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作 者:呼明闯[1] 汪天平[1] 王恩木[1] 章乐生[1] 司武敏[1] 杨荣笙 罗俊平[1] 郝晓君[1] 翟杜娟
机构地区:[1]安徽省血吸虫病防治研究所,合肥市230061
出 处:《热带病与寄生虫学》2014年第3期140-142,共3页Journal of Tropical Diseases and Parasitology
摘 要:目的建立日本血吸虫抗体检测试剂盒(IHA法)参考品的制备方法。方法采用33.3%的饱和硫酸铵提取日本血吸虫抗体IgGγ球蛋白,经过离子交换剂DEAE层析过滤后,用Lowry法测定蛋白浓度。标定和验证合格后制成冻干阳性标准参考品。采取健康试验兔血清经处理、标定后冻干作为阴性标准参考品。用阴、阳性同批号冻干参考品各100份,分别在1、3、5、7、9、11、13、15、17、19、21、24月后,用ELISA和IHA测定冻干参考品实验室保存条件下的稳定性。结果冻干参考品经过ELISA和IHA两种免疫学方法检测其效价符合要求,其阳性符合率和阴性符合率均为100%,各时间点测定均符合要求。结论本方法制备的阴性和阳性血清冻干品可以作为日本血吸虫抗体检测试剂盒(IHA法)的参考品,其冻干品稳定性满足试剂盒保存两年的要求。Obiective To establish the method of preparing the freeze-dried reference substance of indirect hemagglutination assay kit for the diagnosis of schistosomiasis japonica. Methods Schistosoma japonicum antibodies, IgG γ-globulins were purified by 33.3%saturated ammonium sulfate, then filtered through DEAE exchange chromatography, and finally the protein concentration was tested by Lowry method. It was prepared as positive standard reference substance after calibration and validation. The sera from healthy rabbits after processing and calibration were freeze-dried as negative standard reference substance. The stability in laboratory storage conditions of 100 copies of negative and positive of freeze-dried reference substance were detected respectively after 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, and 24 months by ELISA and IHA. Results The titers of freeze-dried reference substance met the requirements after the test by two kinds of method, and its positive coincidence rate and negative coincidence rate were all 100%. Conclusion The negative and positive freeze-dried sera prepared by this method can be utilized as the reference substance of indirect hemagglutination assay kit, and their stabilities meet the requirements of storing for two years.
关 键 词:日本血吸虫抗体检测试剂盒(IHA法) 阴、阳性冻干参考品 稳定性
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