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
Physiological saline
Centrifuge

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.

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

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).

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