Thin Layer Immunoassay: An Economical Approach to Diagnose Helicobacter pylori Infection in Gastroduodenal Ulcer Disease Patients of Pakistan, a Comparative Analysis

薄层免疫测定:诊断巴基斯坦胃十二指肠溃疡病患者幽门螺杆菌感染的经济方法,比较分析

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作者:Faisal Aziz, Shahana Urooj Kazmi

Abstract

Helicobacter pylori is a causative agent of gastritis, gastroduodenal ulcers and gastric adenocarcinoma. The majority of H. pylori-associated patients live in underdeveloped areas, facing the problem of lack of proper diagnostic facility. Hence, a simple and economical assay is required to handle the majority of gastric patients. Serum samples from gastroduodenal ulcers and gastritis patients were screened for H. pylori infection by thin layer immunoassay. A polystyrene plate coated with H. pylori sonicate whole cell antigen (10 µg/mL). Two-fold diluted patient's serum was allowed to react at 37 °C, incubated at 60 °C for 1 min over a water bath and the water condensation pattern for the H. pylori antibody was recorded. ELISAs were used as reference assays to evaluate the efficacy of the developed thin layer immunoassay (TLI). Gastric patients' blood samples (62% male and 6% female) tested positive for H. pylori, while age-wise, 15-25-year-old males (36%) and 65-75-year-old females (50%) showed the highest number of H. pylori infections. TLI showed sensitivity (72-67%), specificity (100%), accuracy (94-69%) and κ value (0.493-0.357) in comparison with wELISA (Surface whole cell ELISA), sELISA (sonicate whole cell ELISA) and kELISA (commercial KIT ELISA). We conclude that thin layer immunoassay is a low cost, fast, simple and clinically reliable method for H. pylori diagnosis at initial stages in patients in under-developed countries.

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