Establishment Of Method For Detecting Pepsin Content And Its Preliminary Clinical Application

ObjectiveTo establish double antibody sandwich biotin-streptavidin chemilumine-scence enzyme immunoassay method for detection of pepsin content; detection of the clinical specimens by this method and to investigate its clinical value.MethodsBased on double antibody sandwich principle,using rabbit anti-human pepsin antibody coated Solid-phase microplates,combined with biotin-streptavidin system,pepsin antibody labeled with biotin.steptavidin coupled HRP,application CLEIA.to eastablish the method of determination of pepsin content.The experimental conditions of the double antibody sandwich biotin-streptavidin chemiluminescence enzyme immunoassay method was optimized and the experimental sensitivity,specificity,accuracy was evaluated. Various gastric juice samples were collected including normal controls (21cases),gastritis (44cases),reflux esophagitis (15cases),gastric ulcer (13cases),duodenal ulcer (15cases),gastric carcinoma (5cases) the method was employed to detect the pepsin content in various gastric juice.Explore the impact of different types of the disease, the pH of gastric juice,patient gender and age on pepsin content.ResultsCoating concentration of pepsin antibody was5μg/ml,the dilute concentrations of biotin-labeled antibody was1:500, at37℃the reaction time was60min,30min,30min respectively,the chemiluminescent substrate reaction time was5min.The linear rang was2-200ng/ml.Correlation coefficient was0.97. The sensitivity was2.75ng/ml.The crossover rate was<0.5%with lysozyme.salivary amylase.pepsinogen I and pepsinogen II.The coefficient of variation of inter-and intra-assay was<7%and<10%. Pepsin content of gasric juice was detected. The samples were from113cases of patient with different diseases.The data was analyzed by spss17.0.The results were as follows: Comparing with the normal controls, the pepsin content of gastric ulcer and duodenal ulcer were higher, the pepsin content of gastric carcinoma were lower; there were no significant differences in pepsin content of the normal controls with gastritis and reflux esophagitis The lower of the pH, the higher of the pepsin content;there was no significant difference between male and female no significant difference among the youth.middle age, older age group.ConclusionsThe advantage of double antibody sandwich biotin-streptavidin chemiluminescence enzyme immunoassay method was easy to operate, high sensitivity and specificity. This detection method could be widely used in clinical research.The pepsin content of the gastric juice related to the type of different gastrointestinal disease and pH,has nothing to do with sex and age.