ELISA双抗原夹心法检测内脏利什曼病抗体的研究  

Study on the use of a double-antigen sandwich enzyme-linked immunosorbent assay (DAgS-ELISA) to detect antibodies to visceral Leishmaniasis

作　　者：雷刚[1] 徐恩洁[2] 吕天义[1] 徐艺玫[1] 张光朴 热娜.吐尔地[1] 窦海平 凯煞尔 徐秉臣[1] 廖力夫[1] 

机构地区：[1]新疆疾病预防控制中心,新疆乌鲁木齐830002 [2]新疆医科大学第一附属医院,新疆乌鲁木齐830001 [3]Rosalind Franklin大学芝加哥医学院微生物免疫,USA 60064 [4]喀什地区疾病预防控制中心,新疆喀什840002

出　　处：《中国病原生物学杂志》2010年第10期743-745,共3页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.30460120,30760217)

摘　　要：目的观察rK39抗原酶联免疫吸附试验双抗原夹心法(ELISA夹心法)检测内脏利什曼病抗体用于内脏利什曼病诊断与宿主动物感染调查的可行性。方法采用ELISA夹心法与rK39免疫层析试条法(ICT)同步检测内脏利什曼病抗体。结果 5种家畜血清229份,7种鼠类标本238份,ELISA夹心法和rK39 ICT法抗体检测均阴性;接种利什曼原虫灰仓鼠、草原兔尾鼠标本33份,13份阳性,阳性率39.4%;塔里木兔标本119份,阳性7份,阳性率5.9%;非疫区儿童血清29份,全部阴性;疫区无内脏利什曼病症状人血清250份,4份两种方法均阳性,阳性率均为1.6%;住院内脏利什曼病病人血清67份,两种方法的阳性率分别为68.7%和67.2%。共检测人和其他动物标本965份,两种方法的阳性符合率98.6%,阴性符合率99.9%。ICT相同显色等级的阳性标本,ELISA跨10个滴度。结论 ELISA夹心法可检测多种动物内脏利什曼病抗体,抗体滴度具有定量意义。该方法适用于内脏利什曼病诊断、疗效观察和流行病学调查。Objective To observe the feasibility of using an rK39 double-antigen sandwich enzyme-linked immunosorbent assay（DAgS-ELISA） to detect antibodies to visceral Leishmaniasis（VL） as a method to diagnose VL and investigate the infection of host animals.Methods Antibodies to VL were detected simultaneously with DAgS-ELISA and an rK39 immunochromatographic test（ICT）.Results Two hundred and twenty-nine serum samples from five kinds of domestic animals and 238 serum samples from seven kinds of rodents were negative according to DAgS-ELISA and the rK39 ICT.Of 33 samples from gray hamsters（Cricetulus migratorius） and steppe voles（Lagurus lagurus） inoculated with VL,13 were positive at a rate of 39.4%.Of 119 samples from Tarim hares（Lepus yarkandensis）,7 were positive at a rate of 5.8%.All samples from 29 children in areas where the disease was not prevalent were negative.Of 250 samples from people with no symptoms of VL in areas where VL was prevalent,4 were positive at a rate of 1.6%.Of 67 samples from hospitalized patients with VL,samples were positive at a rate of 68.7% and 67.2%,respectively,according to DAgS-ELISA and rK39 ICT.Nine hundred and sixty-five samples from humans and a variety of animals were tested,and the rate of concordance for a positive reaction according to the two methods was 98.6% and the corresponding rate of concordance for a negative reaction was 99.9%.Positive samples with equivalent coloration according to ICT had a titer 10-fold that according to DAgS-ELISA.Conclusion Antibodies to VL were detected by DAgS-ELISA in a variety of animals and antibody titers were quantitative.DAgS-ELISA can be used to diagnose VL and assist in assessing efficacy and conducting epidemiological studies.

关 键 词：内脏利什曼病 rK39 抗体 酶联免疫吸附试验双抗原夹心法 

分 类 号：R531.6[医药卫生—内科学]