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Immunological chromatographic test paper for testing francisella tularensis and its preparation method

An immunochromatographic test strip, Tularemia technology, which is applied in the direction of material analysis, measuring device, analysis material, etc. by observing the effect on chemical indicators, can solve problems such as unsuitability for use and long-term sample processing, and achieve sample Handling simple effects

Inactive Publication Date: 2011-02-16
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although immunofluorescence test and PCR have been well applied in the laboratory, these two tests require special instruments and reagents, must be operated by experienced technicians in the laboratory, and the specimen processing takes a long time , so it is not suitable for use at the grassroots level and emergency scene

Method used

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  • Immunological chromatographic test paper for testing francisella tularensis and its preparation method
  • Immunological chromatographic test paper for testing francisella tularensis and its preparation method
  • Immunological chromatographic test paper for testing francisella tularensis and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1, the preparation of the immunochromatography test paper that detects Francisella tularensis

[0038] 1. Preparation of Francisella tularensis monoclonal antibody

[0039] 1) Preparation of Francisella tularensis monoclonal antibody

[0040] BALB / c mice were used as immunized animals, and multiple injections of Freund's complete adjuvant Francisella tularensis vaccine strain (Institute of Microbial Epidemiology, Academy of Military Medical Sciences, preservation number 410062) immunogen emulsifier were injected subcutaneously on the back of the neck at intervals of 2- Take the same dose of immunogen plus the same amount of Freund's incomplete adjuvant mixed emulsification in 3 weeks, boost once for booster immunization, and once for booster immunization in the abdominal cavity after the fourth immunization, take spleen cells 3 days later to fuse with SP2 / 0-Ag14 myeloma cells, use the following The formaldehyde-fixed antigen of Francisella tularensis whole b...

Embodiment 2

[0063] Embodiment 2, the detection of Francisella tularensis and the cross test with other relevant bacteria

[0064] 1. Detection of Francisella tularensis

[0065] 1) Five strains from different sources were purchased from the Microbial Testing Research Center of the Institute of Microbial Epidemiology, Academy of Military Medical Sciences, at a concentration of 1×10 5 Francisella tularensis strains in cfu / ml: Francisella tularensis 410101, Francisella tularensis 410102, Francisella tularensis 410103, Francisella tularensis 410104, Francisella tularensis 410105, bacterial suspension as a sample The detection solution is ready for use.

[0066] 2) The monoclonal antibody secreted by the coated tularemia hybridoma cell line Ft5CGMCC No.1566 prepared in Example 1 and the immunochromatographic test paper of goat anti-mouse IgG were all positive results.

[0067] 2. Cross-test with other related bacteria

[0068] 1) Bought from the Microbial Testing and Research Center of the ...

Embodiment 3

[0070] Example 3. Detection of clinical and environmental suspected Francisella tularensis specimens

[0071] During the measurement, add 3 drops (150ul) of the above-mentioned sample detection solution to the sample holes of the kit prepared in Example 1, and start to observe the results through the detection window after 2 minutes, and the observation is terminated after 15 minutes. Result report: one red precipitation line at the quality control line is negative, and there is no precipitation line at the detection line, that is, no Francisella tularensis is detected; one precipitation line at the quality control line and one detection line is positive, That is, Francisella tularensis was detected. There is no red precipitation line at the quality control line and test line, indicating that the test paper is invalid.

[0072] 1) Purely cultured bacteria: Pick a single colony of suspected tularemia, and make 0.5ml of normal saline bacterial suspension in a small test tube as...

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Abstract

An immune chromatographic test paper for testing francisella tularensis is prepared as series-connecting sample pad, golden labeled pad with collaurum probe containing francisella tularensis single clone antibody label, fiber membrane nitrate and water absorption pad; enveloping a detection line and a quality control line being separated to each other on fiber membrane nitrate; using francisella tularensis single clone antibody as detection line and using anti-antibody of anti-mouse as quality control line.

Description

technical field [0001] The invention relates to a test paper for detecting Francisella tularensis and a preparation method thereof, in particular to an immunochromatographic test paper for detecting Francisella tularensis and a preparation method thereof. Background technique [0002] Francisella tularensis is an important biological warfare agent, which has been included in the checklist of pathogenic microorganisms in the International Convention on the Prohibition of Biological Weapons. In June 2001, the American Journal of Microbiology published a "The Working Group on Civilian Biodefense" report on the risk assessment of Francisella tularensis as a biological weapon for terrorist attacks. This working group is composed of 25 experts from American medical scientific research institutions, the military, and relevant government departments. After searching the relevant literature from January 1966 to October 2000, it is finally concluded that Francisella tularensis has bee...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558G01N33/532G01N21/78
Inventor 端青檀华朱虹何君左庭婷苏裕心
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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