Chomvarin C, Chantarasuk Y, Thongkrajai P, Yutanawiboonchai W, Waropastrakul N
Department of Microbiology, Faculty of Medicine, Khon Kaen University, Thailand.
Southeast Asian J Trop Med Public Health. 2000 Mar;31(1):96-103.
A double antibody sandwich enzyme immunoassay (EIA) for chlamydial antigen detection was developed using a monoclonal antibody against lipopolysaccharide (LPS) of Chlamydia trachomatis as a coating antibody. Polyclonal rabbit antiserum against partially purified antigen from elementary body (EB) antibody and horse-radish peroxidase conjugated goat anti-rabbit antibody were used as the primary and secondary antibody respectively. The developed EIA could detect protein of partially purified EB at the lowest concentration of 250 ng/ml. The assay was evaluated against the cell culture (CC), DNA hybridization assay (PACE2 system: Gen-Probe, San Diego, CA, USA) and a commercial enzyme immunoassay (kEIA) (Bioquest, NSW, Australia). The sensitivity, specificity, positive and negative predictive values of the developed EIA (dEIA) were 87, 96.2, 80, 97.7 for the specimens from females and 90.9, 90.7, 71.4, 97.5 for the specimens from males repectively. Cross reaction was not found with Escherichia coli, Acinetobacter anitratus, beta-Streptococcus group A, Enterobacter spp, Enterococcus, Lactobacillus spp, Neisseria spp, but it was found with Candida albicans and herpes simplex virus type 1. The developed EIA can be applied successfully for both genders, particularly males. The cost per test is less than those for CC, kEIA and PACE2.
开发了一种用于检测衣原体抗原的双抗体夹心酶免疫测定法（EIA），使用针对沙眼衣原体脂多糖（LPS）的单克隆抗体作为包被抗体。分别使用针对原体（EB）抗体部分纯化抗原的兔抗血清和辣根过氧化物酶标记的山羊抗兔抗体作为一抗和二抗。所开发的EIA能够检测最低浓度为250 ng/ml的部分纯化EB的蛋白质。该测定法与细胞培养（CC）、DNA杂交测定法（PACE2系统：Gen-Probe，美国加利福尼亚州圣地亚哥）和一种商业酶免疫测定法（kEIA）（Bioquest，澳大利亚新南威尔士州）进行了评估。所开发的EIA（dEIA）对于女性标本的敏感性、特异性、阳性和阴性预测值分别为87、96.2、80、97.7，对于男性标本分别为90.9、90.7、71.4、97.5。未发现与大肠杆菌、鲍曼不动杆菌、A组β溶血性链球菌、肠杆菌属、肠球菌、乳酸杆菌属、奈瑟菌属发生交叉反应，但与白色念珠菌和1型单纯疱疹病毒发生了交叉反应。所开发的EIA可成功应用于男性和女性，尤其是男性。每次检测的成本低于CC、kEIA和PACE2。
Zotero 插件