New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies

Cornelia Daehnrich; Albert Pares; Llorenc Caballeria; Anke Rosemann; Wolfgang Schlumberger; Christian Probst; Maria Mytilinaiou; Dimitrios Bogdanos; Diego Vergani; Winfried Stoecker; Lars Komorowski

doi:10.1373/clinchem.2008.118299

New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies

Cornelia Daehnrich, Albert Pares, Llorenc Caballeria, Anke Rosemann, Wolfgang Schlumberger, Christian Probst, Maria Mytilinaiou, Dimitrios Bogdanos, Diego Vergani, Winfried Stoecker, Lars Komorowski

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Abstract

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

Original language	English
Pages (from-to)	978 - 985
Number of pages	8
Journal	Clinical Chemistry
Volume	55
Issue number	5
DOIs	https://doi.org/10.1373/clinchem.2008.118299
Publication status	Published - 1 May 2009

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10.1373/clinchem.2008.118299

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title = "New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies",

abstract = "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",

author = "Cornelia Daehnrich and Albert Pares and Llorenc Caballeria and Anke Rosemann and Wolfgang Schlumberger and Christian Probst and Maria Mytilinaiou and Dimitrios Bogdanos and Diego Vergani and Winfried Stoecker and Lars Komorowski",

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doi = "10.1373/clinchem.2008.118299",

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T1 - New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies

AU - Daehnrich, Cornelia

AU - Pares, Albert

AU - Caballeria, Llorenc

AU - Rosemann, Anke

AU - Schlumberger, Wolfgang

AU - Probst, Christian

AU - Mytilinaiou, Maria

AU - Bogdanos, Dimitrios

AU - Vergani, Diego

AU - Stoecker, Winfried

AU - Komorowski, Lars

PY - 2009/5/1

Y1 - 2009/5/1

N2 - 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

AB - 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

U2 - 10.1373/clinchem.2008.118299

DO - 10.1373/clinchem.2008.118299

M3 - Article

VL - 55

SP - 978

EP - 985

JO - Clinical Chemistry

JF - Clinical Chemistry

IS - 5

ER -

New ELISA for Detecting Primary Biliary Cirrhosis-Specific Antimitochondrial Antibodies

Abstract

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