Clinical Rheumatology

, Volume 31, Issue 3, pp 503–509

A novel automated indirect immunofluorescence autoantibody evaluation

  • Shaye Kivity
  • Boris Gilburd
  • Nancy Agmon-Levin
  • Marina Garcia Carrasco
  • Yaron Tzafrir
  • Yael Sofer
  • Matilda Mandel
  • Thomas Buttner
  • Dirk Roggenbuck
  • Marco Matucci-Cerinic
  • Katalin Danko
  • Marcos López Hoyos
  • Yehuda Shoenfeld
Original Article

DOI: 10.1007/s10067-011-1884-1

Cite this article as:
Kivity, S., Gilburd, B., Agmon-Levin, N. et al. Clin Rheumatol (2012) 31: 503. doi:10.1007/s10067-011-1884-1

Abstract

Autoantibodies (AAb), especially antinuclear (ANAs) and anticytoplasmatic antibodies (ACyA), are essential diagnosing markers for several autoimmune diseases. The current gold standard method for ANA detection is manual indirect immunofluorescence (IIF) on human epithelial-2 (HEp-2) cells. However, this technique is cost and time consuming, and characterized by considerable intra- and interlaboratory variability. Thus, an automated IIF-HEp-2 reader has been developed recently. In the current study, we compared the performance of the automated AAb IIF-HEp-2 interpretation to conventional detection methods. Autoantibody detection by IIF on HEp-2 cells was performed in a total of 260 sera of patients, including 34 with systemic lupus erythematosus, 111 with dermatomyositis or polymyositis, 74 with systemic sclerosis, 41 with rare AAb patterns, and 137 healthy individuals. Visual interpretation and routine immunoassays were compared with a novel automated IIF-HEp-2 system using Aklides pattern recognition algorithms. Positive AAbs were detected in 95–100% of rheumatic patients by automated interpretation, in 74–100% with manual reading, and in 64–100% by immunodot assay. Receiver operating characteristic curve analysis of fluorescent intensity revealed a high sensitivity and specificity for automated reading of AAb with an agreement ranging from 90% to 95% between manual and automated interpretation (kappa 0.554–0.69) for systemic sclerosis and myositis, respectively. This study demonstrates a good correlation between manual and automated interpretation of AAb including ANA and ACyA in patients with autoimmune diseases. Full automation of HEp-2 cell assay reading may minimize errors in ANA pattern interpretation and thus help in the standardization of ANA assessment.

Keywords

Antinuclear antibodies Autoimmune diseases Automated ANA screening ELISA HEp-2 cells 

Abbreviations

AAb

Autoantibody

ANA

Antinuclear antibody

ACyA

Anticytoplasmatic antibody

IIF

Indirect immunofluorescence

ELISA

Enzyme-linked immunosorbent immunoassays

SLE

Systemic lupus erythematosus

DM

Dermatomyositis

PM

Polymyositis

SSc

Systemic sclerosis

Jo-1

Histidyl-tRNA synthetase

PL-7

Threonyl-tRNA synthetase

PL-12

Alanyl-tRNA synthetase

SRP

Signal recognition particle

ROC

Receiver operating characteristics

MSA

Mitotic spindle apparatus

AUC

Area under curve

Copyright information

© Clinical Rheumatology 2011

Authors and Affiliations

  • Shaye Kivity
    • 1
    • 2
    • 3
  • Boris Gilburd
    • 1
  • Nancy Agmon-Levin
    • 1
  • Marina Garcia Carrasco
    • 1
    • 7
  • Yaron Tzafrir
    • 1
  • Yael Sofer
    • 4
  • Matilda Mandel
    • 5
  • Thomas Buttner
    • 8
  • Dirk Roggenbuck
    • 8
  • Marco Matucci-Cerinic
    • 9
  • Katalin Danko
    • 10
  • Marcos López Hoyos
    • 11
  • Yehuda Shoenfeld
    • 1
    • 6
    • 12
  1. 1.The Zabludowicz Center for Autoimmune DiseasesTel-Aviv UniversityTel-AvivIsrael
  2. 2.Rheumatology UnitTel-Aviv UniversityTel-AvivIsrael
  3. 3.Department of Medicine ‘A & C’Tel-Aviv UniversityTel-AvivIsrael
  4. 4.Department of Medicine ‘A & C’Sheba Medical CenterTel HashomerIsrael
  5. 5.Blood CenterTel-Aviv UniversityTel-AvivIsrael
  6. 6.Department of Medicine B, Sheba Medical Center, Tel-Hashomer, Sackler Faculty of MedicineTel-Aviv UniversityTel-AvivIsrael
  7. 7.Department of RheumatologyClinical Hospital Buenos Aires UniversityBuenos AiresArgentina
  8. 8.GA Generic Assays GmbHBerlinGermany
  9. 9.Department of Biomedicine, Division of Rheumatology AOUCFlorenceItaly
  10. 10.Medical and Health Science CenterUniversity of DebrecenDebrecenHungary
  11. 11.Servicio Inmunología Hospital Universitario Marqués de aldecillaSantanderSpain
  12. 12.Incumbent of the Laura Schwarz-Kip Chair for Research of Autoimmune DiseasesTel-Aviv UniversityTel-AvivIsrael

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