Detection kit for preeclampsia of pregnant women
A detection kit and a technology for preeclampsia, applied in the field of disease detection, can solve the problems of ratio influence, influence on accurate treatment, influence on the accuracy of the onset risk of eclampsia, etc., achieving simple application, predicting onset risk, and being suitable for large-scale The effect of promoting the application
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Embodiment 1
[0036] Example 1 fluorescent labeling
[0037] Cleaning: Put fluorescent microspheres (Bangs Lab, product number: 11233) into a centrifuge tube, add 0.1M MES (morpholineethanesulfonic acid, pH 5.0) buffer and mix well, and centrifuge at 13000rpm, 15min, 4°C, discard The supernatant was resuspended with 0.1M MES (pH 5.0) buffer for use. Activation and cleaning: the activator EDC (1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride), NHS (1-hydroxypyrroline-2,5-dione) and Fluorescent microspheres are activated according to the mass ratio of 2:1:2, the specific operation is as follows:
[0038] Weigh EDC and NHS and add them to 0.1M MES (pH 5.0) buffer solution to dissolve, quickly take an appropriate amount into the washed fluorescent microspheres, seal them with a parafilm and place them on a 200rpm shaker at room temperature for 30min, take them out at 13000rpm for 30min , centrifuged at 4°C to remove the supernatant, resuspended with an equal amount of 0.1M MES (pH 6...
Embodiment 2
[0045] Spray point of embodiment 2 bonding pad
[0046] The method of spraying the antibody (anti-human placental growth factor) binding pad for PIGF diagnosis is as follows: Fluorescence-labeled antibody solution dilution: the fluorescent-labeled antibody conjugate prepared above was diluted 4 times with fluorescent-labeled antibody resuspension; set point Membrane instrument, turn on the power of the film spotting instrument, set the spraying program, the spraying volume is 8ul / cm; the No. 1 pipe is the spraying channel; the film spotting instrument initialization: put the No. 1 pipe in the fluorescently labeled antibody resuspension solution , select the initialization program, and initialize 6 cycles; spray point: place the bonding pad on the spotting device at a fixed position, press the "GO" key on the control panel to start spraying, remove it after pointing, and check that the spraying point is good Conjugation pads, the fluorescently labeled antibody strips sprayed ev...
Embodiment 3
[0048] The preparation of embodiment 3 nitrocellulose membranes
[0049] Antibody (anti-human placenta growth factor) nitrocellulose membrane preparation method for PIGF diagnosis is as follows: take goat anti-human PIGF polyclonal antibody (Sigma, goat anti-human placenta growth factor polyclonal antibody, 0.1ml / bottle) 500ug, add to In a 5ml graduated centrifuge tube, dilute the antibody to 1ml, and mark the container with a T mark. Take 25ul of goat anti-mouse IgG antibody (Hangzhou Longji Biotechnology, goat anti-mouse IgG, 10mg / bottle), add it to a 5ml graduated centrifuge tube, dilute the antibody to 1ml, and mark the container with a C mark. Set the film spotting instrument, turn on the power supply of the film spotting instrument, set the spray point program, and the spray point volume is 1ul / cm; the No. 1 pipeline is the detection channel with spray points, and the No. 2 pipeline is the control channel with spray point points; the film spotting instrument Initializat...
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