检测人中性粒细胞防御素ELISA法的建立及初步应用  

Establishment and application of ELISA for measurement of human neutrophil defensins

作　　者：王永杰[1] 骆晓梅[1] 李晓洲[1] 王涛[1] 齐名[2] 武建国[2] 

机构地区：[1]中国人民解放军第97医院,江苏徐州221004 [2]南京军区南京总医院全军医学检验中心

出　　处：《临床检验杂志》2005年第6期421-422,共2页Chinese Journal of Clinical Laboratory Science

基　　金：江苏省高校自然科学研究计划项目(03KJD310237)

摘　　要：目的建立检测人中性粒细胞防御素(HNP)的ELISA法,并探讨其临床应用价值。方法以纯化的HNP-1为抗原制备特异性抗-HNP单克隆抗体,建立ELISA夹心法,同时检测36名健康献血员血浆中HNP含量,并观察血库4℃保存的全血不同保存时间血浆中HNP含量的变化。结果ELISA法批内和批间变异系数分别为4.8%和9.3%,与进口试剂比较相关性良好(r=0.98,P<0.01),36名健康献血员血浆HNP含量为(46±48)ng/m l。库存全血随保存时间延长血浆HNP含量逐渐升高,保存25天的血浆HNP含量达到760 ng/m l。结论建立的ELISA定量检测法与进口试剂盒有良好的相关性,除血浆外,推测也能适用于细胞培养上清或各种体液中HNP的定量测定。Objective To establish an ELISA method for the measurement of human neutrophil defensins （HNP）, and explore its clinical application. Methods The specific monoclonal antibody against HNP was prepared by using purified HNP-1 as antigen, and an ELISA method was established. The HNP concentrations in plasma from 36 health volunteers were measured, and the changes of HNP content in plasma of stored blood at 4 ℃ was observed. Results The coefficients of variation （CV） of ELISA within the group and between groups were 4.8% and 9.3% respectively. The results of self-made ELISA kit had significant correlation with the results of imported kit （r = 0.98 ,P 〈 0.01 ）. The HNP concentration in the plasma from 36 healthy volunteers was 46 ±48 ng/ml, but the HNP concentration in the plasma from stored blood increased to 760 ng/ml in the blood stored at 4 ℃for 25 days. Conclusions The results of quantitative ELISA method using the prepared kit had significant correlation with the results using imported kit and this method may be used for HNP measurement in the supematant of the cultured cell or various tissue fluids,

关 键 词：酶联免疫吸附试验 防御素 中性粒细胞 

分 类 号：R446.6[医药卫生—诊断学]