自制免疫胶体金层析条检测恶性疟原虫的初步研究  

Diagnosis of Plasmodium falciparum by gold-immunochromatography assay

作　　者：吴英松[1] 李明[1] 毕惠祥[1] 董文其[1] 李英杰[1] 

机构地区：[1]第一军医大学热带病研究室,广州510515

出　　处：《中华微生物学和免疫学杂志》2002年第2期229-232,共4页Chinese Journal of Microbiology and Immunology

基　　金：全军医药卫生科研基金课题 ( 96L0 34)

摘　　要：目的　建立一种简易快速、适合于基层使用的自测式免疫胶体金层析 (GICA)法用于恶性疟原虫的检测。方法　采用柠檬酸三钠还原法制备胶体金颗粒 ,标记抗恶性疟原虫乳酸脱氢酶(LDHpf)单抗 2A5 ,并以另外 4株单抗作为包被抗体 ,制成诊断恶性疟原虫的免疫层析条 ,探索最佳配对模式及估计最低检测量。以镜检和PCR法为对照 ,用GICA条检测门诊“四热”病人血样 ,评价其敏感性和特异性 ,并对其稳定性进行了初步研究。结果　GICA检测重组LDHpf,最低检测量为 1ng。以镜检法和PCR法为标准 ,GICA条检测恶性疟原虫的敏感性分别为 88.37%和 86 .6 7% ,GICA与镜检法的符合率均为 91.5 5 %。并且当虫体密度 >2 0 0个 μl时 ,GICA的敏感性为 10 0 %。GICA条在 4℃下可保存 3个月以上。结论　GICA检测恶性疟原虫简易快速、灵敏度高 ,无需特殊仪器设备 。Objective To establish a simple, fast and selfperforming Gold-immunochromatography assay(GICA) for detecting Plasmodium falciparum. Methods McAbs against lactate dehydrogenase of P. falciparum (LDHpf) were purified and then applied to preparation of GICA strips. With microscopic examination of blood smears and PCR test as control, GICA was used to detect malaria in outpatient clinics of China. Its sensitivity, specificity and stability were evaluated. Results Threshold value of GICA was 1 ng by detecting the recombinant LDHpf antigen. Compared with micro scopy of blood smears and PCR test, GICA showed sensitivities of 88.37% and 86.67% respectively. Concordance between the GICA and microscopic examination for P. falciparum was 94.37%. GICA test showed 100% sensitivity at the infection levels >200parasites/μl, but sensitivity decreased markedly at the level <200parasites/μl. GICA strips coated with antigen and antibody can be stored at 4℃ for more than three months. Conclusion GICA established in this study is simple, fast, sensitive and specific for detecting P. falciparum, and is potentially useful in developing diagnostic kits.

关 键 词：免疫胶体金层析 诊断 恶性疟原虫 

分 类 号：R446.6[医药卫生—诊断学]