In ANCA diagnostics clincians repeatedly come across sera that show an ambiguous result in indirect immunofluorescence (IIF) or for which the confirmatory ELISA is not sensitive enough. In these cases, a reliable interpretation of the overall result is rendered impossible by discrepant data. EUROIMMUN now offers several innovations to solve these problems:

1.) In indirect immunofluorescence we have supplemented our Granulocyte Mosaic with further very useful BIOCHIPs. HEp-2 cells coated with granulocytes enable immediate differentiation between ANCA and ANA, BIOCHIPs with PR3 or MPO antigen dots (EUROPLUS) provide additional information for reliable interpretation of results. In daily practice, this means time savings in evaluation and establishment of diagnosis.

2.) In the diagnosis of Wegener’s granulomatosis, autoantibodies against proteinase 3 can be detected with yet unsurpassed sensitivity by ELISA using a new human designer antigen (see poster). In a clinical study carried out by the ANCA reference centre of the University of Maastricht, the new EUROIMMUN Anti-PR3-hn-hr ELISA showed an impressive hit rate in laboratory routine (Damoiseaux et al., Ann Rheum Dis, 2008 Mar 28, Epub ahead of print). More Information

Antibodies against double-stranded DNA (dsDNA) are the main focus in the serological diagnosis of systemic lupus erythematosus (SLE). These antibodies can be found in 60% to 90% of patients, depending on the activity of the disease. Antibodies against nucleosomes are also an exclusive marker for SLE, provided that they are determined using a perfect test system whose target antigen must be free of histone H1, Scl-70 and other non-histone proteins.

Using an innovative biochemical preparation, researchers at EUROIMMUN AG have developed a new test system that exceeds the diagnostic quality criteria of all conventional Anti-dsDNA ELISA by far (Anti-dsDNA-NcX ELISA). The secret of the innovation lies in the use of highly purified nucleosomes as the new linking substance. Since nucleosomes have a strong adhesive ability, even the smallest concentration of these is highly suited to couple isolated dsDNA to the surface of a microplate well. Poly-L-lysine and potamine sulphate have fallen into disuse, many false positive results are prevented, the specificity of the ELISA amounts to values which equal those of the indirect immunofluorescence test using Crithidia luciliae! In a clinical comparison study using 378 samples from patients with rheumatic diseases (209 of these with SLE) the Anti-dsDNA-NcX ELISA clearly demonstrated its superiority over the Anti-dsDNA RIA (Farr assay), showing an 8% higher sensitivity.

The Anti-dsDNS-NcX-ELISA can be performed manually or fully automatically (EUROIMMUN Analyzer I). Other competent test systems such as the Farr assay and the indirect immunofluorescence test (substrate Crithidia luciliae) continue to be of importance in the clarification of discrepant serological and clinical results. More information

Our hard working research groups are constantly engaged in developing the most state-of-the-art test systems. Diagnosis of rheumatic diseases is among the fields which are most important to us. In the following, we would like to present some of the test systems EUROIMMUN offers for the serological diagnosis of rheumatic diseases:

As a pioneer in the worldwide introduction of anti-CCP diagnostics EUROIMMUN is among the small number of companies which are able to offer an Anti-CCP ELISA in accordance with the patent of Walter van Venrooij et al. With respect to disease specificity the EUROIMMUN test system is unsurpassed by any other commercial test (N. Bizzaro et al., 2006, 5th International Congress on Autoimmunity, Sorrent, Italy): Some manufacturers use suboptimal antigens to avoid the patent. The Anti-CCP ELISA, however, ranks first in serological diagnostics of rheumatoid arthritis.

The Anti-dsDNA-NcX ELISA is a new milestone in the diagnosis of systemic lupus erythematosus (SLE). Today, DNA is coupled to the surface by means of highly purified, wafer-thinly applied nucleosomes as linker substrate and not, as in the past, by polylysine or protamine sulphate, which sometimes cause unspecific reactions. Due to this new coating technique the new Anti-dsDNA-NcX ELISA is much more sensitive than conventional anti-dsDNA tests: In the investigation of a large SLE panel (R. Biesen et al., 2008, Lupus 17(5): 506-507) the test achieved a sensitivity of 60.8% at a specificity of 98% (Anti-dsDNA RIA by Farr: 53.1%, conventional Anti-dsDNA ELISA: 35.4%, C. luciliae IIFT: 27.4%).

