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［摘 要］ 目的 制备抗沙眼衣原体（CT）沙眼生物变种的特异性单克隆抗体(mAb)，并建立CT的胶体金免疫层析快速检测方法。
方法 CT脂多糖(LPS)免疫BALB/C小鼠，通过细胞融合，建立分泌抗CT抗体的杂交瘤细胞株，用ELISA法鉴定mAb的免疫球蛋白亚类和与衣原体性病淋巴肉芽肿生物变种（CLGV）的交叉反应。用中和抑制法测定mAb与w1型淋球菌、w2型淋球菌、人型支原体、解脲支原体的交叉反应；配对试验挑选出两株mAb，建立检测CT的双抗体夹心法免疫层析试纸条，并与进口胶体金检测卡对照检测176份女性宫颈和男性尿道拭子标本。
结果 配对较好的8株mAb中6株为小鼠IgG1，1株为IgG2a，1株为IgM，8株mAb与CLGV、w1型淋球菌、w2型淋球菌、人型支原体、解脲支原体均无交叉反应。与进口胶体金检测卡相比，阴性符合率为98.65％，阳性符合率为89.29%，总符合率为97.16％。
结论 本研究制备的mAb具有较高的特异性，建立的检测方法可应用于CT引起的泌尿生殖道感染的检测。
［关键词］ 沙眼衣原体;单克隆抗体;胶体金;免疫层析
［中图分类号］
R 374.1
［文献标识码］ B
［文章编号］ 1001-7089(2004)05-0230-02
Preparation
and Application of Monoclonal Antibody Against C.Trachomatis
YANG
Shu-hao,ZHENG Peng-ju,WANG yu-zhu,et
al
(Zhengzhou Biocell Institute,Zhengzhou 450001,China)
Abstract:
Objective To prepare
monoclonal antibodies(mAb) against C. trachomatis(CT) and establish a new
method to detect CT.
Methods
Hybridoma lines were obtained by fusing myeloma SP2/0 and
spleen cells from BALB/c mouse immuned with lipopolysaccharide(LPS) of
CT.Isotype of the mAbs was determined by ELISA.Specificity of the mAbs was
identified by the method of ELISA and inhibition ELISA.Two mAbs were
selected to make up colloidal gold immunochromatographic assay(CIA) strip
for testing CT antigen.This method was compared with CLEARVIEW test card by
testing 176 cervical(female) and urethral(male) smears samples.
Results
The isotypes of 6 out of 8 monoclonal antibodies were IgG1
while the other two were IgG2a and IgM,respectively.8 monoclonal
antibodies didn’t react with chamydia biovar
lymphogranuloma venereum(CLGV),w1 type N.gonorrhoeae,w2
type N.gonorrhoeae,M.hominis,and U.urealyticum.Compared with CLEARVIEW test
card,the sensitivity,specificity and overall agreement of CIA strip were
98.65％,94.74% and 97.31％,respectively.
Conclusion
2 mcAbs have high specificity.CIA strip made up of these
mAb can be applied to detect CT for the cervical and urethral smears
samples.
Key words：
C.trachomatis;Monoclonal antibodies;Colloidal
gold;Immunochromatographic assay
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