Complement (Clement) is a multi-molecular system composed of nearly 20 different serum proteins, accounting for about 10% of the total serum globulin. The content of complement in serum is relatively stable and does not increase due to immune response, and it only fluctuates under certain pathological conditions. The basic composition of the complement system includes 9 serum protein components, named C1-9 according to the order of discovery. Complement is a complex reaction system. In addition to lysing cells and assisting sterilization, it can also promote inflammatory reactions. It has a mutually promoting and restrictive relationship with other biologically active systems. The disorder of these relationships is the material basis for many diseases. C3 is a β-globulin. It is the most abundant and important component of complement. It is the central link of the two main activation pathways of complement. It has an important biological activity and can be produced in the liver. C4 is synthesized by the liver and macrophages and participates in the classical activation pathway. It is an important component of complement that is only lower than C3. Commonly used detection methods are rate turbidimetry, rate turbidimetry, etc.
[Detection method] Rate turbidimetry
[Methodological principle] Antigen (C3, C4) and antibody (anti-C3, anti-C4) can react quickly in the liquid phase, and the formed immune complex particles have special optical properties, which make the reaction solution turbid. Rate refers to the rate at which antigens and antibodies form immune complexes per unit time. With the extension of time, the total amount of immune complexes gradually increases, and the rate changes from slow to fast to slow. The time when the reaction rate is the fastest is called the rate peak. When the amount of antibody in the reaction system is excessive, the height of the rate peak is only proportional to the content of C3 and C4. The instrument converts the measured rate peak into the corresponding concentration of C3 and C4 through the corresponding standard curve.
[Sample preparation] Fasting venous blood 2ml, no anticoagulation.
[Reagent] The immunoturbidimeter is equipped with C3 and C4 kits, which contain diluent, buffer, antiserum, etc., and are prepared according to the instructions.
[Instrument] Automatic special protein analyzer.
[Test Step]
1. Specimen processing, centrifugation at 3 500 rpm for 10 min, and separation of serum.
2. Carry out sample determination according to the operating procedure:
(1) Enter the cup number;
(2) Select the tested items C3, C4;
(3) Save;
(4) After programming, place the specimen in the specimen cup and put it in the corresponding position in the specimen tray. Put the reagents of the project in the corresponding position in the reagent tray. After confirming that it is correct, start running the special protein analyzer.
[Normal reference value] C3: 0.8 ～ 1.55g / L5C4: 0.12 ～ 0.36g / L.
[Precautions]
1. When the kit is taken out from the refrigerated environment, it should be balanced at room temperature.
2. When opening the reagent bottle cap, pay attention to breaking the liquid film of the bottle mouth to prevent the liquid sensor from sensing errors.
3. If the serum contains lipids, the lipids should be separated by high-speed centrifugation to prevent false turbidity.
[Clinical Significance] In infectious diseases, C3 and C4 increase during acute infection. Since complement is synthesized in liver cells, severe viral hepatitis and chronic viral hepatitis, especially after conversion to cirrhosis, C3 and C4 levels drop.
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