摘要
目的建立基于麻疹病毒(Measles virus,MV)IgG抗体酶联免疫吸附试验(ELISA)试剂盒的尿素变性亲和力检测方法。方法选取2018-2019年广西MV IgG阳性的麻疹病例血清标本,采用亲和力试剂盒检测MV IgG亲和力。选择低和高亲和力血清标本各6份,在ELISA酶标板中加入尿素处理后,检测IgG浓度,计算IgG浓度下降率;在亲和力试验中改变尿素条件,计算高低亲和力标本的相对亲和力指数(Relative avidity index,RAI)比值,确定本研究方法的最优实验条件。选择100份血清标本同时采用本研究建立的方法和成品试剂盒检测MV IgG亲和力,计算检测一致率。结果6份低亲和力和6份高亲和力血清标本RAI分别为24%-39%、67%-86%;当4mol/L(pH6.4-7.4)、5mol/L(pH6.4-7.4)和6mol/L(pH6.4-7.0)的尿素处理酶标板后,MV IgG浓度下降率≤48.67%;当尿素条件为4mol/L(1-40min)、5mol/L(1-20min)、6mol/L(1-5min)时,IgG浓度下降率≤47.53%;尿素条件为6mol/L(pH7.0)和6mol/L(5min)时RAI比值分别为2.30和2.43。本研究亲和力试验的尿素处理最优条件为6mol/L(pH7.0)的尿素在37℃作用5min;该方法与成品试剂盒的亲和力检测一致率为92%。结论本研究建立的基于MV IgG抗体ELISA试剂盒的尿素变性亲和力检测方法简便可行且结果可靠。
Objective To establish a method for assaying urea denaturation avidity based on the enzyme-linked immunosorbent assay(ELISA)for measles virus(MV)IgG antibody.Methods We selected serum samples of MV IgG positive measles cases obtained during 2018-2019 in Guangxi to test for avidity using an avidity test kit.We tested for IgG concentration in 6 low-and high-avidity serum samples treated with urea in the ELISA plates to determine decreases of IgG.We varied urea conditions in the avidity test to calculate a relative avidity index(RAI)ratio of high versus low avidity sera to identify optimal experimental conditions of the technique.We tested 100 serum samples for MV IgG avidity using both the method we are establishing and the avidity test kit to calculate the concordance of the two method.Results RAIs were 24%-39%for the 6 low-avidity serum samples and 67%-86%for the 6 high-avidity serum samples.When urea conditions were 4 mol/L(pH6.4-7.4),5 mol/L(pH6.4-7.4),or 6 mol/L(pH6.4-7.0),the IgG concentration decreases were all≤48.67%.When urea conditions were 4 mol/L(1-40 minutes),5 mol/L(1-20 minutes),and 6 mol/L(1-5 minutes),the IgG concentration decreases were≤47.53%.The RAI ratio of the avidity test was 2.30 for 6 mol/L(pH7.0)and 2.43 for 6 mol/L(5 minutes).The optimal urea condition was when serum samples were treated with urea,6 mol/L(pH7.0),at 37℃for 5 minutes.The avidity concordance rate was 92%between the method we are establishing and the avidity test kit.Conclusions The detection method of urea denaturation avidity based on MV IgG antibody ELISA was simple,feasible,and reliable.
作者
秦月
韦一知
邓丽丽
韦敬航
梁亮
马宇燕
刘巍
Qin Yue;Wei Yizhi;Deng Lili;Wei Jinghang;Liang Liang;Ma Yuyan;Liu Wei(Guangxi Zhuang Autonomous Region Center for Disease Control and Prevention,Nanning 530028,Guangxi,China)
出处
《中国疫苗和免疫》
CSCD
北大核心
2022年第1期30-34,40,共6页
Chinese Journal of Vaccines and Immunization
基金
2016年广西卫生健康委员会自筹科研课题(Z2016444)。
