CLIA Reagents of Inflammation (Lp-pla2)

Lp-PLA2 is one of the isoforms in the phospholipase superfamily, also known as platelet-activating factor acetylhydrolase, secreted by macrophages, T cells and mast cells in the vascular intima. Lp-PLA2 expression was upregulated in atherosclerotic plaques and was strongly expressed in macrophages in vulnerable plaque fibrous caps.

Lp-PLA2 can hydrolyze oxidized phospholipids in oxidized low-density lipoprotein ox-LDL to generate lipid pro-inflammatory substances such as lysolecithin and oxidized free fatty acids, which in turn produce a variety of atherogenic effects, including endothelial cell death and Endothelial dysfunction stimulates the production of adhesion factors and cytokines. These substances can further generate a self-reinforcing cycle by chemotactic inflammatory cells, producing more pro-inflammatory substances.

Lp-PLA2 released into the blood circulation is mainly combined with apolipoprotein (Apo) B-rich lipoprotein, low density lipoprotein (LDL) accounts for 80%, and the rest is combined with high density lipoprotein (HDL), lipoprotein and polar lipoprotein. Low density lipoprotein (VLDL) binding. In patients with atherosclerotic disease, Lp-PLA2 levels were positively correlated with LDL subfraction levels.

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The detection of Lp-PLA2 has two kinds of activity method and concentration method. Activity methods mainly use high performance liquid chromatography, radioactivity assay and enzymatic hydrolysis of substrates. High-performance liquid chromatography has low sensitivity and is susceptible to interference from various components in blood.

The radioactivity determination method has problems such as radioactive contamination of reagents, low accuracy, and poor repeatability; the enzymatic hydrolysis of substrates mainly relies on imported reagents, which has the problem of high cost. In clinical practice, the main method used to detect the concentration is ELISA. The EUSA method adopts the solid-phase sandwich method, and the standard substance with known Lp-PLA2 concentration and the sample with unknown concentration are added to the microtiter plate for detection.

Lp-PLA2 and biotin-labeled antibodies were first incubated simultaneously. After washing, avidin-labeled HRP was added. After incubation and washing, unbound enzyme conjugates are removed, and then substrates A and B are added, and the enzyme conjugates act simultaneously. produce color. There is a positive correlation between the depth of color and the concentration of Lp-PLA in the sample.