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将蛋白酶K处理的细胞用作脂多糖抗原用于幽门螺杆菌感染的血清学诊断

Utilization of proteinase K-treated cells as lipopolysaccharide antigens for the serodiagnosis of Helicobacter pylori infections.

作者信息

Amano K, Yokota S, Ishioka T, Hayashi S, Kubota T, Fujii N

机构信息

Central Research Laboratory, Akita University School of Medicine, Akita, Japan.

出版信息

Microbiol Immunol. 1998;42(7):509-14. doi: 10.1111/j.1348-0421.1998.tb02317.x.

Abstract

We have evaluated the use of proteinase K (PK)-treated cells isolated from Helicobacter pylori as lipopolysaccharide (LPS) antigens in an immunoblot assay and an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of H. pylori infection. The sera from patients with chronic gastritis, gastric ulcer, duodenal ulcer or gastric cancer, and from healthy adults with or without H. pylori infection were assayed with three commercial serodiagnostic kits (HM-CAP, Helico-G, and G.A.P. II) and novel methods relying on the use of PK-treated cells. The PK-treated cells used in these assays were selected on the basis of their possibility to possess a common epitope in the O-polysaccharides of H. pylori, which is known to be highly immunogenic in humans. Of the sera from these patients, 71-94% were positive with the commercial kits, 97% with immunoblot assay, and 90% with ELISA. On the other hand, of the healthy adults infected with H. pylori, 72-97% were positive with the commercial kits, 86% with immunoblot assay, and 72% with ELISA. PK-treated cells that did not contain the common epitope were unsuitable as an antigen for immunoblot assay or ELISA. Furthermore, the reactivity of these sera reacted specifically with H. pylori PK-treated cells but not with LPSs from other gram-negative bacteria, such as Campylobacter, Proteus, Bordetella, and Salmonella. These results demonstrate that the serological assays relying on the use of H. pylori PK-treated cells possessing a highly antigenic epitope are potentially useful as a serodiagnostic test for H. pylori infection.

摘要

我们评估了将经蛋白酶K(PK)处理的幽门螺杆菌分离细胞作为脂多糖(LPS)抗原,用于免疫印迹分析和酶联免疫吸附测定(ELISA)以进行幽门螺杆菌感染血清学诊断的情况。使用三种商业血清学诊断试剂盒(HM-CAP、Helico-G和G.A.P. II)以及依赖于使用经PK处理细胞的新方法,对患有慢性胃炎、胃溃疡、十二指肠溃疡或胃癌的患者以及有无幽门螺杆菌感染的健康成年人的血清进行了检测。这些检测中使用的经PK处理细胞是根据它们在幽门螺杆菌O-多糖中可能拥有共同表位而选择的,已知该表位在人类中具有高度免疫原性。在这些患者的血清中,71%-94%使用商业试剂盒呈阳性,97%使用免疫印迹分析呈阳性,90%使用ELISA呈阳性。另一方面,在感染幽门螺杆菌的健康成年人中,72%-97%使用商业试剂盒呈阳性,86%使用免疫印迹分析呈阳性,72%使用ELISA呈阳性。不包含共同表位的经PK处理细胞不适用于免疫印迹分析或ELISA作为抗原。此外,这些血清的反应性与幽门螺杆菌经PK处理细胞特异性反应,但与其他革兰氏阴性菌如弯曲杆菌、变形杆菌、博德特氏菌和沙门氏菌的LPS无反应。这些结果表明,依赖于使用具有高度抗原性表位的幽门螺杆菌经PK处理细胞的血清学检测作为幽门螺杆菌感染的血清学诊断试验可能是有用的。

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