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Rv1793 recombinant protein for serodiagnosis of medicament-resistant tuberculosis

A technology of rv1793 and recombinant protein, which is applied in the field of recombinant protein in serological diagnosis, can solve the problems of strong sensitivity and no specificity found

Inactive Publication Date: 2011-08-24
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, each antigen can only detect a part of the infected patients, and so far no diagnostic antigen with high specificity and sensitivity has been found for drug-resistant tuberculosis

Method used

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  • Rv1793 recombinant protein for serodiagnosis of medicament-resistant tuberculosis
  • Rv1793 recombinant protein for serodiagnosis of medicament-resistant tuberculosis
  • Rv1793 recombinant protein for serodiagnosis of medicament-resistant tuberculosis

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Experimental program
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Effect test

Embodiment

[0028] Coat 100 μL / well of active recombinant Rv1793 protein at a concentration of 5 μg / mL at 4° overnight; 300 μL / well in PBST, 3 min each time, wash 3 times, and pat dry; use 1% BSA in PBS, 300 μL / well, Incubate at 37°C for 2 hours to block; 300 μL / well PBST, 3 minutes each time, wash 3 times, pat dry; add 1:100 diluted healthy serum and patient serum to be tested, 100 μL / well, incubate at 37°C for 1 hour; 300 μL / well PBST, 5 min each time, wash 5 times, pat dry; add freshly diluted enzyme-labeled antibody 300ul to each reaction well, incubate at 37°C for 1 h; 300μL / well PBST, 5 min each time, wash 5 times, pat dry; Add 100 μL of TMB chromogenic solution to each reaction well, and incubate at 37°C for 5 min; add 50 μL of stop solution to each reaction well, measure the OD value of each well after zeroing the blank control well on a microplate reader at 450 nm. Then take the mean value of the healthy control serum plus 2 times the standard deviation as the standard, if it is ...

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Abstract

The invention belongs to the technical field of tuberculosis diagnosis, and particularly discloses a recombinant protein for serodiagnosis of multimedicament-resistant tubercle bacilli. The recombinant protein is Rv1793 recombinant protein, and is obtained by the following steps of: inserting Rv1793 into the position between Pet28a EcoRI and HindIII endonuclease sites to form a recombinant plasmid, transforming the recombinant plasmid into E. coli BL21 to form recombinant E. coli, and expressing the protein. The sensitivity of the novel recombinant protein on the serodiagnosis of medicament resistance can reach 75 percent, and is far more than 53.3 percent of diagnosis rate of medicament resistance of a control CFP-10 recombinant protein. The Rv1793 recombinant protein becomes an effective biomarker applied to the serodiagnosis of medicament-resistant tuberculosis.

Description

technical field [0001] The invention belongs to the technical field of serological diagnosis of tuberculosis, and in particular relates to a recombinant protein used for the serological diagnosis of multidrug-resistant Mycobacterium tuberculosis. Background technique [0002] The first-line anti-tuberculosis drugs commonly used in clinic include isoniazid (INH), rifampin (RFP), pyrazinamide (PZA), streptomycin (SM) and B Drug resistance to ethambutol (EMB) exists, even to multiple drugs at the same time, that is, MDR-TB appears. MDR-TB refers to the resistance of Mycobacterium tuberculosis expelled from tuberculosis patients to at least two or more drugs including INH and RFP. According to WHO estimates, 50 million people worldwide are infected with drug-resistant tuberculosis, of which China accounts for the most, accounting for 11%. The outbreak of drug-resistant or even multidrug-resistant tuberculosis has become a serious public health problem worldwide. [0003] Since...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/35C12N15/31C12N15/70G01N33/569
Inventor 王洪海彭超张鹭刘丹徐文玺何磊王文婕郝科威
Owner FUDAN UNIV
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