The product "EUROLINE" by EUROIMMUN encompasses a variety of line blot systems which are particularly useful for the determination of antibody profiles in autoimmune diagnostics, infectious serology and allergology. EUROIMMUN researchers examined the disease specificity of antibodies against Ro-52 using these EUROLINE profiles: In cooperation with the Charité, Berlin, Germany, and the You-An Hospital, Beijing, China, it was confirmed that antibodies against Ro-52 are not associated with a specific disease but occur in a large number of different autoimmune and even infectious diseases (Meyer et al., 2008, Annals of the Rheumatic Diseases 67 (Suppl. II): 146). Therefore, we recommend not to use any test systems based on a mixture of SS-A (60 kDa) and Ro-52 for the determination of antibodies against SS-A in the diagnosis of Sjögren's syndrome and SLE since these lead to unspecific reactions!

This microscope is specifically tailored to the requirements of indirect immunofluorescence. The labourious conventional mercury vapour lamp has been replaced by the stunningly simple EUROStar Bluelight system. Now available with electronically controlled constant light intensity. more information

• For all EUROIMMUN blot systems: EUROLINE, EUROLINE-WB, Westernblot.
• For all areas: autoimmune diagnostics, infectious serology and allergology.
• EUROBlotCamera: digitalisation of strips while in the incubation tray.
• EUROBlotScanner: digitalisation of strips using flatbed scanner.
• Fully automated identification, quantitation and assignment of bands.
• Option to modify results (changes are automatically documented).
• Complete results obtained just a few minutes after finishing the incubation.
• Fully automated administration and documentation of extensive individual data.
• Electronic archiving of all images and data (avoids the need to store potentially infectious blot strips).

• Standardisation of immunoblot strips – higher precision
  and reproducibility
• Automatisation of all EUROIMMUN immunoblot strips (EUROLINE, EUROLINE-WB, Westernblot)
• Over 65 validated parameters available (16 autoantibody, 28 infectious and 21 allergy parameters)
• Up to 30 strips per test run
• Easy operation
• Combination of different conjugates/tests in one run
• Walk-away function – fully automated from the start to the end of processing following loading
• Compatible with modern evaluation systems such as EUROBlotCamera and EUROLineScan

Automation of all EUROIMMUN ELISA (over 900 validated parameters: 60 autoantibody, 100 infection and 740 allergy parameters). 7 Microtiter plates can be loaded at the start, unusally short loading time. High convenience through use of barcode identification. Reloading function for patient samples, microtiter plates, reagents and disposable tips. "Walk-away function" – fully automated from the start to the end of processing following loading. Own processor, independent of the external IT system, guaranteeing against risk of system crashes. Bidirectional online connection to the laboratory computer system. more information

Autoimmune hepatitis (AIH) is an immune-mediated chronic inflammation of the liver. In order to secure diagnosis and to rule out combined liver disease (overlap syndrome) it is essential to differentiate AIH from alcohol- or drug-induced cirrhosis and other chronic liver diseases.

The determination of antibodies against F-actin is of particular importance in the diagnosis of autoimmune hepatitis. In contrast to other ASMA, antibodies against F-actin represent a highly specific marker for AIH type I. They cannot be determined using ELISA or Westernblot since they are directed against conformational epitopes that are only preserved in frozen sections. As there are no monospecific tests available, a clear result in IIFT is very important.

For this reason, EUROIMMUN has developed cell line VSM47 (vascular smooth muscle), which allows the quick and reliable differentiation between micro-filamentous (MF) fluorescence patterns and non-MF patterns. The use of cell line VSM47 facilitates and confirms diagnosis, thus representing a new standard in AIH diagnostics.

