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Immune chromatography test paper for inspecting legionella pneumophilia antibody and its production

A technology of immunochromatographic test paper and Legionella pneumophila, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of difficult standardization of experimental systems and cumbersome operation steps, and achieve easy industrial production, broad application prospects, and detection The effect of simple method

Inactive Publication Date: 2007-11-21
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But in practice, the use of these methods has exposed some common disadvantages, such as: the experiment must be carried out in a special laboratory; power and special instruments are required; operators must be specially trained; the experimental system is not easy to standardize. The results are credible, and it is necessary to conduct multiple pre-tests and set up multiple controls; the test operation steps are cumbersome and need at least 2 hours

Method used

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  • Immune chromatography test paper for inspecting legionella pneumophilia antibody and its production

Examples

Experimental program
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Effect test

Embodiment 1

[0034] Example 1. Preparation of immunochromatographic test paper for detecting Legionella pneumophila antibodies

[0035] 1. Preparation of Legionella pneumophila antigen

[0036] The Legionella pneumophila serotype 1 strain (purchased from the strain bank of the Institute of Microbiology and Epidemiology of the Academy of Military Medical Sciences, deposit number 510101) was inoculated on BCYE plates (the formula is CM655 basic medium, SR110A growth additives, SR118E selective additives) , At 37℃, 5% CO 2 Incubate in an incubator, observe the morphology of the colonies on the plate, pick a typical single colony, and place it on a BCYE slope at 37℃, 5% CO 2 Incubate in an incubator for 3-5 days; wash the cultivated slope with sterile PBS (0.01M pH 7.2) and wash the lawn, centrifuge at 8000 rpm for 25 minutes, discard the supernatant, and collect the bacterial pellet; each gram of wet weight is suspended in In 10mL 0.1M NaOH, stir at room temperature for 1 hour; adjust the pH of t...

Embodiment 2

[0055] Example 2. Detection of Legionella pneumophila antibodies and cross-tests with other related bacterial serum samples

[0056] 1. Detection of antibodies to different serotypes of Legionella pneumophila

[0057] Rabbits were immunized with antigens of types 1, 2, 3, 4, 5, 6, 7, 9, and 10 of Legionella pneumophila from the Microbiological Testing and Research Center of the Institute of Microbiology and Epidemiology of the Academy of Military Medical Sciences (immunization dose is 2 mg / mL / mouse, the number of immunizations is 4-6, and each immunization interval is 10 days) The obtained antiserum sample is diluted with normal saline at a ratio of 1:40 and used as a sample test solution for use.

[0058] Take the kit prepared in Example 1 containing the immunochromatographic test paper for detecting Legionella pneumophila antibodies of the present invention, add 3 drops (about 150ul) of the sample detection solution to the spotting port respectively, and start to observe the re...

Embodiment 3

[0065] Example 3: Laboratory assessment

[0066] 1. Sample preparation

[0067] The Legionella pneumophila type 1-10, Escherichia coli, Salmonella, Shigella dysentery, Pseudomonas aeruginosa, Yersinia pestis, B. The antiserum samples of Lutella and Staphylococcus aureus were diluted with normal saline at a ratio of 1:40 and used as the sample detection solution for later use.

[0068] 2. Experimental method

[0069] Take the kit prepared in Example 1 containing the immunochromatographic test paper for detecting Legionella pneumophila antibodies of the present invention, add 3 drops (about 150ul) of the above sample detection solution to the spotting port respectively, and start to observe the results after 2 minutes, 15 Minute observation is terminated.

[0070] Result report: Only one red precipitation line appeared in the quality control observation window "C" (quality control line), which is negative, that is, no Legionella pneumophila antibody was detected; in the detection ob...

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Abstract

An immune chromatographic test paper used for detecting antibody of legionella pneumophilia consists of sample pad, gold label pad containing A label colloidal gold probe of staphylococcus aureus protein, nitric acid fiber membrane and water absorption pad. It is featured as enveloping detection line containing antigen and quality control line containing antibody on nitric acid fiber membrane and setting said detection line to be separated from said quality control line.

Description

Technical field [0001] The invention relates to a test paper for detecting Legionella pneumophila antibodies and a preparation method thereof, in particular to an immunochromatographic test paper for detecting Legionella pneumophila antibodies and a preparation method thereof. Background technique [0002] Legionella (Legionella) was discovered in 1976. At that time, American veterans gathered in Philadelphia, during which a severe pneumonia of unknown cause broke out. 149 participants were infected and 34 died. A new strain was isolated from the lung tissue of the deceased and named Legionella. At present, 43 species and 65 serotypes of Legionella have been discovered. Among them, Legionella pneumophila (LP) is most closely related to human diseases. Studies have shown that 80%-85% of Legionella pneumonia Caused by LP, 15 serotypes of LP have been found. [0003] Legionnaires' disease (Legionnarires' disease, LD) is an acute bacterial respiratory infection caused by Legionella i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/532
Inventor 端青张宝元汪黎朱虹檀华何君左婷庭宋立华
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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