New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies
New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies
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DOI：
10.1373/clinchem.2008.118299
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发表时间：
2009-05-01
影响因子：
9.3
通讯作者：
Komorowski, Lars
中科院分区：
文献类型：
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作者：
Daehnrich, Cornelia;Pares, Albert;Komorowski, Lars
背景技术背景：对原发性胆汁性肝硬化（PBC）特异性的抗线粒体抗体靶向2-含氧酸脱氢酶复合物的E2亚基，特别是丙酮酸脱氢酶复合物（PDC）-E2。它们的抗原特异性检测依赖于使用纯化的PDC的常规ELISA。最近的试验采用了含有3个E2亚基（MIT 3）的杂交体。一些PBC血清与一种或另一种制剂反应，表明存在非重叠epitopes.METHODS：我们已经开发了一种ELISA（抗M2 -3E），使用纯化的PDC和MIT 3的混合物作为抗原靶标。我们比较了这种检测抗MIT 3单独，传统的抗PDC，间接免疫荧光使用173 PBC和247疾病controls.RESULTS：抗M2 -3E ELISA显示了93.6%的诊断灵敏度相比，91.3%，83.8%，和87.3%的MIT 3，纯化的PDC，或间接免疫荧光，分别为98.8%时，所有的特异性设置为98.8%。用免疫印迹法检测，与纯化PDC不反应的抗M2 - 3E阳性血清可识别其它2种复合物的重组E2亚基，而与MIT 3免疫固定的PDC亚基E1 α a或E1 β不反应的抗M2 -3E阳性血清可识别其它2种复合物的重组E2亚基。结论：抗M2 -3E ELISA检测2-酮酸脱氢酶复合物抗体的准确性优于常规ELISA和IFL;其新奇来自于MIT 3杂交体和纯化PDC的组合。(C)2009年美国临床化学协会
BACKGROUND: Antimitochondrial antibodies specific for primary biliary cirrhosis (PBC) target the E2 Subunits of 2-oxo acid dehydrogenase complexes, in particular the pyruvate dehydrogenase complex (PDC)-E2. Their antigen-specific detection relies on conventional ELISA using purified PDC. More recent assays have employed a hybrid containing the 3 E2-subunits (MIT3). Some PBC sera react with one or the other preparation, suggesting the presence of nonoverlapping epitopes.METHODS: We have developed an ELISA (anti-M2-3E) using a mixture of purified PDC and MIT3 as antigenic targets. We compared this assay to anti-MIT3 alone, conventional anti-PDC, and indirect immunofluorescence using 173 PBC and 247 disease controls.RESULTS: The anti-M2-3E ELISA showed a 93.6% diagnostic sensitivity compared with 91.3%, 83.8%, and 87.3% for MIT3, purified PDC, or indirect immunofluorescence, respectively, when all specificities are set to 98.8%. By immunoblotting, anti-M2-3E-positive sera unreactive to purified PDC recognized recombinant E2-subunits of the other 2 complexes, whereas those with no reactivity to MIT3 immunofixed PDC subunits E1 alpha a or E1 beta.CONCLUSIONS: The diagnostic accuracy of the anti-M2-3E ELISA for detection of antibodies to 2-oxo acid dehydrogenase complexes exceeds that of conventional ELISA and IFL; its novelty derives from the combination of the MIT3 hybrid and purified PDC. (C) 2009 American Association for Clinical Chemistry