Abstract
The methods for detecting and measuring autoantibodies have evolved markedly in recent years, encompassing three generations of analytical technologies. Many different immunoassay methods have been developed and used for research and laboratory practice purposes, from the early conventional (or monoplex) analytical methods able to detect single autoantibodies to the more recent multiplex platforms that can quantify tens of molecules. Although it has been in use for over 50 years, indirect immunofluorescence remains the standard method for research on many types of autoantibodies, due to its characteristics of diagnostic sensitivity and also to recent technological innovations which permit it a greater level of automation and standardization. The recent multiplex immunometric methods, with varying levels of automation, present characteristics of higher diagnostic accuracy, but are not yet widely diffused in autoimmunology laboratories due to the limited number of autoantibodies that are detectable, and due to the high cost of reagents and systems. Technological advancement in autoimmunology continues to evolve rapidly, and in the coming years new proteomic techniques will be able to radically change the approach to diagnostics and possibly also clinical treatment of autoimmune diseases. The scope of this review is to update the state of the art of technologies and methods for the measurement of autoantibodies, with special reference to innovations in indirect immunofluorescence and in multiple proteomic methods.
Introduction
Circulating autoantibodies against a wide number of structural and functional molecules present in ubiquitous or tissue-specific cells are considered useful markers of autoimmune diseases (AIDs). The identification of these antigen-antibody systems with the contemporary development of laboratory methods for the detection and measurement of autoantibodies is considered one of the milestones in the history of clinical immunology over the last 60 years.
In this field, two categories of methods, based on microscopy and biochemistry principles – and therefore defined as morphologic and immunologic methods – have had great importance in the progress of laboratory medicine. These technological advances were transferred to the field of autoimmunology since all the variants of the immunological methods have found applications in the diagnosis of autoimmune diseases. A synthetic classification of these technologies is based on the principle of multiplicity of detection and distinguishes among conventional monoplex methods and multiplex methods according to the capacity to measure one antibody or several simultaneously [1]. Table 1 shows a (probably incomplete) list of these methods.
Classification of immunoassay methods for the detection and measurement of autoantibodies.
| Methods | Abbreviation |
|---|---|
| 1st generation monoplex | |
| Double immunodiffusion | ID |
| Complement fixation | CF |
| Indirect immunofluorescence | IIF |
| Passive agglutination | PHA, LPA |
| Radio-immunoprecipitation | RIPA |
| Western blot | WB |
| 2nd–3rd generation monoplex | |
| Radioimmunoassay-Immunoradiometric assay | RIA-IRMA |
| Radio-receptor assay | RRA |
| Immunoenzymatic assay-Immunoenzymometric assay | ELISA-IEMA |
| Immunoblot | IB |
| Immunodot | DB |
| Chemiluminescence immunoassay | CLIA-ILMA |
| Fluorimetric immunoassay | FIA-IFMA |
| Multiplex | |
| Addressable microbeads immunoassay | MBA |
The saga of immunodiagnostics originates in the late 1940s or early 1950s, with the discovery of demonstrable phenomena in vitro, made possible by the use of morphological methods. The first phenomenon connected with the presence of autoantibodies (LE phenomenon or LE test – reaction of opsonization of anti-nucleosome antibodies) was discovered by means of the common techniques of light microscopy [2]. Next, following the first applications of the method for identifying bacteria using conjugated antibodies with fluorescent tracers, for the first time antinuclear antibodies (ANA) were detected in sera using the indirect immunofluorescence technique (IIF) on antigenic microspots coated to slides [3] and later on cellular substrates made from human tissues and from isolated cells [4].
Thus, IIF was the first monoplex technique applied to the detection of autoantibodies. In some applications however, (i.e., for ANA) it is effectively a multiplex method, given that it is able to detect more than 60 autoantibodies simultaneously, by means of the identification of at least 26 different cellular patterns [5].
Next to IIF, also belonging to the first generation, are other qualitative immunochemical methods (immunodiffusion, complement fixation, passive agglutination, counterimmunoelectrophoresis, immunoprecipitation) developed during the year 1957 (the “golden year of immunodiagnostics”) [6–11]. During the 1970s and 1980s additional evolution took place in qualitative methods, with the introduction of immunoblot or dot-blot, and second-generation quantitative immunometric assays (IMA) were introduced [radioimmunoassay and its variants, immunoenzymatic assay and its variants, fluoroimmunologic assay, and immunochemiluminescent assay, reviewed in [1]].
Since the turn of the millennium, research has produced an additional rapid advance in diagnostic technology for detection and quantification of autoantibodies. The reasons for this technological revolution have been given as:
the increment of awareness of the physiopathogenetic and diagnostic role of autoantibodies in systemic and organ-specific autoimmune diseases (autoantibodies as diagnostic criteria);
the refinement of procedures for identifying and purifying the target autoantigens of the autoimmune reaction, with special reference to recombinant DNA technologies;
the application of quantitative determination of autoantibodies by automated analytic systems, initially introduced for determining hormones, tumor markers, metabolytes, antibodies, etc.;
the development of proteomic technologies, able to detect simultaneously a high number of autoantibodies in the same sample (multiplexing).
The occurrence of these processes in clinical laboratories has led to the advent of a new era in the measurement of autoantibodies, with exponential increase in analytic capacity and corresponding rise in the volume of test requests for the diagnosis of AIDs. This evolution has brought about the rapid disposal of 1st and 2nd generation immunoassay methods to the benefit of automated 3rd generation immunoassay methods [12] (Figure 1).
The evolution of technologies for autoantibody detection: the role of automation (see text for legends).
The aim of this review is to define the current state of technologies available in the autoimmunology laboratory (with special concern to the morphologic and proteomic technologies) with a view to updating our previous review [1].
Multiplex immunoassay platforms: a real improvement of the diagnostic power of autoantibody testing?
From the beginning of the third millennium, based on progress in the development of the genome, multiple measurement of genetic products has become a characteristic of laboratory methodology, defined with the suffix “omics”. Similar to other -omics, (e.g., ribonomics, metabolomics, etc.) proteomics – meaning the science which studies, on a large scale, the expression, the function and the interaction of protein – permits the parallel analysis of hundreds of different products, in this case antigens/antibodies, in minimal quantities of biological fluids. In this sense the discipline is held to represent the keystone for thoroughly investigating some acquired human diseases, in the fields of oncology, infectious diseases, immunology, and so on. Consequently, this has assisted the production of numerous analytic methods of both research and commercial application, for the simultaneous quantification of analytes of proteic nature (e.g., hormones, bio-humoral indicators, cytokines, antibodies, autoantibodies, etc.) by using new technologies, some of which can be considered as comprising a new category of immunometric methods [13].
