Leptospira complement fixation assay

Leptospirosis is a natural epidemic disease caused by various pathogenic spirochetes. There are many groups and serotypes of pathogenic Leptospira, and the group-specific chemical composition is a lipopolysaccharide complex. The chemical composition of the type-specific is a proteoglycan complex. The polysaccharide composition of different types of Leptospira differs. The patient can produce the corresponding antibody, and examining the specific antibody can help the diagnosis of the disease. Basic Information Specialist classification: Infectious disease inspection and classification: pathogenic microorganism inspection Applicable gender: whether men and women apply fasting: fasting Analysis results: Below normal: Normal value: no Above normal: negative: The normal value is negative. Positive: A serum dilution of 1:100 or greater than 1:100 indicates that the indirect fluorescent antibody staining positive can be judged as positive, suggesting a leptospirosis infection. Tips: Do not eat too greasy, high-protein foods the day before the blood draw, avoid heavy drinking. The alcohol content in the blood directly affects the test results. Normal value negative. Clinical significance Abnormal results: A serum dilution of 1:100 or greater than 1:100 indicates that the indirect fluorescent antibody staining is positive and can be judged as positive. For example, if the titer of the second blood collection is more than 4 times, the clinical significance is greater. People who need to check: There are high fever, fatigue, weakness, body aches, conjunctival hyperemia, diaphragmatic tenderness and other symptoms; accompanied by diffuse lung bleeding, obvious liver, kidney, central nervous system damage and other symptoms. Positive results may be diseases: leptospirosis, leptospirosis, leptospirosis, renal damage, leptospirosis, nervous system performance considerations Before inspection: 1, do not eat too greasy, high-protein food the day before the blood, to avoid heavy drinking. The alcohol content in the blood directly affects the test results. 2. After 8 pm on the day before the medical examination, you should start fasting for 12 hours to avoid affecting the test results. 3, should relax when taking blood, to avoid the contraction of blood vessels caused by fear, increase the difficulty of blood collection. After inspection: 1. After blood is drawn, local compression is required at the pinhole for 3-5 minutes to stop bleeding. Note: Do not rub, so as not to cause subcutaneous hematoma. 2, the pressing time should be sufficient. There is a difference in clotting time for each person, and some people need a little longer to clotting. Therefore, when the surface of the skin appears to be bleeding, the compression is stopped immediately, and the blood may be infiltrated into the skin due to incomplete hemostasis. Therefore, the compression time is longer to completely stop bleeding. If there is a tendency to bleed, the compression time should be extended. 3, after the blood draw symptoms of fainting such as: dizziness, vertigo, fatigue, etc. should immediately lie down, drink a small amount of syrup, and then undergo a physical examination after the symptoms are relieved. 4. If there is localized congestion, use a warm towel after 24 hours to promote absorption. Inspection process 1, the principle of the leptospires complement binding test After the Leptospira antigen binds to the corresponding specific antibody, a fluorescein-labeled anti-human globulin antibody is added to fluoresce under a fluorescence microscope. According to whether or not fluorescence is present, it is known whether or not there is a specific antibody against Leptospira in the serum to be tested. 2, reagents (1) Fluorescein-labeled anti-human IgG. (2) Leptospira antigen. The Leptospira cultures of 4 to 5 serotypes can be mixed in equal amounts. (3) Preparation of Leptospira antigenic tablets. The Leptospira antigen was taken in small amounts on the inoculating loop and applied to a thin and clean glass slide. Each slide was coated with 2 rows of 16 points, naturally dried, and fixed with acetone for 5 min at room temperature for use. 3, the operation method (1) The test serum inactivated at 56 ° C for 30 min was diluted with phosphate buffered saline (pH 7.2) to a ratio of 1:50 to 1:400 for a total of 4 dilutions. (2) Each test serum of each dilution is added dropwise to each point of the antigen smear, and 4 test serums can be dropped for each smear. The inside of the humidification box was applied at 37 ° C for 30 min. (3) Pour off the remaining serum on the slide. Wash with phosphate buffered saline (pH 7.2). (4) Add a few drops of fluorescein-labeled anti-human IgG antibody and let it stand at 37 ° C for 30 min. (5) Wash with the same method, wait until dry. The buffer was fixed with buffered glycerol (pH 9.5) and examined by fluorescence microscopy. (6) Judgment of results: (3+) The cells expanded slightly and showed a bright pale green fluorescence. (2+) The cells are clear and have a bright pale green fluorescence. (+) The cells can be clearly distinguished, but the fluorescence is slightly poor. (±) The cells are blurred or atypical, and the brightness is poor. (-) No cells were visible. (+) is the end point, and (±) and (-) are negative. 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.