日本血吸虫胞外囊泡蛋白是诊断寄生虫感染的有效生物标记物

Huixin Wu, B. Giri, Huimin Li, Yameng Zheng, Xiaoli Yan, Guofeng Cheng
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摘要

血吸虫是血吸虫病的病原体,分布于世界各地。目前亟需开发一种灵敏的诊断方法来控制这种疾病。此前,我们从日本血吸虫(S. japonicum)中发现了大量胞外囊泡(EV)蛋白,但很少对这些蛋白的诊断潜力进行评估。在本研究中,我们对之前研究中发现的日本血吸虫EV相关蛋白进行了生物信息学分析,然后确定了具有潜在分泌能力的血吸虫特异性蛋白。其中,我们选择了SJCHGC02838蛋白、SJCHGC05593蛋白、SJCHGC05668蛋白和一个假定蛋白(SJHYP),以评估它们检测日本血吸虫感染的诊断潜力。首先,我们利用 qRT-PCR 方法测定了这四种蛋白在转录水平上的表达量,结果表明这些基因在成虫期均有较高的表达量。然后,我们将每种蛋白的全长 cDNA 克隆到原核表达载体中,并成功生成了重组蛋白。纯化重组蛋白后,我们开发了一种间接 ELISA 方法来评估这些纯化重组蛋白的诊断潜力。结果显示,这些蛋白在检测血吸虫感染方面具有很高的灵敏度。此外,这些蛋白还显示出检测血吸虫感染的良好潜力,尤其是 SJCHGC05668 蛋白的早期检测。我们通过 Western 印迹进一步评估了这些重组蛋白的诊断潜力,并用我们之前开发的 cfDNA 方法进行了比较分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Schistosoma japonicum extracellular vesicle proteins serve as effective biomarkers for diagnosing parasite infection
Schistosoma species are the causative agent of schistosomiasis and shows worldwide distribution. There is a great need to develop a sensitive diagnostic approach for controlling the disease. Previously, we identified large numbers of Extracellular Vesicle (EV) proteins from Schistosoma japonicum (S. japonicum), but rarely these proteins have been evaluated for their diagnostic potential. In the present study, we performed bioinformatic analyses of S. japonicum identified EV-associated proteins from the previous study and then identified Schistosoma-specific proteins with potentially secreted capability. Among them, we selected SJCHGC02838 protein, SJCHGC05593 protein, SJCHGC05668 protein and a hypothetical protein (SJHYP) to evaluate their diagnostic potential for detecting S. japonicum infection. First, we determined the expression of these four proteins at the transcript levels using qRT-PCR and revealed that all these genes showed higher expression in adult stage. Then, we cloned the full-length cDNA for each protein into a prokaryotic expression vector and successfully generated the recombinant proteins. Upon the purification of recombinant proteins, we developed an indirect ELISA method to evaluate the diagnostic potential of these purified recombinant proteins. The results showed high sensitivity for detecting Schistosoma infection. Additionally, these proteins also displayed a good potential for detecting Schistosoma infection, especially SJCHGC05668 protein at an early stage. The diagnostic potentials of these recombinant proteins were further evaluated by Western blot and comparatively analyzed by our previously developed cfDNA methods.
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