日本血吸虫感染兔血清蛋白质质谱分析

Mass spectrometry analysis of serum proteins from Schistosoma japonicum infection rabbits

目的通过对感染日本血吸虫兔血清进行蛋白质质谱分析，确定有潜在诊断意义的标志蛋白，为日本血吸虫病血清学诊断提供理论依据。方法取不同感染时期的兔血清，利用WCX磁珠纯化试剂盒富集血清蛋白，应用基质辅助激光解吸电离飞行时同质谱（MAIDI—TOF-MS）技术进行分析。结果与正常兔血清蛋白指纹图谱比较，在Mr1000-12000区段，不同感染时期实验兔血清分析共有63个蛋白峰，其中7种蛋白有显著差异，利用BrukerClinProTools软件分析，在感染后第6天，Mr1787蛋白有显著上调（P〈O．05）；感染后第9天，Mr2834蛋白有极显著上调（P〈0．01），另外Mr3483蛋白有显著上调（P〈0.05），Mr4018蛋白有显著下调（P〈0.05）；在感染后第12天，Mr3151蛋白有显著下调（P〈0.05），而Mr4018蛋白在感染后第12天，则出现极显著下调（P〈0.01），两者趋势一直持续至感染后第30天。家兔感染后第9～12天，血清中Mr3530蛋白有显著上调（P〈0.05），Mr1716蛋白显著下调（P〈0.05）；第12~30天血清中Mr3151、3530蛋白有显著下调（P均〈0.05）。结论MALDI-TOF-MS结合磁珠富集技术可发现及检测日本血吸虫病具有潜能的早期生物标志物，为从蛋白质水平研究日本血吸虫病免疫诊断及寻找新的治疗靶位奠定了基础。

Objective To search for a biomarker from the serum of Schistosoma japonicum infected rabbits for early diagnosis of schistosomiasis. Methods The sera were obtained from different periods of the infected rabbits. The serum proteins were generated by WCX kit （Bruker Daltonics GmBH） and analyzed by the technique of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Results In the mass range from 1 000 to 12 000 Da, sixty-three proteins were captured by WCX kit. ClinProTools software was used to find the differential expressed proteins. The result revealed 7 distinct proteins compared with normal serum. Among them, 1 787 Da protein expression was increased （P〈0.05） after 6 days from Schistosorna japonicum infection. After 9 days, 2 834 Da protein was increased significantly（P〈0. 01）, 3 483 Da protein was increased and 4 018 Da protein decreased （P〈0. 05）. After 12 days, 3 151 Da protein expression was decreased （P〈0. 05）, whereas 4 018 Da protein decreased significantly （P〈0.01）. From 9 to 12 days, 3 530 Da protein was increased and 1 716 Da protein decreased （P〈0.05） ; from 12 to 30 days, 3 151 Da and 3 530 Da proteins were decreased （P〈0.05）. Conclusions The technology of MALDI-TOF-MS combined with magnetic beads can be a desired method in detecting the biological markers in the earli- er period of schistosomiasis. The biological tag-protein has a potential value in the early diagnosis of schistosomiasis.