Efficiency of different strategies to detect autoantibodies to extractable nuclear antigens.

Abstract Autoantibodies to extractable nuclear antigens (anti-ENA) are identified mainly in samples positive for antinuclear antibodies (ANA). Although the method of choice for ANA screening is indirect immunofluorescence (IIF), several techniques are available to detect anti-ENA. The aim of this study was to compare the efficiency of five different strategies to determine anti-ENA. During a 2-year period we screened ANA in 30375 samples with IIF, and the 4475 samples ANA positive were tested for anti-ENA by double immune diffusion screening or fluoroenzymeimmunoassay (Screening FI); anti-ENA specificities were then determined by line immunoassay (LIA) or fluoroenzymeimmunoassay (FI). We compared five strategies that involved FI or LIA identification of anti-ENA with or without prior screening, or an algorithm that combined fluorescence pattern, number of anti-ENA specificities requested by the clinician and ANA dilution titer. One cost unit (CU) was defined as the cost of 1 test of ANA determination. We detected 553 anti-ENA positive samples. The most efficient strategy was the algorithm, at a cost of 3.3 CU per sample processed, the second most efficient strategy was screening plus FI identification (cost = 3.8 CU), and the third most efficient strategy was screening plus LIA identification (cost = 3.9 CU). The fourth most efficient strategy was FI identification without prior screening (13.3 CU per sample) and the least efficient was LIA identification without prior screening (13.6 CU per sample). In conclusion, an algorithm that combined techniques for detection, ANA titer, fluorescence pattern and number of specificities requested was the most efficient strategy for determining anti-ENA.

[1]  H. Kunkel,et al.  Characteristics of a soluble nuclear antigen precipitating with sera of patients with systemic lupus erythematosus. , 1966, Journal of immunology.

[2]  R. Derksen,et al.  Cost-effective detection of non-antidouble-stranded DNA antinuclear antibody specificities in daily clinical practice. , 2006, Rheumatology.

[3]  Daniel H Solomon,et al.  Evidence-based guidelines for the use of immunologic tests: antinuclear antibody testing. , 2002, Arthritis and rheumatism.

[4]  B. Velkeniers,et al.  Diagnostic accuracy of the FIDIS multiplex fluorescent microsphere immunodetection system for anti-extractable nuclear antigen (ENA) antibodies in connective tissue diseases , 2007, Clinical chemistry and laboratory medicine.

[5]  A. H. Coons,et al.  Fluorescent Antibody Studies with Agents of Varicella and Herpes Zoster Propagated in vitro.∗ , 1954, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[6]  R. B. Dinkelaar,et al.  Different Strategies in the Laboratory Diagnosis of Autoimmune Disease: Immunofluorescence, Enzyme-Linked Immunosorbent Assay or Both? , 1999, Annals of clinical biochemistry.

[7]  Anne-Marie Rouquette,et al.  Evaluation of the new multiplexed immunoassay, FIDIS, for simultaneous quantitative determination of antinuclear antibodies and comparison with conventional methods. , 2003, American journal of clinical pathology.

[8]  M. Sticherling,et al.  Comparison of Different Test Systems for Simultaneous Autoantibody Detection in Connective Tissue Diseases , 2005, Annals of the New York Academy of Sciences.

[9]  M. Tampoia,et al.  Application of a combined protocol for rational request and utilization of antibody assays improves clinical diagnostic efficacy in autoimmune rheumatic disease. , 2007, Archives of pathology & laboratory medicine.

[10]  A. Wiik,et al.  Clinical optimization and multicenter validation of antigen-specific cut-off values on the INNO-LIAr ANA Update for the detection of autoantibodies in connective tissue disorders , 2002, Clinical and experimental rheumatology.

[11]  A. Ballabio,et al.  An Italian Multicenter Study for Application of a Diagnostic Algorithm in Autoantibody Testing , 2009, Annals of the New York Academy of Sciences.

[12]  P. Miescher,et al.  L'absorption du facteur «L.E.» par des noyaux cellulaires isolés , 1954, Experientia.

[13]  D Gianola,et al.  Multicenter validation of recombinant, natural and synthetic antigens used in a single multiparameter assay for the detection of specific anti-nuclear autoantibodies in connective tissue disorders. , 1999, Clinical and experimental rheumatology.

[14]  Marco Pradella,et al.  Guidelines for the laboratory use of autoantibody tests in the diagnosis and monitoring of autoimmune rheumatic diseases. , 2002, American journal of clinical pathology.

[15]  Marcos López-Hoyos,et al.  Performance of Antinuclear Antibody Connective Tissue Disease Screen , 2007, Annals of the New York Academy of Sciences.

[16]  R. Maini,et al.  Manual of Biological Markers of Disease , 1996, Springer Netherlands.

[17]  W. V. van Venrooij,et al.  The consensus workshops for the detection of autoantibodies to intracellular antigens in rheumatic diseases. , 1991, Journal of immunological methods.

[18]  E. Tan,et al.  Autoantibodies in the diagnosis of systemicrheumatic diseases , 1995 .

[19]  T. Kveder,et al.  Counterimmunoelectrophoresis and immunodiffusion for the detection of antibodies to soluble cellular antigens , 1996 .

[20]  P. Miescher,et al.  [Absorption of L. E. factor by isolated cell nuclei]. , 1954, Experientia.

[21]  M. Tampoia,et al.  Application of a diagnostic algorithm in autoantibody testing: assessment of clinical effectiveness and economic efficiency. , 2003, Clinica chimica acta; international journal of clinical chemistry.

[22]  T. Tomasi,et al.  Characterization of a soluble cytoplasmic antigen reactive with sera from patients with systemic lupus erythmatosus. , 1969, Journal of immunology.

[23]  S. Adelstein,et al.  High-Quality, Cost-Effective Strategy for Detection of Autoantibodies to Extractable Nuclear Antigens , 2001, Clinical Diagnostic Laboratory Immunology.

[24]  I. Peene,et al.  Detection of specific antinuclear reactivities in patients with negative anti-nuclear antibody immunofluorescence screening tests. , 2002, Clinical chemistry.

[25]  J. Koziol,et al.  Range of antinuclear antibodies in "healthy" individuals. , 1997, Arthritis and rheumatism.

[26]  N. Bizzaro,et al.  Variability between methods to determine ANA, anti-dsDNA and anti-ENA autoantibodies: a collaborative study with the biomedical industry. , 1998, Journal of immunological methods.