摘要
目的 建立快速、简便地检测内脏利什曼病特异抗体的胶体金免疫层析试条方法,并评价效果. 方法 采用柠檬酸三钠还原法制备胶体金,用以标记链球菌G蛋白（streptococcal protein G,SPG）,并将其吸附于交联释放垫上;将杜氏利什曼原虫前鞭毛体粗抗原作为包被抗原,包被于硝酸纤维素膜适当位置,制成检测特异抗体的免疫层析试条.用该试条检测病原学确诊的内脏利什曼病（129例）、疟疾（20例）、细粒棘球蚴病（10例）、日本血吸虫病（10例）、并殖吸虫病（5例）、华支睾吸虫病（5例）等患者血清,以及健康者（40例）血清,评价其敏感性和特异性.同时用酶联免疫吸附试验（enzyme-linked immunosorbent assay,ELISA）进行平行检测. 结果 试条法检测129份黑热病患者血清,124份为阳性,敏感性为96.1％;与疟疾患者血清存在10.0％ （2/20）的交叉反应;与日本血吸虫病患者血清存在10.0％ （1/10）的交叉反应;与健康者血清存在2.5％ （1/40）的假阳性反应;与10份细粒棘球蚴病患者血清,5份并殖吸虫病患者血清和5份华支睾吸虫病患者血清均无交叉反应,总特异性为95.6％ （86/90）.该方法与ELISA法的符合率为99.2％,二者阳性检出率之间的差异无统计学意义（x2=0.12,P＞0.05）,二者特异性之间的差异也无统计学意义（x2=0.42,P＞0.05）. 结论 成功建立快速检测内脏利什曼病的胶体金免疫层析试条,该试条敏感性、特异性均较高.
Objective To establish and evaluate a colloid gold immunochromatographic strip test for detection of visceral leishmaniasis.Methods The streptococcal protein G（SPG） was conjugated with colloid gold as detecting reagent,visceral leishmaniasis antigen and anti-SPG IgY were immobilized on nitrocellulose at proper position,respectively.Serum samples of 129 visceral leishmaniasis patients were used for evaluating the sensitivity.Serum samples from patients with malaria（20 cases）,cystechinococcosis （10 cases）,schistosomiasis japonica（10 cases）,paragonimiasis （5 cases）,clonorchiosis（5 cases） and 40 healthy people were used for evaluating the specificity.Detection results of the immunochromatographic strip were compared with that of ELISA.Results Detection on 129 serum samples of patients with visceral leishmaniasis showed a sensitivity of 96.1％（124/129）.The cross-reactivity to malaria and schistosomiasis was 10.0％（2/20） and 10.0％ （1/10）,respectively.There was no cross-reactivity with schistosomiasis japonica,paragonimiasis and clonorchiosis.One sample out of 40 healthy people showed false positive reaction.The overall specificity was 95.6％（86/90）.The corresponding rate of immunochromatographic strip test with ELISA was 99.2％.There was no statistical difference in the sensitivity between immunochromatographic strip test and ELISA （x2 =0.12,P ＞ 0.05）.Specificity showed no statistical difference between immunochromatographic strip test and ELISA （x2 =0.42,P＞ 0.05）.Conclusion The immunochromatographic strip test was established succesfully,and is a sensitive,specific,simple and rapid assay for visceral leishmaniasis detection.
出处
《国际医学寄生虫病杂志》
CAS
2014年第3期121-124,共4页
International JOurnal of Medical Parasitic Diseases
关键词
内脏利什曼病
免疫层析试条
检测
评价
Visceral leishmaniasis
Immunochromatographic strip
Detection
Evaluation
