实验小鼠弓形虫检测方法的建立和应用

Establishment and Application of Methods for the Detection of Antibody to Mouse Toxoplasmosis

目的建立小鼠弓形虫抗体检测方法。方法制备抗原片和可溶性抗原,免疫小鼠制备阳性对照血清,确定了IFA和ELISA检测方法的最适工作条件。根据弓形虫B1基因序列,合成一对寡聚脱氧核糖核苷酸片断,建立PCR检测体系。结果与IHA法相比较,ELISA、IFA的抗体检出率均显著高于IHA,抗体滴度分析,ELISA抗体滴度高于IFA法。对弓形虫感染鼠不同组织进行PCR检测,可检出特异性扩增条带。结论建立的IFA和ELISA检测方法具有灵敏度高、重复性好,结果判定明确等优点,是比较有效的监测方法。建立了弓形虫PCR检测体系,经对弓形虫DNA及病鼠各组织的初步检测,证明本PCR体系具有较高的敏感性和特异性。

Objective To establish the methods for the detection of antibody to mouse Toxoplasmosis. Method Antigen and positive serum of Toxoplasmosis were prepared, and the conditions of IFA and ELISA determined. Two pairs of primers were designed and synthesized based on the sequence of Toxoplasma gondii B1,and the PCR was constructed. Result The positive rates detected by IFA and ELISA were significantly higher than that by indirect haemagglutination （IHA） and antibody titres in ELISA were higher than in IFA. Conclus The IFA and ELISA are the effective methods for detection of mouse Toxoplasmosis with high sensitivity, and PCR can detect Toxoplasma gondii DNA and diagnose toxolpasmosis witb bigb specificity and sensitivity.