Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay

Danilo Villalta, Maria Concetta Sorrentino, Elia Girolami, Marilina Tampoia, Maria Grazia Alessio, Ignazio Brusca, Massimo Daves, Brunetta Porcelli, Giuseppina Barberio, Nicola Bizzaro

Research output: Contribution to journalArticle

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Abstract

Objective: To evaluate the autoantibody profile in patients with primary biliary cirrhosis (PBC) using a new multiplexed line-blot assay specifically designed for the diagnosis of autoimmune liver diseases. Methods: Sera of 58 consecutive PBC patients and 191 disease controls (144 with autoimmune liver diseases other than PBC, and 67 with non-autoimmune chronic liver diseases) were tested by both the multiplexed line-blot Autoimmune Liver Disease Profile 2 (ALD2) and by IIF on HEp-2 cells and on rat kidney/liver/stomach tissues. ALD2 contains the following PBC-associated antigens: AMA-M2, natively purified from bovine heart; M2-E3, a recombinant fusion protein including the E2 subunits of PDC, BCOADC and OGDC; sp100, PML and gp210 recombinant proteins. Results: With the ALD2 assay, a positive reaction to AMA-M2, M2-E3, sp100, PML and gp210 in PBC patients was observed in 77.6%, 84.5%, 34.5%, 15.1% and 18.9%, respectively, of the PBC sera. The overall sensitivity and specificity for PBC were 98.3% and 93.7%. Using IIF, positivity rates to AMA, and to antinuclear autoantibodies with membranous/rim-like and multiple nuclear dot patterns were 86.2%, 8.6% and 22.4%, respectively. The overall sensitivity and specificity for PBC of the IIF method were 86.2% and 97.9%, respectively. Conclusions: The ALD2 line-blot showed a good diagnostic accuracy for PBC and a higher sensitivity than the IIF method to detect sp100 and gp210 autoantibodies.

Original languageEnglish
Pages (from-to)135-138
Number of pages4
JournalClinica Chimica Acta
Volume438
DOIs
Publication statusPublished - Jan 1 2015

Fingerprint

Biliary Liver Cirrhosis
Liver
Autoantibodies
Assays
Liver Diseases
Autoimmune Diseases
Disease control
Recombinant Fusion Proteins
Sensitivity and Specificity
Recombinant Proteins
Serum
Rats
Stomach
Chronic Disease
Tissue
Antigens
Kidney

Keywords

  • Anti-mitochondrial autoantibodies (AMA)
  • Autoantibodies
  • Gp210
  • Membranous/rim-like
  • Multiple nuclear dots (MND)
  • Sp100

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical
  • Medicine(all)

Cite this

Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay. / Villalta, Danilo; Sorrentino, Maria Concetta; Girolami, Elia; Tampoia, Marilina; Alessio, Maria Grazia; Brusca, Ignazio; Daves, Massimo; Porcelli, Brunetta; Barberio, Giuseppina; Bizzaro, Nicola.

In: Clinica Chimica Acta, Vol. 438, 01.01.2015, p. 135-138.

Research output: Contribution to journalArticle

Villalta, D, Sorrentino, MC, Girolami, E, Tampoia, M, Alessio, MG, Brusca, I, Daves, M, Porcelli, B, Barberio, G & Bizzaro, N 2015, 'Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay', Clinica Chimica Acta, vol. 438, pp. 135-138. https://doi.org/10.1016/j.cca.2014.08.024
Villalta, Danilo ; Sorrentino, Maria Concetta ; Girolami, Elia ; Tampoia, Marilina ; Alessio, Maria Grazia ; Brusca, Ignazio ; Daves, Massimo ; Porcelli, Brunetta ; Barberio, Giuseppina ; Bizzaro, Nicola. / Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay. In: Clinica Chimica Acta. 2015 ; Vol. 438. pp. 135-138.
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abstract = "Objective: To evaluate the autoantibody profile in patients with primary biliary cirrhosis (PBC) using a new multiplexed line-blot assay specifically designed for the diagnosis of autoimmune liver diseases. Methods: Sera of 58 consecutive PBC patients and 191 disease controls (144 with autoimmune liver diseases other than PBC, and 67 with non-autoimmune chronic liver diseases) were tested by both the multiplexed line-blot Autoimmune Liver Disease Profile 2 (ALD2) and by IIF on HEp-2 cells and on rat kidney/liver/stomach tissues. ALD2 contains the following PBC-associated antigens: AMA-M2, natively purified from bovine heart; M2-E3, a recombinant fusion protein including the E2 subunits of PDC, BCOADC and OGDC; sp100, PML and gp210 recombinant proteins. Results: With the ALD2 assay, a positive reaction to AMA-M2, M2-E3, sp100, PML and gp210 in PBC patients was observed in 77.6{\%}, 84.5{\%}, 34.5{\%}, 15.1{\%} and 18.9{\%}, respectively, of the PBC sera. The overall sensitivity and specificity for PBC were 98.3{\%} and 93.7{\%}. Using IIF, positivity rates to AMA, and to antinuclear autoantibodies with membranous/rim-like and multiple nuclear dot patterns were 86.2{\%}, 8.6{\%} and 22.4{\%}, respectively. The overall sensitivity and specificity for PBC of the IIF method were 86.2{\%} and 97.9{\%}, respectively. Conclusions: The ALD2 line-blot showed a good diagnostic accuracy for PBC and a higher sensitivity than the IIF method to detect sp100 and gp210 autoantibodies.",
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AU - Villalta, Danilo

AU - Sorrentino, Maria Concetta

AU - Girolami, Elia

AU - Tampoia, Marilina

AU - Alessio, Maria Grazia

AU - Brusca, Ignazio

AU - Daves, Massimo

AU - Porcelli, Brunetta

AU - Barberio, Giuseppina

AU - Bizzaro, Nicola

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N2 - Objective: To evaluate the autoantibody profile in patients with primary biliary cirrhosis (PBC) using a new multiplexed line-blot assay specifically designed for the diagnosis of autoimmune liver diseases. Methods: Sera of 58 consecutive PBC patients and 191 disease controls (144 with autoimmune liver diseases other than PBC, and 67 with non-autoimmune chronic liver diseases) were tested by both the multiplexed line-blot Autoimmune Liver Disease Profile 2 (ALD2) and by IIF on HEp-2 cells and on rat kidney/liver/stomach tissues. ALD2 contains the following PBC-associated antigens: AMA-M2, natively purified from bovine heart; M2-E3, a recombinant fusion protein including the E2 subunits of PDC, BCOADC and OGDC; sp100, PML and gp210 recombinant proteins. Results: With the ALD2 assay, a positive reaction to AMA-M2, M2-E3, sp100, PML and gp210 in PBC patients was observed in 77.6%, 84.5%, 34.5%, 15.1% and 18.9%, respectively, of the PBC sera. The overall sensitivity and specificity for PBC were 98.3% and 93.7%. Using IIF, positivity rates to AMA, and to antinuclear autoantibodies with membranous/rim-like and multiple nuclear dot patterns were 86.2%, 8.6% and 22.4%, respectively. The overall sensitivity and specificity for PBC of the IIF method were 86.2% and 97.9%, respectively. Conclusions: The ALD2 line-blot showed a good diagnostic accuracy for PBC and a higher sensitivity than the IIF method to detect sp100 and gp210 autoantibodies.

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KW - Anti-mitochondrial autoantibodies (AMA)

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KW - Membranous/rim-like

KW - Multiple nuclear dots (MND)

KW - Sp100

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