Given the number and complexity of molecules involved in the activities of the immune system, the study of such molecules seemed an ideal field of application for proteomic techniques [14–16], the objective of which is to be able to study the entire autoimmune process rather than their single components. Among the many systems designed, some microarrays have found application for research on autoantibody profiling of AIDs and, in particular, planar and non-planar autoantigenic arrays (in suspension) [1, 17, 18].
Among the planar arrays we note systems made up of microspots on glass slides, on polystyrene microplates or nitrocellulose membranes and linear immunoblot systems on nitrocellulose membranes (Table 2). The first are based on techniques originally proposed by Ekins and Chu [19] and still hold a relevant importance for their simplicity and flexibility. There are not yet commercial applications available for these, but there are notable studies confirming their reliability [20–22].
Planar autoantigen arrays for autoantibody detection.
| Support | Format | Detection system | Autoantibodies tested (no. and type) | Estimated throughput (test/h) | Commercial product |
|---|---|---|---|---|---|
| Glass slide | Microspot | L | 115 | ~10.000 | – |
| Glass slide | Microspot | F | 196 | ~10.000 | – |
| Glass slide | Microspot | F | 650 | ~10.000 | – |
| Glass slide | Microspot | F | 225 | ~10.000 | – |
| Microplate | Microspot | C | 15 | ND | – |
| Nitrocellulose membrane | Microspot | C | 30 | ND | – |
| Nitrocellulose membrane | Line-blot | C | 15 (ENA) | ~50 | Inno-LIA |
| Nitrocellulose membranes | Line-blot | C | 13 (ENA) | ~50 | RecomLine |
| Nitrocellulose membrane | Line-blot | C | 11 (ENA) | ~50 | ANA-LIA |
| Nitrocellulose membrane | Nanodot | L | 10 (ENA) | ND | NALIA |
C, colorimetry; F, fluorescence; L, chemiluminescence; ND, not declared.
The second are based on the linear Western blot technique (line immunoblot), with electrophoretic migration on nitrocellulose membranes. The method has found numerous commercial applications and there is clinical validation of the systems conducted on large sample numbers, including multicentric studies [23–27]. In general, line immunoblot offers diagnostic and clinical accuracy comparable to, if not superior to, conventional immunometric methods. Nevertheless it is our opinion that such technologies are not to be considered true microarrays, given the macroscopic dimensions of the transferred proteins and the limits to the number and variety of antigens applicable on the planar solid phase of these systems.
Among the non-planar arrays (Table 3) there have been developed systems in suspension that use microparticles recognized by laser nephelometry [28] or laser fluorimetry in flow cytometry [29]. This last technology offers many commercial applications and has been validated widely by numerous clinical studies, conducted around the world. The principal commercial systems, based on fluorescent microbeads technology, are represented by the following: FIDIS (BioMedical Diagnostics, Marne la Vallee, France); AtheNA Multi-Lite (Zeus Diagnostics, Raritan, NJ, USA); QuantaPlex (Instrumentation Laboratory, Barcelona, Spain); and BioPlex 2200 (Bio-Rad Laboratories, Hercules, CA, USA) (Table 3).
Main commercial multiplex platforms and their market diffusion.
| Manufacturer | System | Profiles/autoantibodies detected (n) | Market diffusion |
|---|---|---|---|
| BMD, France | FIDIS | ANA (9), Rheuma (2), Vasculitis (2), Celiac (3), Thyroid (2) | + |
| IL (INOVA), Spain | Quanta Plex | ANA (8), Vasculitis (2), Celiac (3), Liver (5) | +/− |
| ZEUS, USA | Athena Multi-Lyte | ANA (10), Vasculitis (3), Celiac (2), Thyroid (2) | ++ |
| BIO-RAD, USA | Bioplex 2200 | ANA (13), ANCA (3), CCP (1), APS (2), Celiac (3) | +++ |
The FIDIS system, which has a reasonable level of automation and was the first to be introduced on the market, is able to determine diverse autoantibody profiles for connective tissue diseases, antiphospholipid syndrome, vasculitides, rheumatoid arthritis, celiac disease, autoimmune gastritis and autoimmune thyropathies. Many validation studies on its characteristics are published in the literature [30–37].
The AtheNA Multi-lyte system is able to do autoantibody profiling for rheumatic diseases, vasculitides, autoimmune thyroditis and celiac disease. The automation level is similar to the FIDIS method. The many validation studies published attest to its good analytic reliability [36–48].
The QuantaPlex system is able to determine various autoantibody profiles for rheumatic diseases, celiac disease, vasculitides and autoimmune liver diseases and demonstrates, like the first two, high analytic reliability [35, 49–52].
The BioPlex 2200 system is of recent introduction, totally automated and with high analytic productivity, which is able to execute an autoantibody profile for rheumatic diseases, antiphospholipid syndrome, vasculitides and, more recently, celiac disease. Numerous clinical validation studies have been conducted on extensive case populations [53–65].
Enthusiastically welcomed at the time of its appearance in the clinical laboratory a few years ago, multiplexing should have allowed for overcoming several limitations of the conventional methods, including analytic problems (e.g., reduction of the volume of samples and reagents and containment of costs), logistical/managerial problems (e.g., simultaneous measuring in the same session and in the same analyte reaction environment, measured in real time with several methods), and physiopathological problems (e.g., association of markers in pathology-oriented or organ-oriented profiling) [16–18] and should have found extensive application in the routine activities of the autoimmunology laboratory. In any case, at this time the biomedical industry seems oriented to repropose in multiplex versions the autoantibody profiles already consolidated for the principal AIDs, with the only intention being to achieve diagnostic usage limited to the most common autoantibody specificities. This is the principal motive for the slow and extended penetration of these systems into routine diagnostics, where they are substituting for conventional methods. However, a truly flexible autoantibody profile is not yet available to cover the 30–50 most common and important autoantibodies. Such a profile would be used in the laboratory simply for evaluating the autoimmune status and autoantibody profile of the patient, for predictive, monitoring, and therapeutic purposes [66].
