日本血吸虫NP30抗体检测方法的优化  

Optimization of Schistosoma japonicum NP30 antibody detection method

作　　者：许宁[1] 吴玉龙[2] 仇镇宁[1] 陈凤 李远林 方文 杨杰 冯振卿[1] 管晓虹[1] 

机构地区：[1]南京医科大学卫生部抗体技术重点实验室,南京210029 [2]滨州医学院病原生物学教研室 [3]云南省大理白族自治州血吸虫病防治研究所 [4]云南省大理白族自治州巍山县血吸虫病防治研究所

出　　处：《中国血吸虫病防治杂志》2006年第2期88-92,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家高技术研究发展计划(863计划)(2002AA214181)

摘　　要：目的为了提高单克隆抗独特型抗体NP30抗体检测系统在大山区日本血吸虫病疫区的诊断效能,在原有的双抗体夹心ELISA方法的基础上对其进行优化、改良,建立间接ELISA血清学诊断方法。方法通过正交设计,选用L27(313)正交表,对NP30腹水包被酶标板的浓度、血清浓度、血清孵育时间和辣根过氧化物酶标记抗人IgG的二抗孵育时间共4个因素的3个水平进行不同组合的实验,确定该间接法的最佳实验条件。并用建立的方法对50份云南大山区日本血吸虫粪孵阳性病人血清样本和50份非疫区正常人血清样本进行检测,评价其敏感性和特异性。结果确定的该间接法最佳检测条件为:NP30腹水4μg/ml包被酶标板,血清浓度为1∶100,37℃孵育60min,二抗为37℃孵育30min。该方法的敏感性和特异性分别为88%和96%。结论该方法具有较高的诊断价值,可用于大山区日本血吸虫病疫区的诊断检测。Objective To set up an indirect ELISA serum diagnostic method based on the original NP30 antigen-sandwiched ELISA, which can enhance its diagnostic efficiency when applied in schistosomiasis epidemic spots of great mountainous areas in Yunnan Province. Methods According to the rules of orthogonal test, table L27（31^13） was chosen to determine the appropriate values of four factors, which were the concentration of coating antigen （NP30 ascites）, the concentration of serum, the cultivating time of serum and the cultivating time of HRP-anti-human IgG. The improved method was used to detect 50 serum samples from the patients with schistosomiasis and 50 ones from the healthy people to estimate its sensitivity and specificity. Results The best values of the four factors were 4 μg/ml, 1 ： 100, 60 min and 30 min, respectively. The sensitivity of the method was 88% as well as its specificity 96%. Conclusion This optimized ELISA method is suitable for application in great mountain epidemic areas with higher diagnostic efficiency.

关 键 词：日本血吸虫病 正交试验 抗独特型抗体NP30 酶联免疫吸附试验 

分 类 号：R532.21[医药卫生—内科学]