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目的利用质谱分析技术,对感染弓形虫小鼠血清进行分析,从而发现有诊断意义的差异峰,为弓形虫病提供新的诊断方法。方法实验小鼠共分为2组,弓形虫感染组和空白对照组。分别采集感染弓形虫第10d、14d、35d的小鼠及空白对照组小鼠尾静脉血0.2ml,利用WCX磁珠纯化试剂盒富集血清多肽,应用基质辅助激光解析电离飞行时间质谱(MAIDI-TOF-MS)技术进行俭测,用Autoflexanalysis与Clin ProTools软件对采集图谱进行分析。结果在Mr 1 000～12 000区段,与正常组蛋白指纹图比较,感染组小鼠血清第10d、14d、35d Mr为1 230、1389、1 948的3个多肽峰的表达均显著增加(P<0.05),而Mr为1496的多肽峰的表达显著降低(P<0.05)。结论MALDI-TOF-MS结合磁珠富集技术可发现弓形虫感染小鼠血清的差异多肽,为弓形虫的诊断提供一种新方法。
OBJECTIVE: To analyze the serum of mice infected with Toxoplasma gondii by mass spectrometry, so as to find differential peaks with diagnostic significance and provide a new diagnostic method for toxoplasmosis. Methods Experimental mice were divided into 2 groups, Toxoplasma gondii infection group and blank control group. The mice were infected with Toxoplasma gondii on the 10th, 14th and 35th days, and the tail vein of the blank control group were collected 0.2ml respectively. Serum peptides were enriched by WCX magnetic beads purification kit. The matrix-assisted laser desorption ionization time of flight mass spectrometry (MAIDI- TOF-MS) technology, the analysis of the collected spectra was performed with Autoflexanalysis and Clin ProTools software. Results Compared with the normal histone fingerprints in the range of 1 000 to 12 000 in Mr 1 000, the expression of 3 polypeptide peaks with Mr 1 230, 1389 and 1 948 on the 10th, 14th and 35th days in the infected group were significantly increased P <0.05), while the expression of Mr1496 polypeptide peak was significantly decreased (P <0.05). Conclusion MALDI-TOF-MS combined with magnetic bead enrichment technique can be used to detect the differential polypeptides in the sera of mice infected with Toxoplasma gondii and provide a new method for the diagnosis of Toxoplasma gondii.