The renaissance of IIF
Thanks to the use of various tissue and cellular substrates (e.g., human laryngeal HEp-2 carcinoma cells, Crithidia luciliae hemoflagellates, human neutrophil granulocytes, triple-tissue substrate composed of liver, kidney, and stomach of rat or mouse, monkey esophagus, human umbilical cord, human or animal pancreas, human or animal adrenal gland, pituitary, esophagus, bladder, monkey nerve tissue, etc.), IIF has permitted the detection over the years of an extensive series of specific autoantibodies directed against cellular autoantigens. This method, in time, became a consolidated and universally diffused procedure for detecting patients affected by AIDs, with differentiated use in the different AIDs, according to analytical sensitivity and specificity of the different types of substrates. Currently, the IIF method is considered the reference method for ANA and anti-neutrophil cytoplasmic antibodies (ANCA) screening and a confirmatory test for anti-DNA antibody detection [67–70]. However, the method is burdened by some unfavorable features: the need for expert morphologists, the subjectivity of interpretation, and the low degree of standardization and automation [71, 72]. Because of these limitations and the progressive increase of ANA test requests in autoimmunology laboratories, in the last 15 years technology innovation of analytical platforms has offered alternative solutions to the ANA-IIF test based on the manual or automated monoplex IMA, mainly of the ELISA type, with the use of solid phases made up by a mixture of nuclear-cytoplasmic antigens. Experience published in the literature has demonstrated that these manual [73–84] or automatic [85–87] systems do not provide the same analytic accuracy as IIF, in particular for the presence of false negative results (up to 35% of cases) in case of rare autoantibodies [79].
Therefore, it is maintained that the IMA monoplex methods do not represent a substitute for IIF, not even in unusual analytic conditions or when faced with a high volume of test requests [78].
The introduction of multiplex methods (multiplex immunoassays), automated in varying degrees, which are able to simultaneously measure several ANA-related antibodies, has produced – especially in the USA – the hypothesis that they can be used as a screening platform for ANA testing as an alternative to IIF. However, the sensitivity of the ANA screening test with multiplex immunoassay is not yet adequate and the presence of false negative results with respect to IIF is not dissimilar to the occurrence of the previously-discussed immunometric methods [35,42, 56, 62], varying from 0.2% to 40.5% according to the population studied (Table 4). One of our recent studies conducted with a latest-generation multiplex system (BioPlex 2200) showed a higher sensitivity and a parallel specificity of the ANA-IIF (at 1.80 dilution) compared to the ANA screen on BioPlex 2200. In the study of 181 patients affected by autoimmune rheumatic diseases, the rate of false negatives has stabilized around 11.6% for the manual IIF method, vs. 14.9% for the BioPlex method (Table 5). In most cases, the patients had nucleolar or rare autoantibodies. In a group of 208 healthy and infectious disease patients, the specificity was similar between the two methods (Table 5).
Diagnostic sensitivity of indirect immunofluorescence (IIF) and multiplex immunoassay (MIA) methods for ANA testing.
| Author, year | Patients, n | IIF positive, % | MIA positive, % | Multiplex system |
|---|---|---|---|---|
| Nifli et al., 2006 [41] | 910 | 17.3 | 17.1 | AtheNA Multi-lyte |
| Bonilla et al., 2007 [42] | 53 | 90.6 | 49.1 | AtheNA Multi-lyte |
| Salamunic et al., 2008 [46] | 897 | 27.9 | 23.6 | AtheNA Multi-lyte |
| Hanly et al., 2010 [52] | 192 | 81.3 | 75.5 | BioPlex 2200 |
| Op de Beeck et al., 2012 [65] | 236 | 78.3 | 74.5 | BioPlex 2200 |
Sensitivity of methods for ANA detection in AIDs: comparison between IIF and BioPlex 2200.
| Patients | n | IIF positive (1:80) | Bioplex positive | ||
|---|---|---|---|---|---|
| n | % | n | % | ||
| SLE | 95 | 85 | 89.5 | 77 | 81.1 |
| SSc | 55 | 53 | 96.4 | 52 | 94.5 |
| SS | 18 | 18 | 100 | 18 | 100 |
| DM/PM | 8 | 4 | 50 | 7 | 87.5 |
| Vasculitis | 5 | 0 | 0 | 0 | 0 |
| Total | 181 | 160 | 88.4 | 154 | 85.1 |
| Blood donors | 120 | 7 | 5.8 | 6 | 5.0 |
| Infectious disease | 98 | 7 | 7.1 | 6 | 6.1 |
| Total | 208 | 14 | 6.7 | 12 | 5.8 |
SLE, systemic lupus erythematosus; SSc, systemic sclerosis; SS, Sjögren’s syndrome; DM/PM, dermatomyositis/polymyositis.
Following the recent statement that the IIF technique should be considered as the standard screening method for the detection of ANA [69], the biomedical industry has proposed technological solutions which significantly improve the automation for the procedure, not only in the preparation of substrates and slides but also in reading under the microscope. This innovation is based on the principles of digitalization of fluoroscopic images and on the classification of patterns using standardized approaches (automatic positive-negative pattern interpretations) using expert systems. These systems are based on the use of automated microscopes, robotized slide trays, high-sensitivity CCD video cameras, and software dedicated to acquisition and analysis of digital images [71, 88–92].
Currently, the advanced stages of experimentation are taking place on five commercial systems (Aklides, Medipan, Berlin, Germany; Nova View, Instrumentation Laboratory, Barcelona, Spain; Zenit G Sight, A. Menarini Diagnostics, Florence, Italy; Euroscope-Europattern, Euroimmun, Luebeck, Germany; Helios, Aesku.Diagnostics, Wendelsheim, Germany). Their characteristics are shown in Table 6.
Patterns detected by the screening/recognition IIF automated systems.
| System | Screening –/+ | Patterns, n | Type |
|---|---|---|---|
| Aklides | Yes | 5 | Homogeneous, speckled, nucleolar, centromeric, nuclear dots |
| Europattern | Yes | 6 | Homogeneous, speckled, nucleolar, centromeric, nuclear dots, cytoplasmic |
| G-Sight | Yes | 5 | Homogeneous, speckled, nucleolar, centromeric, nuclear dots, mytochondrial |
| Nova-View | Yes | 5 | Homogeneous, speckled, nucleolar, centromeric, nuclear dots |
| Helios | Yes | None | – |
We evaluated the diagnostic performance of some of these fully automated IIF interpretation systems in routine diagnostics, in comparison with the visual expert interpretation of IIF. A total number of 298 serum samples from patients with suspected autoimmune disease were collected from March to April 2011 for ANA testing in two different experiments, using Aklides (Medipan, Germany) and Europattern (Euroimmun, Germany) systems. Results demonstrated a very high agreement of the two automated systems with the manual IIF method (Table 7).
Diagnostic sensitivity of manual and automated IIF methods for ANA testing.
| System | Patients | Manual IIF no. positive (1:80) | Automated IIF no. positive | Agreement, % |
|---|---|---|---|---|
| Europattern | 116 | 70 | 70 | 100 |
| Aklides | 182 | 182 | 180 | 98.9 |
Our findings are consistent with recent literature data that have shown a high sensitivity (close to 100%) and a good agreement between the Aklides and Europattern systems and visual IIF interpretation for ANA, in patients with AIDs [93, 94].
