Specimens are most commonly used for plasma or serum, and some specimens are often used for urine, saliva, and feces due to specific test items. At present, the markers for plasma serum samples generally include antigens and antibodies of infectious pathogens, tumor markers, hormones, special proteins, cytokines and therapeutic drugs. Pay attention to the following points in collecting and processing:
First of all, pay attention to avoid serious hemolysis. Hemoglobin contains a heme group, which has a peroxide-like activity. Therefore, in an ELISA assay using HRP as a labeling enzyme, if the hemoglobin concentration in a serum sample is high, it is easily adsorbed during the incubation. In the solid phase, it reacts with the HRP substrate added later to develop color.
Secondly, it is necessary to pay attention to the sample collection and serum separation to avoid bacterial contamination. This is because of the growth of bacteria, some enzymes secreted may decompose the antibody antigen protein; some bacteria endogenous enzymes such as E. coli β - Galactosidase itself produces non-specific interference as determined by the method of labeling with the corresponding enzyme.
Third: Serum samples should be separated by aseptic operation. They can be stored at 2-8 °C for one week. The samples should be stored at -70 °C for a long time. If it is operated by bacteria, it is recommended to freeze it. Fourth: freeze-preserved serum specimens should not be shaken vigorously, mix well, and must avoid repeated freezing and thawing. The mechanical shear force generated by repeated freezing and thawing of the specimen will destroy the molecules such as proteins in the specimen, which may cause false negative results. In addition, freeze-thaw specimens should be mixed.
Fifth: When the specimen is turbid or floc due to bacterial contamination during storage, the supernatant is taken after centrifugation and sedimentation.
First of all, pay attention to avoid serious hemolysis. Hemoglobin contains a heme group, which has a peroxide-like activity. Therefore, in an ELISA assay using HRP as a labeling enzyme, if the hemoglobin concentration in a serum sample is high, it is easily adsorbed during the incubation. In the solid phase, it reacts with the HRP substrate added later to develop color.
Secondly, it is necessary to pay attention to the sample collection and serum separation to avoid bacterial contamination. This is because of the growth of bacteria, some enzymes secreted may decompose the antibody antigen protein; some bacteria endogenous enzymes such as E. coli β - Galactosidase itself produces non-specific interference as determined by the method of labeling with the corresponding enzyme.
Third: Serum samples should be separated by aseptic operation. They can be stored at 2-8 °C for one week. The samples should be stored at -70 °C for a long time. If it is operated by bacteria, it is recommended to freeze it. Fourth: freeze-preserved serum specimens should not be shaken vigorously, mix well, and must avoid repeated freezing and thawing. The mechanical shear force generated by repeated freezing and thawing of the specimen will destroy the molecules such as proteins in the specimen, which may cause false negative results. In addition, freeze-thaw specimens should be mixed.
Fifth: When the specimen is turbid or floc due to bacterial contamination during storage, the supernatant is taken after centrifugation and sedimentation.
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