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Immunization chromatograph test paper for detecting plague yersinia and preparation method thereof

An immunochromatographic test strip, Yersinia technology, applied in biological testing, material analysis by observing the effect on chemical indicators, measuring devices, etc. Simple effect of specimen handling

Inactive Publication Date: 2010-05-05
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sensitivity limitations may be due to assay conditions, PCR inhibitors in the template, and small sample sizes for DNA extraction
Therefore, this method cannot be used as a routine diagnostic method for plague (Rahalison L, Vololonirina E, Ratsitorahina M.Diagnosis of bubonic plague by PCR in Madagascar underfield conditions J Clin Microbiol.2000Jan; 38(1):260-3.)

Method used

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  • Immunization chromatograph test paper for detecting plague yersinia and preparation method thereof
  • Immunization chromatograph test paper for detecting plague yersinia and preparation method thereof
  • Immunization chromatograph test paper for detecting plague yersinia and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1, the preparation of the immunochromatographic test paper that detects Yersinia pestis

[0037] 1. Preparation of antibodies against Yersinia pestis F1 antigen

[0038] Healthy large-eared white rabbits with a body weight of 2 kg were selected and injected subcutaneously with Freund’s complete adjuvant Yersinia pestis F1 antigen 2 mg / rat (purchased from Changchun Institute of Biological Products), and respectively on the 20th day after the initial immunization, On the 30th and 40th days, an additional shot of immunization solution was added. The dose and route were the same as the initial immunization. The antibody titer in the serum was tested 10 days after the last immunization, and the antibody titer was detected by the agar two-way diffusion method to reach 1:32 or more. Blood collection .

[0039] The anti-Yersinia pestis F1 antigen antibody in the blood was purified by saturated ammonium sulfate salting-out method, and the antibody obtained by agar tw...

Embodiment 2

[0053] Embodiment 2, the detection of Yersinia pestis and its F1 antigen and the cross test with other bacteria

[0054] 1. Detection of Yersinia pestis F1 antigen

[0055] 1) The F1 antigen of Yersinia pestis (purchased from Changchun Institute of Biological Products) was mixed with normal saline from 1000 μg / mL to 100, 10, 1, 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 After the μg / mL gradient dilution, it was used as the sample detection solution for later use.

[0056] 2) Take the immunochromatography kit for detecting Yersinia pestis prepared in step 5 of Example 1, add 3 drops (about 150 μL) of the sample detection solution in step 1) to the sample wells, and start to observe the results after 5 minutes , observe for 15min. The results show that the concentrations are 100, 10, 1, 10 -1 、10 -2 、10 -3 For the sample detection solution of μg / mL, a red precipitation line appeared on both the test strip and the control strip, and the test result was positive.

[0057]...

Embodiment 3

[0061] Embodiment 3, Different batches of Yersinia pestis immunochromatographic kits are effective against Yersinia pestis, Yersinia pestis F 1 Antigen detection and cross test with other bacteria

[0062] 1. Different batches of Yersinia pestis immunochromatographic kits against Yersinia pestis, Yersinia pestis F 1 antigen detection

[0063] Prepare three batches of Yersinia pestis immunochromatography kits according to the method of Example 1, the batch numbers are 200401, 200402 and 200403 respectively, and use it to detect liquids of Yersinia pestis EV strains and Evp strain samples of different concentrations and Yersinia pestis F 1 Antigen is detected, and detection result is as shown in table 1, table 2 and table 3, shows that application Yersinia pestis immunochromatography kit of the present invention detects different Yersinia pestis strains, and minimum detection unit is 156,000 CFU / mL, detection of Yersinia pestis F 1Antigen, the minimum detection unit is 1ng / m...

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Abstract

A method for preparing immune chromatographic test paper used to detect Yersinia pestis includes setting water absorption paper pad, setting cellulose membrance closely to said paper pad, setting goldstandard pad of immune colloidal gold probe containing F1 antigen - antibody resisting Yersinia pestis closely to cellulose membrance and setting sample pad closely to said gold standard pad, settinga test belt containing said F1 antigen - antibody and a comparison belt containing antiantibody of said antigen - antibody on cellulose membrane.

Description

technical field [0001] The invention relates to a test paper for detecting pathogenic bacteria and a preparation method thereof, in particular to an immunochromatographic test paper for detecting Yersinia pestis and a preparation method thereof. Background technique [0002] Plague, also known as the Black Death, is an infectious disease with a high transmissibility and fatality rate. Human plague is divided into three types according to its onset: bubonic plague, pneumonic plague and septicemic plague. Among them, the incubation periods of bubonic plague and pneumonic plague are 2-10 days and 2-3 days respectively, and the case fatality rates are as high as 50% and 100% respectively. Therefore, monitoring the epidemic and early diagnosis of plague is an important part of plague prevention and control. [0003] At present, more than 20 countries in the world have reports of plague outbreaks in local areas every year. Although it has been more than a century since the disco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/547G01N33/52G01N21/78G01N33/533
Inventor 端青檀华何君朱虹苏裕心
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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