cold hemolysis test
cold hemolysis test
Paroxysmal cold hemoglobinuria (PCH) is a kind of autoimmune hemolytic syndrome patients produce a kind of cold-reactive antibody (D-L antibody), this antibody is a kind of IgG, at 37 ℃ and red blood cells can not be firmly bound, when the temperature is as low as 20 ℃ or less, if there is the presence of complement D-L antibody will be able to bind on the surface of the red blood cells, when the temperature is then When the temperature increases to 37℃, due to a series of complement involved in the reaction to form a cavity in the erythrocyte membrane, hemolysis occurs. This experiment is from the Mudanjiang Medical College undergraduate 5-year laboratory guide for laboratory testing majors
Operation method
cold hemolysis test
Principle
Paroxysmal cold hemoglobinuria (PCH) is a kind of autoimmune hemolytic syndrome patients produce a kind of cold-reactive antibody (D-L antibody), this antibody is a kind of IgG, at 37 ℃ and red blood cells can not be firmly bound, when the temperature is as low as 20 ℃ or less, if there is the presence of complement D-L antibody will be able to bind on the surface of the red blood cells, when the temperature is then When the temperature increases to 37℃, due to a series of complement involved in the reaction so that the red blood cell membrane to form a cavity, hemolysis occurs.
Materials and Instruments
guinea pig Move I. Experimental reagents: Complement preparation: extract 2-4 ml of blood from guinea pig's heart. Separate the serum and store it in the refrigerator. Dilute the serum with saline 1:10 for immediate use. Caveat The first and second tubes should be operated below 20℃.Note: Total serum of patients and normal human beings can be inactivated at 56℃ for 30min (the fourth tube). For more product details, please visit Aladdin Scientific website.
Physiological saline
Centrifuge
Experimental operation:
1. Take 8 ml of blood from the user and homozygous or O-type normal human blood, and prepare serum and erythrocyte suspension by the method of removing fibrin with glass beads, or take 6 ml of it without anticoagulation, and divide it into high blood serum. The remaining 2 ml should be anticoagulated to separate the red blood cells.
2. Wash the erythrocytes three times with saline, and finally make 50% erythrocyte suspension.
3. According to the table l-3-7 drops of each liquid, after incubation and centrifugation 1000r/min, 5min observation of hemolysis phenomenon.
Third, the results of the experiment to determine: if the first and second tube hemolysis, the rest of the tube is not hemolysis for positive