基于IgG4介导的蛋白质组学日本血吸虫诊断标志物的筛选及应用-病原生物学专业论文.docxVIP

ten different proteins. These proteins were S. japonicum aconitase (SjAA), heat shock protein 70 (HSP70), pyruvate kinase M2 isoform (SjPKM2), phosphoglycerate kinase (SjPGK1), α-L-fucosidase (SjALF), inorganic pyrophosphatase (SjIPP), glutathione transferase (SjGST), 21.7 kDa antigen (Sj21.7), tegument membrane-associated antigen (Sj22.6) and sarcoplasmic calcium-binding protein of S. mansoni (SmSCP). Bioinformatics analysis displayed that SjIPP possesed variety of biological functions and abundant antigen epitopes for B cell, which implied potential insight about diagnosis for schistosomiasis. Furthermore, SjIPP was successfully expressed, and the recombinant protein was further applied in the diagnosis of human schistosomiasis using IgG4-ELISA. The ELISA results revealed sensitivities of 100% and 87.1% for acute schistosomiasis and chronic schistosomiasis, whereas the assays showed a specificity of 95.0% with rSjIPP recombinant protein. Remarkably, rSjIPP showed low cross reaction with these sera infected with Paragonimus westernami(lung fluke), C. sinensis (liver fuluke)or nematodes(roundworms). The cross reaction rates were respectively 3.7%, 4.5% and 4.2%. Conslusions SjIPP was successfully screened as a diagnostic candidate from AWA of Schistosoma japonicum by IgG4-oriented proteomics followed by expression. Our present study confirmed that rSjIPP was a useful diagnostic biomarker for Schistosoma japonicum infection. Keywords: Schistosoma japonicum; proteomics; IgG4; inorganic pyrophosphatase (IPP); diagnosis 前 言 血吸虫病依然是世界上危害公众健康最严重的疾病之一[1]。引起血吸虫病的 主要病原体有曼氏血吸虫、日本血吸虫、埃及血吸虫等。目前全球血吸虫病患者 超过 2 亿，风险人群超过 7 亿[2, 3]（表 1，图 1），每年因血吸虫病死亡人数 28 万之多。我国是日本血吸虫病流行区，解放后，由于我国各级政府的高度重视， 血吸虫病的流行得到了很好的控制，甚至有几个省或直辖市血吸虫病已经消除， 我国血吸虫病疫情目前已基本达到控制阶段[4]，提高监测敏感性为导向的低流行 区监测技术和方法，以达到有效控制血吸虫病疫情，防止疫情回升和反弹。 表 1：全球血吸虫感染及风险人群分布（2008 年 WHO/NTD 报道） 在过去半个多世纪，我国血吸虫病的防治取得了重大的进步，目前多数地区 流行状态处于相对较低的水平，但在一些核心地区的局部区域，血吸虫病流行依 然存在，这些地区再次感染和反复化疗还时有发生。选择性地服用吡喹酮是目前 我国血吸虫病控制项目的重要策略之一，而血吸虫病的正确诊断是血吸虫病控制 项目实施的第一步，