摘要
目的：建立一种检测感染宿主尿内排泄抗原的简易免疫学试验。方法：用经改良的兼具ｄｏｔ－ＥＬＩＳＡ和酶标板－ＥＬＩＳＡ优点的试剂条双夹心酶联免疫吸附试验，检测了家兔及人宿主尿样本中的日本血吸虫肠相关趋阴极循环抗原（ＣＣＡ）。结果：１０只高感染兔尿样经透析冻干保存达１０年，复原后检测均呈强阳性反应，滴度可达５１２－１，１２只经相同处理的正常兔尿，仅１只出现８－１以下的弱反应。５８例采自四川省的感染者和６０例正常对照者尿液的ＣＣＡ检测，敏感性和特异性分别为４３．１％和９６．７％。与板－ＥＬＩＳＡ比较，检出率相仿而呈现一定的互补效应，互补检出率为５８．６％。兔尿样内加入适量捕获单抗，反应强度即出现剂量依赖的竞争抑制，提示了尿内被检测到的靶微粒具有被抗ＣＣＡ单抗特异地识别的属性。对于采用同一检测系统不同方法的互补检测性也进行了讨论。结论：实验结果不仅显示了试剂条夹心ＥＬＩＳＡ适合现场应用的诸多优点，也证明用于特异地检测感染宿主尿内排泄抗原的可行性。
AIM:To set up a simple immunoassay for detection of the excreted antigens in host urine.METHOD:A modified double anitbody sandwich ELISA in the form of a strip reagent which combined the advantages of both dot ELISA and plate ELISA for the detection of the gut associated circulating cathodic antigen(CCA) in the urine of Schistosoma japonicum infected rabbits and patients was developed.RESULTS:All the 9 reconstituted lyophilized urine samples collected from heavily infected rabbits,stored for over 10 years,gave strong positive reactions up to an end point titre of 512 -1 ,while only one out of 11 urine samples from normal rabbit stored and reconstituted in the same manner showed a weak reaction which faded at a dilution of 8 -1 .Urine CCA detections in 58 schistosomiasis patients collected from Sichuan Province,China and in 60 healthy students from the Shanghai Second Medical University revealed an assay sensitivity of 43.1% and a specificity of 96.7%. The detectability has been found similar to the parallelly performed plate ELISA for urine CCA.A certain degree of diagnostic complementarity in parallel up to 58.6% in these patient samples was seen with no increase of false positive cases.For identification of the diagnostic specificity,an appropriate amount of the capture antibody was added to the infected rabbit urine,a dose dependent competitive inhibition of the reaction intensity was observed,indicating the specific recognition of the target molecules in the urine by the anti CCA McAb system.The possible diagnostic complementarity via different methods using the same probe system was discussed.CONCLUSION:Results of this study have revealed the possibility as well as various advantages of the sandwich reagent strip assay to be used in the field for an non invasive detection of the excreted antigens in urine of infected individuals.
出处
《中国寄生虫学与寄生虫病杂志》
CSCD
北大核心
1996年第4期253-256,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
卫生部及欧共体STD-3项目的联合资助
关键词
日本血吸虫
尿抗原
检测
免疫试验
Schistosoma japonicum
urine antigen detections
urine CCA
strip reagent immunoassay