See also: Villalta et al., Autoimmunity 2008

To our knowledge, up until now no one has succeeded in making OspC (25kDa) - the IgM reactive antigen that is particularly important for the detection of a fresh Borrelia infection - in recombinant form in the quality needed for diagnostics. Expression systems are not (yet) capable of generating OspC epitopes in the same way as cultured Borrelia. Attempts have been made to increase the low sensitivity of recombinant OspC by applying more antigen, but this just leads to an unacceptable loss in specificity: every fifth blood donor reacts positively in blot tests - with the result that clinicians generally turn a blind eye regarding these bands and only count strong reactions. But many true positive reactions fall through, for whose sake the test is performed. For us that is not acceptable.

Therefore, as a confirmatory test in two-stage borreliosis diagnostics EUROIMMUN has always favoured classic Westernblots, which are based on Borrelia cultures. Recombinant antigens are then applied additionally (EUROLINE-WB), when they are at least on a par with the native antigens or cannot be expressed in Borrelia cultures (e.g. VlsE: Borrelia only form this anitgen in the host organism when the corresponding immunological reaction is triggered). Many customers are delighted with our Anti-Borrelia EUROLINE-WB, especially since evaluation can now be peformed automatically and reations archived electronically using EUROLineScan.

In the meantime further Borrelia antigens have been identified (several of them in our own reaserach laboratories), which are produced in insufficient quantities in cultured Borrelia, or membrane lipids, which are not presented in classic Westernblots. Moreover, we have generated completely new recombinant designer antigens - these are described in the enclosed information sheet. The latest advance is a state-of-the-art blot system, the Anti-Borrelia EUROLINE-RN-AT, in which recombinant and native antigens are placed next to each other. With the new product we are reaching out to users who wish to evaluate their Borrelia blots visually, but prefer line blots because the identification of Westernblot bands is too complicated. This blot can also be evaluated completely automatically. Reactions are scanned in the incubation bath, and interpretation of results and archiving for 48 blot strips takes less than one minute. The broadening of the antigen spectrum allows more rare antibody constellations to be detected, which helps increase the sensitivity. So, for example, native (specifically reacting) OspC from B. afzelii is supplemented with OspC from the genospecies B. burgdorferi and B. garinii, and in fresh infections 10% more positive IgM reactions are detected.

As well as the new Anti-Borrelia EUROLINE-RN-AT EUROIMMUN offers a complete package for Borrelia diagnostics that leaves no wishes unfulfilled. This is based on international guidelines for two-strep diagnosis of Borrelia (DGHM; RKI, Berlin, Germany; CDC, Atlanta, USA) and for CSF diagnostics. All reagents are CE certified and there are comprehensive automation concepts for all test systems – we are happy to provide extensive validation data on request. More information

* Includes ready-to-use, colour-coded controls for checking incubation/calculation of results

Increased tourism to exotic destinations is providing a challenge for doctors: for example, in Germany four million people travel to tropical or subtropical countries every year, and almost one in five of them returns with diarrhoea or fever. A typical tropical infection can be detected in around 15% of these. Imported infections with viruses such as Dengue also represent a growing problem. The Robert Koch Institute estimates several hundred cases per year in Germany with a high number of unidentified cases. Since the main symptoms of many tropical febrile diseases are similar and their endemic areas overlap, the specific detection of pathogens and antibodies often helps to clarify ambiguous results and to exclude dangerous infectious diseases. EUROIMMUN has developed an extensive range of indirect immunofluorescence products to enable laboratories to perform reliable, standardised serological diagnostics.

Test systems fo the following pathogens are already available (singe tests or BIOCHIP Mosaics^™ in user-defined combinations): SARS corona virus, Dengue virus (types 1-4), West Nile virus, Yellow fever virus, Japanese encephalitis virus, sandfly fever virus (types Sicilian, Naples, Toscana and Cyprus), Hantavirus (types Hantaan, Puumala, Seoul, Dobrava, Saaremaa, Sin Nombre and Andes) and Chikungunya virus. EUROIMMUN also offers ELISA for the determination of antibodies against SARS corona virus, Dengue virus type 2 and West Nile virus. Products are certified according to the IVD guideline 98/79/EC (CE registration). Test systems are evaluated by scientific studies performed in collaboration with renowned institutes.  More information