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使用与曼氏血吸虫15千道尔顿被膜抗原反应的单克隆抗体进行快速竞争性酶联免疫吸附测定以用于血吸虫病的血清学诊断。

Rapid competitive enzyme-linked immunosorbent assay using a monoclonal antibody reacting with a 15-kilodalton tegumental antigen of Schistosoma mansoni for serodiagnosis of schistosomiasis.

作者信息

Da Silva A J, Piuvezam M R, de Moura H, Maddison S, Peralta J M

机构信息

Centers for Disease Control, Atlanta, Georgia 30341-3724.

出版信息

J Clin Microbiol. 1993 Sep;31(9):2315-9. doi: 10.1128/jcm.31.9.2315-2319.1993.

Abstract

A competitive enzyme-linked immunosorbent assay (CELISA) for antibody detection was developed by using a monoclonal antibody which reacts with a 15-kDa tegumental antigen of the adult worm of Schistosoma mansoni. This monoclonal antibody was not able to react with antigens of Schistosoma japonicum or Schistosoma haematobium in enzyme-linked immunoelectrotransfer blot (EITB) and indirect immunofluorescence tests. The assay was performed in a period of 1 h using an adult worm crude extract antigen. To evaluate the CELISA, a total of 73 serum samples was analyzed: 35 were from S. mansoni-infected patients, 23 were from individuals with parasitic infections other than schistosomiasis, and 14 were from healthy individuals. All serum samples from healthy individuals and from patients infected with other parasites were negative, as were two (6%) samples from patients infected with S. mansoni. EITB analysis showed that 32 of 33 CELISA-positive samples were positive in the EITB but with different patterns of reactivity. A 15-kDa protein reacted with 60% of serum samples, and a 60-kDa protein showed the highest level of reactivity (85%). The two samples from patients infected with S. mansoni that were negative in the CELISA reacted with 70-, 60-, 50-, 47-, and 38-kDa proteins. One sample, positive in CELISA, did not react with proteins of the antigenic extract.

摘要

利用一种单克隆抗体开发了一种用于抗体检测的竞争性酶联免疫吸附测定(CELISA),该单克隆抗体可与曼氏血吸虫成虫的一种15 kDa皮层抗原发生反应。在酶联免疫电转移印迹(EITB)和间接免疫荧光试验中,这种单克隆抗体不能与日本血吸虫或埃及血吸虫的抗原发生反应。该测定使用成虫粗提物抗原在1小时内完成。为了评估CELISA,共分析了73份血清样本:35份来自曼氏血吸虫感染患者,23份来自除血吸虫病外患有寄生虫感染的个体,14份来自健康个体。所有健康个体和感染其他寄生虫患者的血清样本均为阴性,曼氏血吸虫感染患者的两份样本(6%)也为阴性。EITB分析显示,33份CELISA阳性样本中的32份在EITB中呈阳性,但反应模式不同。一种15 kDa蛋白与60%的血清样本发生反应,一种60 kDa蛋白显示出最高的反应水平(85%)。CELISA检测呈阴性的两名曼氏血吸虫感染患者的样本与70、60、50、47和38 kDa的蛋白发生反应。一份CELISA检测呈阳性的样本未与抗原提取物的蛋白发生反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4793/265753/229e5344292b/jcm00021-0083-a.jpg

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