Combined protein and detection reagent for the detection of Mycobacterium tuberculosis
A technology of mycobacterium tuberculosis and combined proteins, which is applied in the field of medical immunological detection, can solve the problems of delayed clinical diagnosis and treatment, long detection cycle, high price, etc., and achieve the effect of improving the cure rate, accurate diagnosis and treatment, and avoiding transmission
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Embodiment 1
[0039] The preparation method of the detection reagent for Mycobacterium tuberculosis is as follows:
[0040] 1), the synthetic gene fragments of the first combined protein of the present invention and the second combined protein are excised from the cloning vector with restriction endonucleases NdeI and XhoI, cloned into the expression vector PET28b, in Escherichia coli BL21 (DE3) After cultured to OD600 of 0.6, IPTG was added to the final concentration of 0.1M to induce expression, and the identification and analysis were carried out by gel electrophoresis and Western Blotting;
[0041] 2), adopt gel (Pharmacia company chelating sephrose fast flow production), purify expression protein through metal chelate chromatography;
[0042] 3), immunize rabbits with human IgG (purchased from Tiangen Biotechnology Co., Ltd.) using standard immunization procedures to prepare rabbit immunized human IgG antibodies (also including IgM, IgA);
[0043] 4), prepare colloidal gold by citric ...
Embodiment 2
[0053] The preparation method of the detection reagent for Mycobacterium tuberculosis is as follows:
[0054] 1), the synthetic gene fragments of the first combined protein of the present invention and the second combined protein are excised from the cloning vector with restriction endonucleases NdeI and XhoI, cloned into the expression vector PET28b, in Escherichia coli BL21 (DE3) After cultured to OD600 of 0.6, IPTG was added to the final concentration of 0.1M to induce expression, and the identification and analysis were carried out by gel electrophoresis and Western Blotting;
[0055] 2), adopt gel (Pharmacia company chelating sephrose fast flow production), purify expression protein through metal chelate chromatography;
[0056] 3), immunize rabbits with human IgG (purchased from Tiangen Biotechnology Co., Ltd.) using standard immunization procedures to prepare rabbit immunized human IgG antibodies (also including IgM, IgA);
[0057] 4), prepare colloidal gold by citric ...
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