应用胶体金免疫层析技术快速检测杜克雷嗜血杆菌方法的建立  

Development of A Rapid Gold-Immunochromatography Assay for Haemophilus Ducreyi

作　　者：王虹[1,2] 张辛艳[1] 段琼[1] 刘兵[1] 冯丹红[1] 

机构地区：[1]吉林省人民医院皮肤性病科,吉林长春130021 [2]吉林大学第一医院皮肤性病科,吉林长春130021

出　　处：《中国皮肤性病学杂志》2009年第7期446-448,共3页The Chinese Journal of Dermatovenereology

基　　金：吉林省科技厅;吉林省科技发展计划资助项目(20010421)

摘　　要：目的建立快速检测杜克雷嗜血杆菌的胶体金免疫层析试验(GICA)系统。方法采用胶体金颗粒标记抗rHgbA-IgG抗体,制备出免疫胶体金;采用双抗体夹心模式建立适用于杜克雷嗜血杆菌检测的胶体金免疫层析技术;以ELISA试验作为平行对照,对其检测的灵敏度及特异性进行评价。结果以该方法检测杜克雷嗜血杆菌、流感嗜血杆菌以及其他7种生殖器溃疡相关细菌,杜克雷嗜血杆菌显示强阳性,其余8种均为阴性,无交叉反应发生,GICA与ELISA实验结果相符合;GICA和ELISA检测灵敏度分别为:纯化抗原实验GICA为6.25ng/mL,ELISA为1.56ng/mL;细菌学实验GICA为2×106cfu/mL,ELISA为2×105cfu/mL;模拟样本检测实验GICA为1×107cfu/mL,ELISA为1×106cfu/mL。GICA能在15min内完成检测,稳定性良好。结论建立的检测杜克雷嗜血杆菌的GICA系统能够特异、快速检测杜克雷嗜血杆菌。该检测系统的灵敏度还不能满足所有临床标本的需要,有待于进一步研究。Objective To develop a rapid gold-immunochromatography assay （GICA） for detection of Haemophilus ducreyi. Method A sandwich GICA antigen detection system for H. ducreyi was developed by using the rabbit anti-rHgbAF peptide IgG as the capture antibody and the rabbit anti-rHgbA peptide IgG as the tracer antibody after being labeled with colloidal gold. By contrast with the ELISA system for H. ducreyi, the sensitivity and specificity of the test were evaluated using strains of H. ducreyi 35000, H. influenzae and other bacteria known to relate to genital ulcers. Results The results showed that H. ducreyi strain was strongly positive and all other bacteria were negative and the minimum amount of rHgbA detected was 6.25ng/mL （ELISA, 1.56ng/mL） and the minimum number of H. ducreyi detected was 2 × 10^5cfu/mL （ELISA, 2 ×10^5cfu/mL） in buffer and 1 × 10^6cfu/mL （ 1 × 10^6cfu/mL） in pus. The test could be finished in 15 min with a good stability. Conclusion Based on the specific polyclonal antibodies against rHgbA and rHgbAF, a rapid GICA system for H. ducreyi was developed primarily with a good specificity. Although the level of its sensitivity may not be sufficient to detect H. ducreyi in all clinical specimens, further work to increase the sensitivity could potentially make this assay a valuable tool in chancroid diagnosis.

关 键 词：杜克雷嗜血杆菌 软下疳 胶体金免疫层析技术 

分 类 号：R378.4[医药卫生—病原生物学]