At present, these test systems might be helpful as a screening tool, although the performance of reading of slides and automated systems needs to be further improved. The availability of these systems suggests that automation of cell-based IIF testing may improve standardization of ANA assessment and help to reduce variability among autoimmunology laboratories in the near future.
However, there is still insufficient evidence that the new automated platforms present the same careful analysis of the IIF patterns as the conventional IIF procedure: this is a critical point, because the recognition of immunofluorescence patterns not associated with systemic autoimmune diseases is helpful in the interpretation of a positive ANA result in healthy individuals [95]. Further studies are needed to define the specificity of automated IIF.
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors declare that this study was not financially supported by any pharmaceutical organization or industry and that that there are no conflicts of interest regarding the publication of this article.
Research funding: None declared.
Employment or leadership: None declared.
About the authors
Renato Tozzoli was born in Venice, Italy, in 1951. He is Head of the Department of Laboratory Medicine and Medical Director at the Clinical Pathology Laboratory, St. Mary of Angels Hospital, Pordenone, Italy. He graduated in Medicine and Surgery at the University of Bologna, and specialized in Endocrinology in 1984 at the Modena University and in Clinical Pathology in 1988 at Padua University. He has published over 250 papers in national (Italian) and international journals, including several convention reports, books, and book chapters. He is General Chairman of the Italian Society of Laboratory Medicine Study Groups; Member of the Italian Association for Autoimmune Research; Founder and President of the Italian Interdisciplinary Forum for Research in Autoimmune Diseases (FIRMA) and Deputy Editor of Autoimmunity Highlights (Springer).
Chiara Bonaguri was born in Forlì, Italy in 1957. She obtained a PhD degree in Biology in 1981 at the University of Bologna; in 1991 she specialized in Biochemistry and Clinical Chemistry at the University of Parma and in Clinical Immunopathology (2008) at the University of Milan. Since 1992, she has worked at the Laboratory of Clinical Immunology of the University Hospital of Parma; and in 2009 headed the Autoimmune Serology Section. She has taught Allergology and Clinical Immunology since 1999 at the School of Medicine of the University of Parma. She is the author of several publications in peer-reviewed journals on autoimmune diseases and is a member of the Study Group on Autoimmune Diseases of the Italian Society of Laboratory Medicine.
Alessandra Melegari was born in Parma, Italy in 1961. She obtained her BSc in Biological Sciences at the Universita’ delgi Studi di Parma in 1985 and her PhD in Food Chemistry and Technology in 1991. She was a biologist in various hospitals from 1992 to 1999 and a senior Diagnostic Biologist at the University Department of the Policlinico Hospital at Modena from 1999 to 2010. Since 2010 she has been a Senior Biologist in the Diagnostic Department of the NOCSAE at Modena Hospital and Head of Diagnostic Autoimmune Disease Activity. Alessandra is a teacher at the Post-graduate School of Rheumatology and Laboratory Medicine at the University of Modena where she instructs post-graduate students. She is active in many national and international congresses in laboratory medicine, and is the author and co-author of many articles on immunology, rheumatology and autoimmunity. Alessandra organized congresses on immunology in Modena in 2011 and 2012, and she has also organized courses for general physicians on autoimmune diseases.
Antonio Antico, was born in 1960. In 1987 he graduated in Medicine and later specialized in Clinical Pathology at the University of Padua. Currently he works as a medical doctor at the Laboratory of Clinical Pathology, General Hospital of Cittadella, Italy. Since 1993 he has mostly worked in the field of autoimmune/rheumatic diseases. He is a member of the Study Group on Autoimmunology of the Italian Society of Laboratory Medicine. He has published many scientific papers in international journals on clinical-serological aspects of connective tissue diseases, autoimmune liver diseases, autoimmune gastritis, celiac disease, autoimmune blistering skin diseases and autoimmune neurological diseases.
Danila Bassetti graduated in Medicine and Surgery in 1978 at the Bologna University (Italy), where she specialized in Hygiene and Preventive Medicine (Public Health and Laboratory) and Health Law. Since 1981 has directed the Serology section of the Public Health Laboratory in Trento (Italy) and since 1999 the Serology and Autoimmunity section at the Microbiology and Virology Unit of the S. Chiara Hospital in Trento. She is author of scientific articles on autoimmunity, infectivology and is Professor of Immunopathology at the Biomedical Laboratory Technician School of Verona University.
Nicola Bizzaro was born in Venice, Italy, in 1951. He is Head and Medical Director at the Laboratory of Clinical Pathology, San Antonio Hospital, Tolmezzo, Italy. He graduated in Medicine and Surgery at the University of Padua, and specialized in Clinical and Laboratory Hematology in 1987 and in Clinical Pathology in 1993 at Padua University. He has published over 440 papers in national (Italian) and international journals, including several convention reports, books, and book chapters. He is Chairman of the Italian Society of Laboratory Study Group on Autoimmune Diseases; Member of the Italian Association for Autoimmune Research; Founder of the Interdisciplinary Forum for Research in Autoimmune Diseases; Member of the Advisory Board of the International Congress on Autoimmunity; Chief Editor of the Italian Journal of Laboratory Medicine, the official Journal of the Italian Society of Laboratory Medicine; and Editor-in-Chief of Autoimmunity Highlights.
