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通过对低传播地区患者血清进行免疫印迹分析检测曼氏血吸虫膜抗原

Detection of Schistosoma mansoni membrane antigens by immunoblot analysis of sera of patients from low-transmission areas.

作者信息

Cesari Italo M, Ballen Diana E, Mendoza Leydi, Matos César

机构信息

Laboratorio de Inmunoparasitología, Centro de Microbiología y Biología Celular, Instituto Venezolano de Investigaciones Científicas, Apdo. 21827, Caracas 1020 A, Venezuela.

出版信息

Clin Diagn Lab Immunol. 2005 Feb;12(2):280-6. doi: 10.1128/CDLI.12.2.280-286.2005.

DOI:10.1128/CDLI.12.2.280-286.2005
PMID:15699423
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC549304/
Abstract

Schistosoma mansoni surface membrane components play a relevant role in the host-parasite interaction, and some are released in vivo as circulating antigens. n-Butanol extraction favors the release of membrane antigens like alkaline phosphatase, which has been shown to be specifically recognized by antibodies from S. mansoni-infected humans and animals. In the present study, components in the n-butanol extract (BE) of the adult S. mansoni worm membrane fraction were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D SDS-PAGE [15%]) and further analyzed by immunoblotting (immunoglobulin G) using defined sera. S. mansoni-infected patient sera, but not sera of uninfected patients or sera obtained from patients infected with other parasite species, specifically and variably recognized up to 20 polypeptides in the molecular mass range of approximately 8 to >80 kDa. There were some differences in the number, intensity, and frequency of recognition of the BE antigens among sera from Venezuelan sites of endemicity with a different status of schistosomiasis transmission. Antigens in the 28- to 24-kDa molecular mass range appeared as immunodominants and were recognized by S. mansoni-positive sera from all the sites, with recognition frequencies varying between 57.5 and 97.5%. Immunoblotting with BE membrane antigens resulted in a highly sensitive (98.1%), specific (96.1.0%), and confirmatory test for the immunodiagnosis of schistosomiasis in low-transmission areas.

摘要

曼氏血吸虫表面膜成分在宿主 - 寄生虫相互作用中发挥着重要作用,其中一些成分作为循环抗原在体内释放。正丁醇提取有利于膜抗原如碱性磷酸酶的释放,碱性磷酸酶已被证明能被曼氏血吸虫感染的人和动物的抗体特异性识别。在本研究中,成年曼氏血吸虫虫体膜部分的正丁醇提取物(BE)中的成分通过一维十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(1D SDS - PAGE [15%])进行分离,并使用特定血清通过免疫印迹法(免疫球蛋白G)进一步分析。曼氏血吸虫感染患者的血清,而非未感染患者的血清或感染其他寄生虫物种患者的血清,能特异性且不同程度地识别分子量范围约为8至>80 kDa的多达20种多肽。在委内瑞拉不同血吸虫病传播状态的流行地区的血清中,对BE抗原的识别数量、强度和频率存在一些差异。分子量在28至24 kDa范围内的抗原表现为免疫显性抗原,所有地点的曼氏血吸虫阳性血清均能识别,识别频率在57.5%至97.5%之间变化。用BE膜抗原进行免疫印迹对低传播地区血吸虫病的免疫诊断具有高度敏感性(98.1%)、特异性(96.1.0%)且具有确诊性。

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