PDF(Pubmed)
Abstract：
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.
摘要：
血吸虫是血吸虫病的病原体，并在世界范围内分布。非常需要开发用于控制疾病的灵敏诊断方法。以前,我们从日本血吸虫中鉴定出大量的细胞外囊泡（EV）蛋白（S.japonicum),但很少对这些蛋白质的诊断潜力进行评估。在本研究中,我们对日本血吸虫进行了生物信息学分析,从之前的研究中鉴定出EV相关蛋白,然后鉴定出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蛋白和一种假想蛋白（SJHYP），用于评估其检测日本血吸虫感染的诊断潜力。首先,我们使用qRT-PCR确定了这四种蛋白质在转录水平上的表达，并揭示了所有这些基因在成年阶段都显示出更高的表达。然后，我们将每种蛋白的全长cDNA克隆到原核表达载体中，并成功地产生了重组蛋白。重组蛋白纯化后，我们开发了一种间接ELISA方法来评估这些纯化的重组蛋白的诊断潜力。结果对检测血吸虫感染具有较高的敏感性。此外,这些蛋白质也显示出检测血吸虫感染的良好潜力，特别是早期的SJCHGC05668蛋白。通过Western印迹进一步评估这些重组蛋白的诊断潜力，并通过我们先前开发的cfDNA方法进行比较分析。