References
1. Tozzoli R. Recent advances in diagnostic technologies and their impact in autoimmune diseases. Autoimmun Rev 2007;6:334–40.10.1016/j.autrev.2007.01.005Search in Google Scholar
2. Hargraves MM, Richmond H, Morton R. Presentation of two bone marrow elements: the tart cell and the L.E. cell. Mayo Clin Proc 1948;23:25–8.Search in Google Scholar
3. Friou GJ. Clinical application of lupus serum-nucleoprotein reaction using fluorescent antibody technique. J Clin Invest 1957;86:890–6.Search in Google Scholar
4. Holborow EJ, Weir DM, Johnson GD. A serum factor in lupus erythematosus with affinity for tissue nuclei. Br Med J 1957;2:732–4.10.1136/bmj.2.5047.732Search in Google Scholar
5. Wiik A, Hoier-Madsen M, Forslid J, Charles P, Meyrowitsch J. Antinuclear antibodies: a contemporary nomenclature using HEp-2 cells. J Autoimmun 2010;35:276–90.10.1016/j.jaut.2010.06.019Search in Google Scholar
6. Robbins WC, Holman HR, Deicher H, Kunkel HG. Complement fixation with all nuclei and DNA in lupus erythematosus. Proc Soc Exp Biol Med 1957;96:575–9.10.3181/00379727-96-23545Search in Google Scholar
7. Trotter WR, Belyavin G, Waddams A. Precipitating and complement-fixing antibodies in Hashimoto’s disease. Proc Roy Soc Med 1957;50:961–2.Search in Google Scholar
8. Anderson JD, Goudie RB, Gray K. Autoantibodies in Addison’s disease. Lancet 1957;1:1123–4.10.1016/S0140-6736(57)91687-2Search in Google Scholar
9. Witebsky E, Rose NR, Teplan K. Chronic thyroiditis and immunization. J Am Med Assoc 1957;64:1439–43.10.1001/jama.1957.02980130015004Search in Google Scholar PubMed
10. Ceppellini R, Polli E, Celada F. A DNA-reacting factor in serum of a patient with lupus erythematosus diffusus. Proc Soc Exp Biol Med 1957;96:572–4.10.3181/00379727-96-23544Search in Google Scholar PubMed
11. Doniach D, Roitt IM. Auto-immunity in Hashimoto’s disease and its implications. J Clin Endocrinol Metab 1957;17:1293–304.10.1210/jcem-17-11-1293Search in Google Scholar PubMed
12. Tozzoli R, Bizzaro N. The clinical autoimmunologist and the laboratory autoimmunologist: the two sides of the coin. Autoimmun Rev 2012, doi:10–1016/J autrev.2012.02.011.10.1016/j.autrev.2012.02.011Search in Google Scholar PubMed
13. Ellington AA, Kullo IJ, Bailey KR, Klee GG. Antibody-based protein multiplex platforms. Technical and operational challenges. Clin Chem 2010;56:186–93.10.1373/clinchem.2009.127514Search in Google Scholar PubMed PubMed Central
14. Robinson WH, Steinman L, Utz PJ. Proteomic technologies for the study of autoimmune disease. Arthritis Rheum 2002;46:885–93.10.1002/art.10129Search in Google Scholar PubMed
15. Wu T, Mohan C. Proteomic toolbox for autoimmunity research. Autoimmun Rev 2009;8:595–8.10.1016/j.autrev.2009.01.019Search in Google Scholar PubMed
16. Fritzler MJ. Advances and applications of multiplexed diagnostic technologies in autoimmune diseases. Lupus 2006;15:422–7.10.1191/0961203306lu2327oaSearch in Google Scholar PubMed
17. Tozzoli R, Bizzaro N. Novel diagnostic methods for autoantibody detection. In: Shoenfeld Y, Gershwin ME, Meroni PL, editors. Autoantibodies. Amsterdam: Elsevier, 2006:77–82.Search in Google Scholar
18. Bizzaro N, Tozzoli R, Shoenfeld Y. Are we at a stage to predict autoimmune rheumatic diseases? Arthritis Rheum 2007;56:1736–44.10.1002/art.22708Search in Google Scholar PubMed
19. Ekins R, Chu F. Multianalyte microspot immunoassay: the microanalytical compact disk of the future. Ann Biol Clin (Paris) 1992;50:337–53.Search in Google Scholar
20. Robinson WH, DiGennaro C, Hueber W, Haab BB, Kamachi M, Dean EJ, et al. Autoantigen microarrays for multiplex characterization of autoantibody responses. Nat Med 2002;8: 295–301.10.1038/nm0302-295Search in Google Scholar PubMed
21. Hueber W, Kidd BA, Tomooka BH, Lee BJ, Bruce B, Fries JF, et al. Antigen microarray profiling of autoantibodies in rheumatoid arthritis. Arthritis Rheum 2005;52:2645–55.10.1002/art.21269Search in Google Scholar PubMed
22. McBride JD, Gabriel FG, Fordham J, Kolind T, Barcenas-Morales G, Isenberg DA, et al. Screening autoantibody profiles in systemic rheumatic disease with a diagnostic protein microarray that use a filtration assisted nanodot array luminometric immunoassay (NALIA). Clin Chem 2008;54:883–90.10.1373/clinchem.2007.098418Search in Google Scholar PubMed
23. Meheus L, van Venrooij WJ, Wiik A, Charles PJ, Tzioufas AG, Meyer O, 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. Clin Exp Rheumatol 1999;17: 205–14.Search in Google Scholar
24. Lopez-Longo FJ, Rodriguez-Mahou M, Escalona-Monge M, González CM, Monteagudo I, Carreño-Pérez L. Simultaneous identification of various antinuclear antibodies using an automated multiparameter line immunoassay system. Lupus 2003;12:623–9.10.1191/0961203303lu439oaSearch in Google Scholar PubMed
25. Pottel H, Wiik A, Locht H, Gordon T, Roberts-Thomson P, Abraham D, et al. Clinical optimization and multicenter validation of antigen-specific cut-off values on the INNO-LIA ANA update for the detection of autoantibodies in connective tissue disorders. Clin Exp Rheumatol 2004;22:579–88.Search in Google Scholar
26. Eissfeller P, Sticherling M, Scholz D, Hennig K, Luttich T, Motz M, et al. Comparison of different test systems for simultaneous antoantibody detection in connective tissue diseases. Ann N Y Acad Sci 2005;1050:327–39.10.1196/annals.1313.035Search in Google Scholar PubMed
27. Almeida Gonzalez D, Cabrera de Leon A, Rodriguez Perez MdelC, Brito Díaz B, González Hernández A, García García D, et al. Efficiency of different strategies to detect autoantibodies to extractable nuclear antigens. J Immunol Methods 2010;360:89–95.10.1016/j.jim.2010.06.013Search in Google Scholar PubMed
28. Bizzaro N, Bonelli F, Tonutti E, Villalta D, Tozzoli R. New coupled-particle light-scattering assay for detection of Ro/SSA (52 and 60 kilodaltons) and La/SSB autoantibodies in connective tissue diseases. Clin Diagn Lab Immunol 2001;8:922–5.10.1128/CDLI.8.5.922-925.2001Search in Google Scholar PubMed PubMed Central
