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葡萄球菌GST-mTSST-1融合蛋白抗体间接ELISA
检测方法的建立
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郭海勇1，3，崔京春2，钱爱东3
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（1.吉林师范大学 生命科学学院，吉林 四平136000； 2.大连民族学院 生命科学学院，辽宁 大连
116600；3.吉林农业大学 动物科学技术学院,吉林 长春130118)
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摘要：
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以纯化的金黄色葡萄球菌无毒诱变超抗原GST-mTSST-1融合蛋白作为检测抗原，通过优化ELISA反应条件，初步建立了检测血清特异性GST-mTSST-1抗体的间接ELISA方法，监测了小鼠血清中特异性GST-mTSST-1抗体水平的动态变化规律。方阵试验确定的GST-mTSST-1抗原的最适包被浓度为5.0μg/mL，血清最佳稀释倍数为1∶100，ELISA阳性反应的临界值为D490nm≥0.219，批内和批间重复试验的变异系数均小于10%。应用该方法对试验小鼠血清进行检测，结果表明,建立的间接ELISA方法具有较高的敏感性、特异性和较好的重复性，适用于检测特异性GST-mTSST-1血清抗体。 |
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关键词：
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金黄色葡萄球菌；GST-mTSST-1；融合蛋白；间接ELISA；超抗原 |
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中图分类号：S 852.611：R 446.61 文献标识码：
A 文章编号：1673-4696(2007)12-1041-05 |
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Development of an indirect ELISA assay for detecting antibody against
Staphylococcus aureus GST-mTSST-1 fusion protein |
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GUO Hai-yong1,3， CUI Jing-chun2，QIAN
Ai-dong3 |
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(1.College of Life Sciences,Jilin Normal University,Siping 136000,China；2.College of Life Sciences,
Dalian Nationalities University,Dalian 116600,China；3.College of Animal Science and Technology,
Jilin Agricultural University,Changchun 130118,China) |
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Abstract: Using the purified nontoxic and mutant superantigen glutathione S-transferase and mutation toxic shock syndrome toxic-1(GST-mTSST-1) fusion protein of Staphylococcus aureus as coating antigen,an indirect ELISA assay was developed for the detection of sera antibody against specific TSST-1 toxin by optimizing the reaction conditions.In the indirect ELISA,the optimal concentration of coating antigen was 5.0μg/mL and the optimal dilution of serum sample was 1∶100,the cutoff value for positive response was D490nm≥0.219.In the intro-batch and the inter-batch repeated test,the variation coefficient was less than 10%.The murine serum samples were detected by the indirect ELISA to monitor the dynamic changes and the regularity of antibody titer in serum.The results showed that the indirect ELISA assay was of high sensitiveness,good reproducibility and specificity,suitable to detect the specific anti-serum for GST-mTSST-1 of S.aureus.
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Key words: Staphylococcus aureus； glutathione S-transferase and mutation toxic shock syndrome toxic-1；fusion protein；indirect enzyme-linked immunosorbent assay；superantigen |
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