Immunochromatographic test paper for detecting mycobacterium bovis antibodies and preparation method thereof

A technology of mycobacterium tuberculosis and immunochromatographic test paper, which is applied in the direction of analysis materials, measuring devices, instruments, etc., to achieve the effect of simple detection method, low cost, and easy industrial production

Inactive Publication Date: 2010-04-07
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

[0009] At present, the use of colloidal gold immunochromatography for in vitro detection of Mycobacterium bovis is still blank in China

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] The preparation of embodiment 1, Mycobacterium tuberculosis bovis MPB70, MPB83, ESXA and ESXB

[0043] 1. Use the PCR method to amplify the MPB70 full DNA, and its primers are:

[0044] 1: 5'-CGCGGATCCATGAAGGTAAAGAACACAATTGC-3';

[0045] 2: 5'-CCCAAGCTTTTACGGAGGCATTAGCACGCTGT-3'.

[0046] The PCR method is used to amplify the MPB83 full DNA, and its primers are:

[0047] 1: 5'-CGCGGATCCATGATCAACGTTCAGGCAAACC-3';

[0048] 2: 5'-CCCAAGCTTTTACAGCACCGTATCGATCATG-3'.

[0049] Use the PCR method to amplify the whole ESXA DNA, and the primers are:

[0050] 1: 5'-CGCGGATCCATGACCATCAACTATCAATTCGG-3';

[0051] 2: 5'-CCCAAGCTTTTAGGCCCAGCTGGAGCCGAC-3'.

[0052] Use the PCR method to amplify the whole ESXB DNA, and the primers are:

[0053] 1: 5'-GGATCCATGGCAGAGATGAAGACCG-3';

[0054] 2: 5'-AAGCTTTCAGAAGCCCATTTGCGAGG-3'.

[0055] The above primers all introduce BamHI and HindIII restriction enzyme sequences

[0056] 2. Cut out the target fragment from the T vector, connect...

Embodiment 2

[0089] Embodiment 2, the detection of Mycobacterium tuberculosis bovis antibody and the cross test with other relevant bacterial serum samples

[0090] 1. Crossover test of other related bacterial serum samples

[0091] Escherichia coli (preservation number 270014), Salmonella (preservation number 460046), Shigella dysenteriae (preservation number 51258), and Pseudomonas aeruginosa purchased from the Microbial Testing Research Center of the Institute of Microbial Epidemiology, Academy of Military Medical Sciences Bacteria (preservation number 10101), Yersinia pestis (preservation number 410050), Brucella (preservation number 55227), Staphylococcus aureus (preservation number 26067) antiserum samples were used as sample detection solution for standby use, and bovine tuberculosis branch Bacillus antiserum was used as control.

[0092] Result report: Serum titer above 1:40 is the positive diagnostic standard, and only one red precipitation line (control) appears at the quality c...

Embodiment 3

[0094] Embodiment 3, laboratory assessment

[0095] 1. Sample preparation

[0096] Legionella pneumophila type 1-10, Escherichia coli, Salmonella, Shigella dysenteriae, Pseudomonas aeruginosa, Yersinia pestis, cloth The antiserum samples of Rutella and Staphylococcus aureus were diluted with normal saline at a ratio of 1:40 and then used as sample detection solution for later use.

[0097] 2. Experimental method

[0098] Get the test kit that the immunochromatographic test paper that the present invention detects Mycobacterium bovis antibody that the embodiment 1 prepares is housed, add above-mentioned sample detection solution 3 drops (about 150ul) respectively in the sampling port, begin to observe the result after 2 minutes , 15 minutes to observe the termination.

[0099] Result report: Only one red precipitation line at the quality control observation window "C" (quality control line) is negative, that is, no Mycobacterium bovis antibody is detected; at the detection o...

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Abstract

The invention discloses immunochromatographic test paper for detecting mycobacterium bovis antibodies and a preparation method thereof. The test paper comprises a sample pad, a conjugate pad closely which is connected with a labeling colloidal gold probe and contains staphylococcus aureus protein A at one end of the sample pad, a nitrocellulose membrane closely connected with the other end of the conjugate pad and a absorbent pad closely connected with the other end of the nitrocellulose membrane, wherein the nitrocellulose membrane is coated with a detection line and a control line which are separated with each other; the detection line comprises mycobacterium bovis antigens; and the control line comprises antibodies capable of specifically bound with the staphylococcus aureus protein A. The immunochromatographic test paper has the advantages of simplicity, sensitivity, specificity and high speed and is suitable for clinical and field use.

Description

technical field [0001] The invention relates to a test paper for detecting bovine mycobacterium tuberculosis antibody and a preparation method thereof, in particular to an immunochromatographic test paper for detecting bovine mycobacterium tuberculosis antibody and a preparation method thereof. Background technique [0002] Since Kock discovered Mycobacterium tuberculosis in 1882, more than 100 species of mycobacteria have been reported. There are many classifications of mycobacteria. Internationally, mycobacteria are generally divided into typical tuberculosis mycobacterium (TM) and non-tuberculosis mycobacterium (NTM). TM includes Mycobacterium tuberculosis human, Mycobacterium bovis, Mycobacterium africanus and Mycobacterium microti, also known as Mycobacterium tuberculosis complex (MTBC), the DNA related About 85% to 100%, with similar taxonomic status; NTM refers to all other mycobacteria except MTBC and Mycobacterium leprae. [0003] Tuberculosis is a zoonotic infect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/532
Inventor 端青赵海朱虹何君檀华李岩伟
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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