29. Fulton RJ, McDade RL, Smith PL, Kienker LJ, Kettman JR Jr. Advanced multiplexed analysis with the FluoMetrix system. Clin Chem 1997;43:1749–56.10.1093/clinchem/43.9.1749Search in Google Scholar
30. Rouquette A-M, Desgruelles C, Laroche P. Evaluation of the new multiplexed immunoassay, FIDIS, for simultaneous quantitative determination of antinuclear antibodies and comparison with conventional methods. Am J Clin Pathol 2003;120:676–81.10.1309/GJHK0D24YDDXW0NFSearch in Google Scholar
31. Yiannaki E, Zintzaras E, Analatos A, Thedoridou C, Dalekos GN, Germenis AE. Evaluation of a microsphere-based flow cytometric assay for diagnosis of celiac disease. J Immunoassay Immunochem 2004;25:345–7.10.1081/IAS-200033832Search in Google Scholar PubMed
32. Abreu I, Laroche P, Bastos A, Issert V, Cruz M, Nero P, et al. Multiplexed immunoassay for detection of rheumatoid factors by FIDISTM technology. Ann N Y Acad Sci 2005;1050:357–63.10.1196/annals.1313.038Search in Google Scholar PubMed
33. Tozzoli R, Villalta D, Kodermaz G, Bagnasco M, Tonutti E, Bizzaro N. Autoantibody profiling of patients with autoimmune thyroid disease using a new multiplexed immunoassay method. Clin Chem Lab Med 2006;44:837–42.10.1515/CCLM.2006.137Search in Google Scholar PubMed
34. Gonzalez C, Garcia-Berrocal B, Talavan T, Casas ML, Navajo JA, Gonzalez-Buitrago JM. Clinical evaluation of a microsphere bead-based flow cytometry assay for the simultaneous determination of anti-thyroid peroxidase and anti-thyroglobulin antibodies. Clin Biochem 2005;38:966–72.10.1016/j.clinbiochem.2005.08.004Search in Google Scholar PubMed
35. Damoiseaux J, Vaessen M, Knapen Y, Csernok E, Stegeman CA, Van Paassen P, et al. Evaluation of the FIDIS vasculitis multiplex immunoassay for diagnosis and follow-up of ANCA-associated vasculitis and Goodpasture’s disease. Ann N Y Acad Sci 2007;1109:454–63.10.1196/annals.1398.051Search in Google Scholar PubMed
36. Copple SS, Martins TB, Masterson C, Joly E, Hill HR. Comparison of three multiplex immunoassays for detection of antibodies to extractable nuclear antibodies using clinically defined sera. Ann N Y Acad Sci 2007;1109:464–72.10.1196/annals.1398.052Search in Google Scholar PubMed
37. Albon S, Bunn C, Swana G, Karim Y. Performance of a multiplex assay compared to enzyme and precipitation methods for anti-ENA testing in systemic lupus and systemic sclerosis. J Immunol Methods 2011;365:126–31.10.1016/j.jim.2010.12.010Search in Google Scholar PubMed
38. Gilburd B, Abu-Shakra M, Shoenfeld Y, Giordano A, Bocci EB, delle Monache F, et al. Autoantibodies profile in the sera of patients with Sjögren syndrome: the ANA evaluation – a homogeneous, multiplexed system. Clin Dev Immunol 2004;11:53–6.10.1080/10446670410001670490Search in Google Scholar PubMed PubMed Central
39. Shovman O, Gilburd B, Zandman-Goddard G, Yehiely A, Langevitz P, Shoenfeld Y. Multiplexed AtheNA multi-lyte immunoassay for ANA screening in autoimmune diseases. Autoimmunity 2005;38:105–9.10.1080/08916930400022707Search in Google Scholar PubMed
40. Zandman-Goddard G, Gilburd B, Shovman O, Blank M, Berdichevski S, Langevitz P, et al. The homogeneous multiplexed system – a new method for autoantibody profile in systemic lupus erythemathosus. Clin Dev Immunol 2005;12:107–11.10.1080/17402520500116723Search in Google Scholar PubMed PubMed Central
41. Nifli A-P, Notas G, Mamoulaki M, Niniraki M, Ampartzaki V, Theodoropoulos PA, et al. Comparison of a multiplex, bead-based fluorescent assay and immunofluorescence methods for the detection of ANA and ANCA autoantibodies in human serum. J Immunol Methods 2006;311:189–97.10.1016/j.jim.2006.02.004Search in Google Scholar PubMed
42. Bonilla E, Francis L, Allam F, Ogrinc M, Neupane H, Phillips PE, et al. Immunofluorescence microscopy is superior to fluorescent beads for detection of antinuclear antibody reactivity in systemic lupus erythematosus patients. Clin Immunol 2007;124:18–21.10.1016/j.clim.2007.04.010Search in Google Scholar PubMed PubMed Central
43. Caramaschi P, Ruzzenente O, Pieropan S, Volpe A, Carletto A, Bambara LM, et al. Determination of ANA specificity using multiplexed fluorescent microsphere immunoassay in patients with ANA positivity at high titres after infliximab treatment: preliminary results. Rheumatol Int 2007;27:649–54.10.1007/s00296-006-0271-8Search in Google Scholar PubMed
44. Avaniss-Aghajani E, Berzon S, Sarkissian A. Clinical value of multiplexed bead-based immunoassays for detection of autoantibodies to nuclear antigens. Clin Vaccine Immunol 2007;14:505–9.10.1128/CVI.00034-07Search in Google Scholar PubMed PubMed Central
45. Biagini RE, Parks CG, Smith JP, Sammons DL, Robertson SA. Analytical performance of the AtheNA Multilite ANA II assay in sera from lupus patients with multiple positive ANAs. Anal Bioanal Chem 2007;388:613–8.10.1007/s00216-007-1243-xSearch in Google Scholar PubMed
46. Salamunic I, Paukovic-Sekulic B, Galetovic A. Comparative analysis of multiplex AtheNA multi-lyte ANA test system and conventional laboratory methods to detect autoantibodies. Biochemia Medica 2008;18:88–98.10.11613/BM.2008.010Search in Google Scholar
47. Eriksson C, Kokkonen H, Johansson M, Hallmans G, Wadell G, Rantapää-Dahlqvist S. Autoantibodies predate the onset of systemic lupus erythematosus in northern Sweden. Arthritis Res Ther 2011:13:R30.10.1186/ar3258Search in Google Scholar PubMed PubMed Central
48. Giovanella L, Toffalori E, Tozzoli R, Caputo M, Ceriani L, Verburg FA. Multiplexed immunoassay of thyroglobulin autoantibodies in patients with differentiated thyroid carcinoma. Head Neck 2011. doi:10.1002/hed 21933.10.1002/hed21933Search in Google Scholar
49. Martins TB, Burlingame R, von Muhlen C, Jaskowski TD, Litwin CM, Hill HR. Evaluation of multiplexed fluorescent microsphere immunoassay for detection of autoantibodies to nuclear antigens. Clin Diagn Lab Immunol 2004;11:1054–9.10.1128/CDLI.11.6.1054-1059.2004Search in Google Scholar PubMed PubMed Central
