斑点金-薄层色谱法检测鼠疫耶尔森氏菌的实验研究  

A study of Dot Immunogold Filtration Assay-Thin Layer Chromatography for the detection of Y.pestis

作　　者：岳明祥[1] 刘楠[1] 宝建喜[2] 张运刚 杨春华 高伟 高志贤[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所 [2]中国疾病预防控制中心鼠疫布氏菌病预防控制基地 [3]解放军63850部队防检所

出　　处：《中国地方病防治》2005年第6期321-323,共3页Chinese Journal of Control of Endemic Diseases

基　　金：国家自然科学基金项目(No.30371218)

摘　　要：目的建立一种简便、快速的斑点金-薄层色谱法(DIGFA-TLC)用于鼠疫耶尔森氏菌的快速定量诊断。方法用柠檬酸钠还原法制备10 nm的胶体金标记鼠疫耶尔森氏菌多价抗体,制备成鼠疫抗体金探针,通过薄层色谱扫描对样品斑点进行定量测定。结果斑点金-薄层色谱法对鼠疫FI抗原的最低检出限为5 ng;对EV76的最低检出限为1.2×105cfu;且二者呈高度的正相关关系,前者相关直线方程为y=448.467 37+2.511 43x,后者相关直线方程为y=375.760 88+4.112 31×10-4x;整个反应过程在10 min内完成。结论该方法简便、快速、特异、敏感,可以实现鼠疫耶尔森氏菌的定量监测。Objective A rapid,simple detective method - dot immunogold filtration assay - thin layer cbromatography（DIGFA - TLC）tor quantitative detecling of Plague was established in this experimental study. Methods 10nm colloidal gold marked polyelonai anti - PlagueFl was prepared by reduction process with sodium citrate to fabricate a probe of anti - PlagueFI colloidal gold for quaulila- tive measurement of the immuno dot of sample witb TLC. Results The detective limit of FI - anligeu by DIGFA - TLC was as low as 5ngand that stains of EV76 was 1.2 × 10^5 efu; furthermore, high positive correlation was between them. The correlative liuear equations were y=448.467 37＋2.511 43x, P 〈0.01 and y=375.760 88＋4.112 31 × 10^-4x, P 〈0.05 respectively. The whole reaetive process could be finished within 10 rain. Conclusions The DIGFA - TLC is a simple, rapid, specific, sensitive and reliable assay, and quantitative detection of the Plague can be realized.

关 键 词：斑点金 薄层色谱 鼠疫耶尔森氏菌 

分 类 号：R516.8[医药卫生—内科学]