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光滑双脐螺和曼氏血吸虫共同抗原的分离、部分特性鉴定及其酶联免疫吸附测定和免疫印渍技术评估

Isolation and partial characterization of shared antigens of Biomphalaria glabrata and Schistosoma mansoni and their evaluation by the ELISA and the EITB.

作者信息

Rivera-Marrero C A, Hillyer G V

出版信息

J Parasitol. 1985 Oct;71(5):547-55.

PMID:3932633
Abstract

Serum from humans infected with Schistosoma mansoni, when reacted with a Biomphalaria glabrata soluble hepatopancreas antigen extract (BgSHA) yields 2 lines of precipitation by gel diffusion and 1 by immunoelectrophoresis. The IgG from the serum of a human infected with S. mansoni was coupled to CNBr-activated Sepharose 4B. BgSHA (8.0 mg) was then filtered through the gel and the bound antigens, denoted BgSm, eluted with HCl-glycine, pH 2.6. These bound antigens comprised 2.8% of the total BgSHA. BgSm was then applied to an anti-Fasciola hepatica column as above. The drop through in PBS (38% yield) and containing the BgSm antigens depleted of cross-reactivity with F. hepatica was then tested by the ELISA to evaluate its serodiagnostic potential. These antigens detected a primary S. mansoni infection by 4 wk but were less sensitive than SmSEA in the detection of a primary infection with S. mansoni. However, the BgSm-specific antigens were more specific than SmSEA and showed less cross-reactivity with the serum of mice infected with F. hepatica. At least 16 peptides were seen by silver staining following SDS-PAGE with 5-20% gradient gels. The 2 more prominent bands obtained were estimated to have molecular weights of 62 and 66 kd. Nitrocellulose strips blotted with BgSHA were incubated with the serum of mice infected with S. mansoni for 12 wk and developed 6 bands with molecular weights of 66, 57, 55, 50, 48 and 32 kd.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

感染曼氏血吸虫的人类血清与光滑双脐螺可溶性肝胰腺抗原提取物(BgSHA)反应时,通过凝胶扩散产生2条沉淀线,通过免疫电泳产生1条沉淀线。将感染曼氏血吸虫的人类血清中的IgG与溴化氰活化的琼脂糖4B偶联。然后将8.0毫克BgSHA通过该凝胶过滤,结合的抗原(称为BgSm)用pH 2.6的盐酸 - 甘氨酸洗脱。这些结合的抗原占总BgSHA的2.8%。然后将BgSm如上应用于抗肝片吸虫柱。然后通过ELISA测试在PBS中的流出液(产率38%),其含有与肝片吸虫交叉反应性已消除的BgSm抗原,以评估其血清学诊断潜力。这些抗原在4周时可检测到曼氏血吸虫的初次感染,但在检测曼氏血吸虫初次感染时不如曼氏血吸虫可溶性虫卵抗原(SmSEA)敏感。然而,BgSm特异性抗原比SmSEA更具特异性,并且与感染肝片吸虫的小鼠血清的交叉反应性更低。用5-20%梯度凝胶进行SDS-PAGE后,通过银染至少可见16种肽。获得的2条更明显的条带估计分子量为62和66kd。用BgSHA印迹的硝酸纤维素条与感染曼氏血吸虫12周的小鼠血清孵育,显露出分子量为66、57、55、50、48和32kd的6条带。(摘要截断于250字)

相似文献

1
Isolation and partial characterization of shared antigens of Biomphalaria glabrata and Schistosoma mansoni and their evaluation by the ELISA and the EITB.光滑双脐螺和曼氏血吸虫共同抗原的分离、部分特性鉴定及其酶联免疫吸附测定和免疫印渍技术评估
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Clin Diagn Lab Immunol. 2004 Nov;11(6):1165-70. doi: 10.1128/CDLI.11.6.1165-1170.2004.
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Schistosoma mansoni shares a protective carbohydrate epitope with keyhole limpet hemocyanin.曼氏血吸虫与钥孔戚血蓝蛋白共享一种保护性碳水化合物表位。
J Exp Med. 1987 Mar 1;165(3):865-78. doi: 10.1084/jem.165.3.865.