50. Mahler M, Stinton LM, Fritzler MJ. Improved serological differentiation between systemic lupus erythematosus and mixed connective tissue disease by use of an SmD3 peptide-based immunoassay. Clin Diagn Lab Immunol 2005;12:107–13.10.1128/CDLI.12.1.107-113.2005Search in Google Scholar PubMed PubMed Central
51. Fritzler MJ, Behmanesh F, Fritzler ML. Analysis of human sera that are polyreactive in an addressable laser bead immunoassay. Clin Immunol 2006;120:349–56.10.1016/j.clim.2006.03.007Search in Google Scholar PubMed
52. Hanly JG, Su L, Farewell V, Fritzler MJ. Comparison between multiplex assays for autoantoantibody detection in systemic lupus erythematosus. J Immunol Methods 2010;358:75–80.10.1016/j.jim.2010.04.005Search in Google Scholar PubMed
53. Shovman O, Gilburd B, Barzilai O, Shinar E, Larida D, Zandman-Goddard G, et al. Evaluation of the BioPlex 2200 ANA screen: analysis of 510 healthy subjects: incidence of natural/predictive autoantibodies. Ann N Y Acad Sci 2005;1050:380–8.10.1196/annals.1313.120Search in Google Scholar PubMed
54. Binder SR, Genovese MC, Merrill JT, Morris RI, Metzger AL. Computer-assisted pattern recognition of autoantibody results. Clin Diagn Lab Immunol 2005;12:1353–7.10.1128/CDLI.12.12.1353-1357.2005Search in Google Scholar PubMed PubMed Central
55. Moder KG, Wener MH, Weisman MH, Ishimori ML, Wallace DJ, Buckeridge DL, et al. Measurement of antinuclear antibodies by multiplex immunoassay: a prospective, multicenter clinical evaluation. J Rheumatol 2007;34:978–86.Search in Google Scholar
56. Desplat-Jego S, Bardin N, Larida B, Sanmarco M. Evaluation of the BioPlex 2200 ANA screen for the detection of antinuclear antibodies and comparison with conventional methods. Ann N Y Acad Sci 2007;1109:245–55.10.1196/annals.1398.030Search in Google Scholar PubMed
57. Tozzoli R, Barzilai O, Ram M, Villalta D, Bizzaro N, Sherer Y, et al. Infections and autoimmune thyroid diseases: parallel detection of antibodies against pathogens with proteomic technology. Autoimmun Rev 2008;8:112–5.10.1016/j.autrev.2008.07.013Search in Google Scholar PubMed
58. Kaul R, Johnson K, Scholz H, Marr G. Performance of the BioPlex 2200 autoimmune vasculitis kit. Autoimmun Rev 2009;8:224–7.10.1016/j.autrev.2008.07.033Search in Google Scholar PubMed
59. Bardin N, Desplat-Jego S, Daniel L, Jourde Chiche N, Sanmarco M. BioPlex 2200 multiplexed system: simultaneous detection of anti-dsDNA and anti-chromatin antibodies in patients with systemic lupus erythematosus. Autoimmunity 2009;42:63–8.10.1080/08916930802354906Search in Google Scholar PubMed
60. Orbach H, Amitai N, Barzilai O, Boaz M, Ram M, Zandman-Goddard G, et al. Autoantibody screen in inflammatory myopathies: high prevalence of antibodies to gliadin. Ann N Y Acad Sci 2009;1173:174–9.10.1111/j.1749-6632.2009.04810.xSearch in Google Scholar PubMed
61. Lidar M, Langevitz P, Barzilai O, Ram M, Porat-Katz BS, Bizzaro N, et al. Infectious serologies and autoantibodies in inflammatory bowel disease: insinuations at a true pathogenic role. Ann N Y Acad Sci 2009;1173:640–8.10.1111/j.1749-6632.2009.04673.xSearch in Google Scholar PubMed
62. Hanly JG, Thompson K, McCurdy G, Fougere L, Theriault C, Wilton K. Measurement of autoantibodies using multiplex methodology in patients with systemic lupus erythematosus. J Immunol Methods 2010;352:147–52.10.1016/j.jim.2009.10.003Search in Google Scholar PubMed
63. Shapira Y, Poratkatz BS, Gilburd B, Barzilai O, Ram M, Blank M, et al. Geographical differences in autoantibodies and anti-infectious agents antibodies among healthy adults. Clin Rev Allergy Immunol 2012;42:154–63.10.1007/s12016-010-8241-zSearch in Google Scholar PubMed
64. Kim Y, Park Y, Lee EY, Kim HS. Comparison of automated multiplexed bead-based ANA screening assay with ELISA for detecting five common anti-extractable nuclear antigens and anti-dsDNA in systemic rheumatic diseases. Clin Chim Acta 2012;413:308–11.10.1016/j.cca.2011.10.017Search in Google Scholar PubMed
65. Op De Beeck K, Vermeesch P, Verschueren P, Westhovens R, Marien G, Blockmans D, et al. Antinuclear antibody detection by automated multiplex immunoassay in untreated patients at the time of diagnosis. Autoimmun Rev 2012 (Epub ahead of print).10.1016/j.autrev.2012.02.013Search in Google Scholar PubMed
66. Tozzoli R. The diagnostic role of autoantibodies in the prediction of organ specific autoimmune diseases. Clin Chem Lab Med 2008;46:577–87.10.1515/CCLM.2008.138Search in Google Scholar
67. Solomon DH, Kavanaugh AJ, Schur PH. Evidence-based guidelines for the use of immunologic tests: antinuclear antibody testing. Arthritis Rheum 2002;47:434–44.10.1002/art.10561Search in Google Scholar PubMed
68. Tozzoli R, Bizzaro N, Tonutti E, Villalta D, Bassetti D, Manoni F, et al. Guidelines for the laboratory use of autoantibody tests in the diagnosis and monitoring of autoimmune rheumatic diseases. Am J Clin Pathol 2002;117:316–24.10.1309/Y5VF-C3DM-L8XV-U053Search in Google Scholar PubMed
69. Meroni PL, Schur PH. ANA screening: an old test with new recommendations. Ann Rheum Dis 2010;69:1420–2.10.1136/ard.2009.127100Search in Google Scholar PubMed
70. Savige JF, Gillis DF, Benson E, Davies D, Esnault V, Falk RJ, et al. International Consensus Statement on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA). Am J Clin Pathol 1999;111:507–13.10.1093/ajcp/111.4.507Search in Google Scholar PubMed
71. Rigon A, Soda P, Zennaro D, Iannello G, Afeltra A. Indirect immunofluorescence in autoimmune diseases: assessment of digital images for diagnostic purpose. Cytometry B 2007;72B:472–7.10.1002/cyto.b.20356Search in Google Scholar PubMed
72. Fritzler MJ. The antinuclear antibody test: last or lasting gasp? Arthritis Rheum 2011;63:19–22.10.1002/art.30078Search in Google Scholar PubMed
