg / mL;

[0072] According to the method described in Example 2, the concentration of 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 The minimum detection limit was 10 4 CFU / mL

[0073] From the above results, it can be seen that reducing the amount of detection antibody on the conjugate pad or the amount of capture antibody on the nitrocellulose membrane will affect the detection performance of the reagent strip.

[0074] Comparative Example 2: Adjusting the pH value of the sample pad treatment solution

[0075] (1) preparing a test strip according to the method described in Example 1, except that the pH of the sample pad treatment solution was adjusted to 8.5-9;

[0076] According to the method described in Example 2, the concentration of 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 The minimum detection limit was 10 4 CFU / mL.

[0077] (2) A test strip was prepared according to the method described in Example 1, except that the pH of the sample pad treatment solution was adjusted to 6-6.5;

[0078] According to the method described in Example 2, the concentrations of 0 CFU / mL, 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 The test was performed with a Mycoplasma pneumoniae antigen standard of 0 CFU / mL, and the result showed that the Mycoplasma pneumoniae antigen standard of 0 CFU / mL was positive, indicating that the test strip had a false positive.

[0079] From the above results, it can be seen that the pH value of the sample pad treatment solution will affect the detection performance of the test strip. A too high pH value will reduce the sensitivity of the test strip, and a too low pH value will easily cause false positives in the test strip.

[0080] Comparative Example 3: Comparison of detection effect with commercially available test strips

[0081] Take the test strip prepared in Example 1 and the commercially available test strip of the control example, and test the concentration of 10 2 CFU / mL, 10 3 CFU / mL, 10 4 CFU / mL, 10 5 The results are shown in Table 1.

[0082] Table 1 Comparison of different concentrations of Mycoplasma pneumoniae antigen standards

[0083]

[0084] From the above results, it can be seen that the Mycoplasma pneumoniae test strip provided by the present invention has a low Mycoplasma pneumoniae antigen level of 10 3 CFU / mL, it can still detect positive results, and the detection sensitivity is significantly improved compared with commercially available test strips.

[0085] Obviously, the above embodiments are merely examples for clear explanation and are not intended to limit the implementation methods. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation methods here. The obvious changes or modifications derived from these are still within the protection scope of the invention.

Claims

1. A method for preparing a colloidal gold test strip for Mycoplasma pneumoniae, characterized in that: The preparation method specifically comprises the following steps: (1) Soaking the sample pad in a sample pad treatment solution, wherein the sample pad treatment solution comprises a buffer, polyvinyl pyrrolidone, a surfactant, a preservative and a blocking agent, the pH value of the sample pad treatment solution is 7-8, and the amount of the sample pad treatment solution added is 0.05-0.1 mL / cm 2 ; (2) taking a colloidal gold solution, adding a Mycoplasma pneumoniae detection antibody and bovine serum albumin to make the final concentration of the Mycoplasma pneumoniae detection antibody be 13-18 μg / mL, centrifuging to obtain a precipitate, adding a colloidal gold marker reconstitution solution to the precipitate to obtain a colloidal gold-labeled Mycoplasma pneumoniae detection antibody reconstitution solution, wherein the concentration of the colloidal gold solution is OD8-10; Take the colloidal gold solution, add the quality control detection antibody and bovine serum albumin to make the final concentration of the quality control detection antibody be 8-12 μg / mL, centrifuge to obtain the precipitate, add the colloidal gold marker re-solution to the precipitate, and obtain the colloidal gold-labeled quality control detection antibody re-solution; The colloidal gold marker complex solution includes Tris-HCL, casein or bovine serum albumin, sucrose, trehalose, sodium azide or ProClin300; The colloidal gold-labeled Mycoplasma pneumoniae detection antibody reconstitution solution and the colloidal gold-labeled quality control detection antibody reconstitution solution are evenly mixed to obtain a mixed reconstitution solution, and the mixed reconstitution solution is sprayed on the conjugate pad; (3) Setting a test line and a quality control line on the coating membrane, and spraying a solution containing Mycoplasma pneumoniae capture antibody and a solution containing quality control capture antibody respectively; The final concentration of the Mycoplasma pneumoniae capture antibody is 0.6-1 mg / mL, and the final concentration of the quality control capture antibody is 0.8-1.2 mg / mL; (4) Assembling the sample pad, the binding pad, the coating membrane and the water-absorbing pad to obtain the Mycoplasma pneumoniae colloidal gold test strip.

2. The preparation method according to claim 1, characterized in that: The method further includes a step of pretreating the conjugate pad, wherein the pretreatment is to immerse the conjugate pad in a conjugate pad treatment solution, wherein the conjugate pad treatment solution is composed of 40-80 mM Tris-HCl, 0.4-0.8 wt % casein or bovine serum albumin, 0.8-1.5 wt % polyvinyl pyrrolidone K30, 0.04-0.08 wt % Tween 20, and 0.02-0.06 wt % sodium azide or ProClin300.

3. The preparation method according to claim 2, characterized in that: The amount of the binding pad treatment solution added is 0.04-0.08 mL / cm 2 .

4. The preparation method according to claim 1, characterized in that: In step (1), the sample pad treatment solution consists of 40-80 mM Tris-HCl, 0.4-0.8 wt % casein or BSA, 0.8-1.5 wt % polyvinylpyrrolidone K30, 0.8-1.5 wt % surfactant S9, 0.04-0.1 wt % Tween 20, 0.02-0.05 wt % sodium azide or ProClin300 and 0.002-0.006 wt % heterophilic antibody blocker HBR.

5. The preparation method according to claim 1, characterized in that: In step (2), the concentration of the colloidal gold-labeled Mycoplasma pneumoniae detection antibody in the mixed reconstitution solution is OD45-70, and the concentration of the colloidal gold-labeled quality control detection antibody is OD20-30.

6. The preparation method according to claim 1, characterized in that: In step (2), the spraying amount of the mixed solution is 2.5-4 μL / cm.

7. The preparation method according to claim 1, characterized in that: In step (3), the spraying amount of the solution containing the Mycoplasma pneumoniae capture antibody on the detection line is 1-2 μL / cm.

8. A colloidal gold test strip for Mycoplasma pneumoniae, characterized in that: It is prepared by any method of claims 1-7.

9. The Mycoplasma pneumoniae colloidal gold test strip according to claim 8, characterized in that: The Mycoplasma pneumoniae colloidal gold test strip also includes a sample diluent.

10. Use of the colloidal gold test strip according to claim 8 or 9 in preparing a pneumonia auxiliary diagnosis product.

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