In recent years, enzyme immunoassay technology has developed rapidly. The methods of HCG detection are endless. At present, the detection methods applying enzyme immunoassay technology include: spotting, double-antibody sandwich method, ABC-ELISA method, etc. The commonly used ones are introduced as follows, and this experiment is from Mudanjiang Medical College, undergraduate 5-year clinical laboratory instruction.
Operation method
Enzyme-linked immunoassay for human chorionic gonadotropin assay
Principle
Enzyme spotting assay (membrane EIA) experimental principle: the enzyme spotting method for measuring β-HCG in urine is an enzyme immunoassay combining monoclonal and polyclonal antibodies, and the principle is as follows: the reaction system is a membrane treated with anti β-HCG antibody, and when the urine passes through the upper layer of the filter membrane to reach the reaction membrane, there are HCG molecules on the membrane, and then they bind specifically to another exposed site on the surface of the membrane with the anti-β-HCG antibody. It will be combined with the anti-β-HCG antibody on the membrane, so when adding the enzyme-labeled HCG antibody complex, it will be specifically combined with another exposed site on the membrane surface of HCG molecules, at this time, the membrane will be peeled off, add the enzyme substrate coloring solution, then the reaction system is a membrane treated with anti-β-HCG antibody. At this point, the membrane is peeled off and the enzyme substrate is added to color the complex, at which time different colored dots will appear.
Materials and Instruments
Substrate chromogenic solution HCG standard solution Enzyme-beta-HCG antibody complex Move I. Experimental reagents: For more product details, please visit Aladdin Scientific website.
1, enzyme-labeled-β-HCG antibody complexes
2, substrate color development solution
3、HCG standard solution
4、Negative control
5、Reaction system:
Coated anti-HCG antibody, the above reagents should be stored in the refrigerator at 2-8 ℃, do not mix the reagents of different batches. When the kit is taken out of the refrigerator for use, it should be placed in room temperature (18-3O ℃) for 20min.
Experimental specimen collection: any random urine can be used to measure the method, but the first morning urine is better, collect urine specimens can be any clean, dry plastic or glass containers if the specimen can not be immediately on the specimen immediately on the specimen, can be placed in the refrigerator at 2-8 ℃ for 48 hours specimens in the test before the specimen must be placed at room temperature (18-30 ℃) and parked for 20 minutes, do not have to be, and do not have to be filtered.
Second, the experimental method:
The urine sample or HCG standard is filled with a small pool at the upper end of the reactor, to be completely absorbed and then add 3 sober labeled HCG antibody complex in the small pool on the membrane, to be completely absorbed for 1 minute after the tip of the liquid tube with the stripping membrane, and finally add 3 substrate color solution in the reaction membrane, observe the results on the membrane, the specific operation of the specific instructions.