73. Jaskowski TD, Schroder C, Martins TB, Mouritsen CL, Litwin CM, Hill HR. Screening of antinuclear antibodies by enzyme immunoassay. Am J Clin Pathol 1996;105:468–73.10.1093/ajcp/105.4.468Search in Google Scholar PubMed
74. Emlen W, O’Neill L. Clinical significance of antinuclear antibodies. Arthritis Rheum 1997;40:1612–8.10.1002/art.1780400910Search in Google Scholar PubMed
75. Gniewek RA, Stites DP, McHugh TM, Hilton JF, Nakagawa M. Comparison of antinuclear antibody testing: immunofluorescence assay versus enzyme immunoassay. Clin Diagn Lab Immunol 1997;4:185–8.10.1128/cdli.4.2.185-188.1997Search in Google Scholar PubMed PubMed Central
76. Homburger HA, Cahen YD, Griffiths J, Jacob GL. Detection of antinuclear antibodies: comparative evaluation of enzyme immunoassay and indirect immunofluorescence methods. Arch Pathol Lab Med 1998;122:993–9.Search in Google Scholar
77. Rondeel JM, van Gelder W, van der Leeden H, Dinkelaar RB. Different strategies in the laboratory diagnosis of autoimmune disease: immunofluorescence, enzyme-linked immunosorbent assay or both? Ann Clin Biochem 1999;36:189–95.10.1177/000456329903600209Search in Google Scholar PubMed
78. Olaussen E, Rekvig OP. Screening tests for antinuclear antibodies: selective use of central nuclear antigens as a rational basis for screening by ELISA. J Autoimmun 1999;13: 95–102.10.1006/jaut.1999.0295Search in Google Scholar PubMed
79. Ulvestad E. Performance characteristics and clinical utility of a hybrid ELISA for detection of ANA. APMIS 2001;109:217–22.10.1034/j.1600-0463.2001.090305.xSearch in Google Scholar PubMed
80. Bernardini S, Infantino M, Bellicampi L, Nuccetelli M, Afeltra A, Lori R, et al. Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay. Clin Chem Lab Med 2004;42:1155–60.10.1515/CCLM.2004.235Search in Google Scholar PubMed
81. Tonutti E, Bassetti D, Piazza A, Visentini D, Poletto M, Bassetto F, et al. Diagnostic accuracy of ELISA methods as an alternative screening test to indirect immunofluorescence for the detection of antinuclear antibodies. Evaluation of five commercial kits. Autoimmunity 2004;37:171–6.10.1080/08916930310001657010Search in Google Scholar PubMed
82. Fenger M, Wiik A, Hoier-Madsen M, Lykkegaard JJ, Rozenfeld T, Hansen MS, et al. Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis. Clin Chem 2004;50:2141–7.10.1373/clinchem.2004.038422Search in Google Scholar PubMed
83. Sinclair D, Saas M, Williams D, Hart M, Goswami R. Can an ELISA replace immunofluorescence for the detection of anti-nuclear antibodies? The routine use of anti-nuclear antibody screening ELISAs. Clin Lab 2007;53:183–91.Search in Google Scholar
84. Lopez-Hoyos M, Rodriguez-Valverde V, Martinez-Taboada V. Performance of antinuclear antibody connective tissue disease screen. Ann N Y Acad Sci 2007;1109:322–9.10.1196/annals.1398.038Search in Google Scholar PubMed
85. Hayashi N, Kawamoto T, Mukai M, Morinobu A, Koshiba M, Kondo S, et al. Detection of antinuclear antibodies by use of an enzyme immunoassay with nuclear HEp-2 cell extract and recombinant antigens: comparison with immunofluorescence assay in 307 patients. Clin Chem 2001;47:1649–59.10.1093/clinchem/47.9.1649Search in Google Scholar
86. Gonzalez C, Garcia-Berrocal B, Perez J, Navajo JA, Herraez O, González-Buitrago JM. Laboratory screening of connective tissue diseases by a new automated ENA screening assay (EliA Symphony) in clinically defined patients. Clin Chim Acta 2005;359:109–14.10.1016/j.cccn.2005.03.042Search in Google Scholar PubMed
87. Ghillani P, Rouquette AM, Desgruelles C, Hauguel N, Le Pendeven C, Piette JC, et al. Evaluation of the LIAISON ANA screen assay for antinuclear antibody testing in autoimmune diseases. Ann N Y Acad Sci 2007;1109:407–13.10.1196/annals.1398.046Search in Google Scholar PubMed
88. Hu Y, Murphy RF. Automated interpretation of subcellular patterns from immunofluorescence microscopy. J Immunol Methods 2004;290:93–105.10.1016/j.jim.2004.04.011Search in Google Scholar PubMed
89. Glory EM, Murphy RF. Automated subcellular location determination and high throughput microscopy. Dev Cell 2007;12:7–16.10.1016/j.devcel.2006.12.007Search in Google Scholar PubMed
90. Hiemann R, Buttner T, Krieger T, Roggenbuck D, Sack U, Conrad K. Challenges of automated screening and differentiation of non-organ specific autoantibodies on HEp-2 cells. Autoimmun Rev 2009;9:17–22.10.1016/j.autrev.2009.02.033Search in Google Scholar PubMed
91. Egerer K, Roggenbuck D, Hiemann R, Weyer MG, Büttner T, Radau B, et al. Automated evaluation of autoantibodies on human epithelial-2 cells as an approach to standardize cell-based immunofluorescence tests. Arthritis Res Ther 2010;12:R40.10.1186/ar2949Search in Google Scholar PubMed PubMed Central
92. Rigon A, Buzzulini F, Soda P, Onofri L, Arcarese L, Iannello G, et al. Novel opportunities in automated classification of antinuclear antibodies on HEp-2 cells. Autoimmun Rev 2011;10:647–52.10.1016/j.autrev.2011.04.022Search in Google Scholar PubMed
93. Kivity S, Gilburd B, Agmon-Levin N, Garcia Carrasco M, Tzafrir Y, Sofer Y, et al. A novel automated indirect immunofluorescence autoantibody evaluation. Clin Rheumatol 2012;31:503–9.10.1007/s10067-011-1884-1Search in Google Scholar PubMed
94. Melegari A, Bonaguri C, Russo A, Luisita B, Trenti T, Lippi G. A comparative study on the reliability of an automated system for the evaluation of cell-based indirect immunofluorescence. Autoimmun Rev 2012. Epub ahead of print. doi:10.1016/j.autrev.2011.12.010.10.1016/j.autrev.2011.12.010Search in Google Scholar PubMed
95. Mariz H, Sato E, Barbosa SH, Rodrigues SH, Dellavance A, Andrade LE. Pattern on the antinuclear antibody-HEp-2 test is a critical parameter for discriminating antinuclear antibody-positive healthy individuals and patients with autoimmune rheumatic diseases. Arthritis Rheum 2011;63:191–200.10.1002/art.30084Search in Google Scholar PubMed
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