A modified ELISA method for cytokeratin 8 detection in the mouse small intestine mucosal injury model:role and value
An improved ELISA method to detect cytokeratin 8(K8)in the mouse intestinal mucosal injury model was evaluated in this study.The mouse intestinal mucosal injury models were established by either enteral injection of sodium formate and H2 O2 or via partial ischemia caused by arteriolar clamps clipping of superior mesenteric artery branches.Pathological changes in intestinal tissue were observed and an optimized ELISA method was used to detect the level of K8 in stools within 24 h after surgery or in serum at different time points,and the change and expression difference of K8 were analyzed.Next,the ROC curve and the AUC were calculated to evaluate the authenticity of the detection method.The sensitivity and specificity were also analyzed to screen for the best cut-off value to provide a reference for clinical detection.The results showed that both enteral injection of sodium formate and H2O2 and arteriolar clamps clipping superior mesenteric artery branches caused significant pathological changes in intestinal tissue.The levels of K8 increased in stools within 24 h after surgery or in serum at different time points after ischemia and reperfusion.The improved ELISA method has high authenticity,and the optimal cut-off values for fecal and serum K8 are 69.78 μg/mL and 0.74 μg/mL respectively,with both high sensitivity and specificity.Utilizing this method to detect the level of K8 in feces or serum of mice reflected the degree of intestinal mucosal damage,therefore providing a reference for clinical monitoring of the occurrence and severity of intestinal mucosal injury,and has high application value.
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