hemolytic Japanese encephalitis complement fixation test

Epidemic encephalitis (J-brain) is caused by Japanese encephalitis virus, which produces hemagglutinin that agglutinates red blood cells such as chicks, pigeons, geese, and sheep. The mature JEV has three structural proteins, V1, V2 and V3. It is proved by the corresponding monoclonal antibody that V3 has four immune-related antigenic spots, one of which is a neutralizing antibody determinant, and the other two are group and Subgroup-specific antigenic determinants, and one with both hemagglutination and neutralizing antigenic determinants. In addition, JEV also has a type-specific soluble complement-binding antigen. JE patients are stimulated by the above-mentioned viral antigens, and hemagglutination inhibitory antibodies, neutralizing antibodies and complement-binding antibodies can be produced in the blood after the disease, and the above three antibodies can be used as a serological diagnostic indicator for the disease, usually by immunization. Detection or immunofluorescence techniques for the detection of specific IgM antibodies have early diagnostic significance. Basic Information Specialist classification: Infectious disease examination and classification: Cerebrospinal fluid examination Applicable gender: whether men and women apply fasting: fasting Tips: If you find suspected symptoms, go to the hospital for an examination. Normal value <1:16. Clinical significance (1) The complement-binding antibody of JE patients generally appears late, and it is reported that the onset of 1-2 weeks, the positive antibody is only 12.8%, only for half a year, the positive antibody of the subject must be newly infected, so the single recovery period Serum greater than or equal to 1:16 is diagnostic. (2) The antibody titer obtained by the general double serum test has a diagnostic value of more than 4 times. If the antibody titer of the double serum is 1:8 or higher, and the serum of the recovery period does not increase by more than 4 times, it is necessary to analyze whether the acute phase specimen takes too late, so it is necessary to re-collect the recovery period blood review. To determine the relationship between antibody titer and this infection. If there is still doubt, it needs to be compared and verified by the neutralization test. High results may be diseases: precautions for epidemic encephalitis B JE neutralizing antibody is an important indicator to confirm JE infection, but its detection is commonly carried out in mice, hamster kidneys and chicken embryo cells. The principle is that the antibody binds to the virus and causes the virus to lose its infectivity. According to the mortality of mice, the lesions of hamster kidney cell culture and the reduction of plaques in chicken embryo monolayers, the antibody titer was measured separately. The method was sensitive, but the operation was troublesome. The general detection was difficult to complete, and it was sent to the professional virus research institute. . Inspection process 1, according to the material: blood 2, the principle of hemolytic encephalitis complement binding test JE patients can produce specific complement-binding antibodies, so the routine complement-binding test is also one of the serological diagnostic indicators for the diagnosis of Japanese encephalitis. 3, reagents (1) The preparation method of the antigen is the same as hemagglutinin, and the normal tissue antigen is prepared by the same method as the healthy rat brain. (2) Mouse anti-JE virus serum generally requires a titer of 1:64 or higher. 4, the operation method (1) Sheep red blood cell suspension, hemolysin and complement should be titrated first, each is formulated to the optimum working concentration. (2) antigen titration, using a dilution of the antigen and antiserum as a square plate titration, the highest dilution of the antigen can be combined with the highest dilution serum and the maximum dilution of the antigen is 1U, such as 1:16 for 1U antigen, The experiment diluted the antigen with 2 U, ie 1:8. (3) Formal test: After the test serum is inactivated at 56 ° C for 30 min, it is first diluted 1:4 or 1:8 to avoid non-specific binding of high concentration serum, followed by dilution, and routine operation according to the complement binding test. Anyone who is not hemolyzed is positive, and hemolysis is negative. The results of the control tube should meet the requirements. The result is calculated as the highest dilution of the test serum that is positive for the complement-binding antibody titer of the test serum. Not suitable for the crowd Those who do not have an indication for examination should not do this check. Adverse reactions and risks Generally no complications and harm.