Preparation method of direct immunofluorescence detection kit, kit and joint detection method
An immunofluorescence detection and kit technology, applied in the field of immunodetection, can solve the problems of inability to assist diagnosis in time, high experimental environment requirements, and low sensitivity, so as to reduce non-specific reactions of secondary antibodies, shorten operation time, and improve specificity. Effect
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[0061] see figure 1 , which shows a preparation method for a direct immunofluorescence detection kit for joint detection of multiple pathogens according to an embodiment of the present invention, the preparation method comprising the following steps:
[0062] Provide pathogen control slides for multi-pathogen joint inspection;
[0063] Provide specimen sheets for multi-pathogen joint inspection;
[0064] Provide sample diluent;
[0065] Concentrated wash solution for washing pathogen control slides loaded with samples is provided;
[0066] Provides a fluorescein conjugate;
[0067] providing an effective amount of the monoclonal antibody; and
[0068] Provide fluorescent antibody diluent;
[0069] Wherein, the components of the fluorescent antibody diluent include: 0.1% to 0.5% disodium hydrogen phosphate, 0.01% to 0.05% sodium dihydrogen phosphate, 0.5% to 1.5% sodium chloride, 0.01% to 0.05% potassium chloride, 2%-8% disaccharide, 10%-30% glycerin, 0.05%-0.3% casein so...
Embodiment 1
[0131] 1. The method for preparing the pathogen control slide for joint inspection of respiratory infection may further comprise the steps:
[0132] (1) Subculture of cells:
[0133] Method 1 Pathogen-infected cell culture: add nasopharyngeal epithelial cells to be fixed in DMEM medium with 10% fetal bovine serum, and then place them at a temperature of 37°C, a humidity of 90%, and CO 2 Concentration of 5% CO 2 Cultivate in the incubator for 48 hours to recover; then digest and separate with 0.25% trypsin, aspirate the trypsin and count 6 million / T75, and place it again in 5% CO at a temperature of 37°C and a humidity of 90%. 2 Cultured in an incubator for 24 hours for subculture.
[0134] Method for the culture of Legionella pneumophila LP cells: After dissolving the LP special medium powder, carry out autoclave sterilization at 121°C for 15 minutes, pour it into a plate after cooling, make about 15ml of medium in each plate, and wait for the medium to solidify. Inoculate ...
Embodiment 2
[0156] Common reproductive system infections are Chlamydia trachomatis and Ureaplasma urealyticum infection.
[0157] 1. Preparation of pathogen control slides for joint inspection of reproductive system infections is obtained by the following preparation methods:
[0158] (1) Subculture of cells:
[0159] Pathogen-infected cell culture: place the HaCaT cells to be fixed in DMEM medium with 10% fetal bovine serum in 5% CO with a temperature of 37°C and a humidity of 95%. 2 Cultivate in the incubator for 48 hours to recover; after the cells in the culture dish grow into a dense monolayer of cells, digest with 0.25% trypsin, count 6 million / T75, and place them again in a 5 %CO 2 Cultured in an incubator for 24 hours for subculture.
[0160] (2) Pathogen-infected cells:
[0161] The clinical specimens were repeatedly frozen and thawed at -80°C for 3 times to break the cells and release Chlamydia trachomatis, centrifuge at a speed of 3000r / min for 5 minutes, then take the supe...
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