A new recombinant designer antigen for autoantibody diagnostics in patients with bullous pemphigoid has been developed in a collaboration involving a substantial contribution from EUROIMMUN researchers. The new protein is much better suited for diagnostic purposes than the natural version.  From the huge BP180 molecule it contains only the pathogenetically relevant target structure –  the NC16A domain, and this as a tetramer (NC16A-4X*). On the basis of this antigenic work of art an extremely efficient ELISA test system has been developed, which exhibits a sensitivity of 90% (!) (specificity 98%!) and detects 100% of patients with gestational pemphigoid. If autoantibodies against BP230 are additionally determined, 93% of patients with bullous pemphigoid can be identified serologically.

In pemphigus foliaceus and pemphigus vulgaris autoantibodies against desmosomes are investigated. The main target antigens are the desmogleins (Dsg) 1 and 3. In pemphigus foliaceus only Dsg 1 reacts, while in pemphigus vulgaris a reaction occurs with either Dsg3 alone or  Dsg1 and Dsg3 together. For this application the use of recombinant antigens is state-of-the-art. These are synthesised at EUROIMMUN in human cell lines, in which they undergo species-faithful and thus authentic posttranslational modification. The corresponding state-of-the-art ELISAs show impressive sensitivities of 96.0% (Dsg1) and 100% (Dsg3) at specificities of 99.1% (Dsg1) and 99.6% (Dsg3).

All of these autoantibodies can also be detected by indirect immunofluorescence. Up until now the standard substrate has been tissue sections of oesophagus and tongue. However, with these substrates it is difficult to distinguish between Dsg1 and Dsg3, and patterns are often obscured due to unspecific reactions (e.g. from antibodies against keratin). Today a powerful Mosaic of specific transfected cells together with the tissue sections allows reliable diagnosis with a single test.

The test kits are equally suited to primary diagnostics, disease course assessment and therapy monitoring. The ELISAs demonstrate the following characteristics:

– CE conformative standard curve using 3 calibrators
– Positive and negative controls included in test kit
– Wide measurement range (2-200 RU/ml)
– Short, standard incubation times (30 min/30 min/15 min)
– Reagents exchangeable with those in other EUROIMMUN ELISAs
– Attractive price

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Anti-NMDA receptor encephalitis is a severe encephalopathic autoimmune disease, which affects mainly young women with ovarian teratoma, but also women without tumours, men and children. First described in 2007, it is a currently still widely underdiagnosed disease entity (J. Dalmau et al., 2008, Lancet Neurol 7(12): 1091-1098).

Diagnosis of anti-NMDA receptor encephalitis is based on the detection of highly specific autoantibodies directed against glutamate receptors of type NMDA in serum or cerebrospinal fluid. These autoantibodies are detected with high sensitivity and specificity in indirect immunofluorescence using a human recombinant cell line expressing the major target antigen (receptor subunit NR1). The determination of antibodies against glutamate receptors (type NMDA) is of high significance in patients with encephalitis where no pathogen has been detected, as well as in suspected cases of limbic encephalitis. The new cell substrate can be combined with various tissue substrates that are relevant for differential diagnostics (e.g. hippocampus and cerebellum) as BIOCHIP Mosaics, allowing the detection of further autoantibodies associated with limbic encephalitis (e.g. anti-VGKC antibodies, anti-AMPA receptor antibodies). more information

Tick bite encephalitis (TBE) may occur at any time, even outside of specified risk areas. In its most recent publication the renowned Robert Koch Institute in Berlin, Germany, points out that during tick bite season TBE infection should always be taken into account in cases of fever and headache.

EUROIMMUN offers test systems for a reliable diagnosis of anti-TBE antibodies in serum. Intrathecal synthesis can be rapidly detected using the Anti-TBE Virus ELISA for cerebrospinal fluid (CSF). EUROIMMUN also provides an avidity test. The determination of antibody avidity can be very useful in individuals with persisting anti-TBE IgM to rule out acue infection in suspected cases.

The vaccine Encepur^®, which is used most frequently for TBE immunisation, is based on antigens of the K23 virus strain. For monitoring successful immunisation, EUROIMMUN offers the only currently available Anti-TBE ELISA containing antigens of the K23 vaccine strain and at the same time providing quantification in Vienna units, giving orientation for the evaluation of results.

The determination of the HLA-B27 allele, which is present in over 90% of patients with Bechterew's disease, but is only found in 6% – 9% of the normal population, plays a key role in the diagnosis of spondylarthritides. Test systems for the determination of this genetic marker should detect all HLA-B27 subtypes, but also indicate the presence of non-disease relevant HLA-B27 subtypes.

The EUROArray HLA-B27 offers this combination. Moreover, this assay enables quick, simple and reliable determination of HLA-B27 at low costs and stands out through the following characteristics:

• An objective, fully automated evaluation and archiving system using the EUROArrayScanner – no time-consuming cell counting or subjective interpretation of gel electrophoresis and blot incubation results
• Ready-to-use PCR reagents – save additional pipetting steps
• Integrated B27 positive control – additional positive controls are not required
• Clear results for each sample – no difficult-to-interpret results caused by borderline bands such as in electrophoresis and blot methods
• No live leukocytes necessary – the samples can be collected, stored for long periods and analysed together (contrary to cytotoxicity tests, Terasaki methods or FACS analyses)
• CE-labelled for in vitro diagnostic use
• Devices available on request

Inform yourself personally about the simple handling and excellent performance of this test system, which has already exceeded the expectations of many rheumatology laboratories! more information

Do not leave reliable results in allergy diagnostics to chance! With two EUROIMMUN blot tests performed in parallel you can investigate at least 40 parameters at the same reagent costs as with ten single allergens.

Special features of allergen profile blot strips from EUROIMMUN:

– large allergen spectrum available: profiles Inhalation, Food, Atopy, Paediatrics and Insect venoms

– small sample volumes required: 200 µl serum for 40 allergens

– automation (incubation, evaluation) is the same as in other EUROIMMUN blot systems

– antibodies against CCD (cross-reactive carbohydrate determinants): an innovative extra on our allergen profile blot strips is the determination of CCD reactivity, which is a frequent cause of positive in vitro reactions with missing clinical correlate.

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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).

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 research 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

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.

The detection of agent-specific IgM antibodies in patient serum is generally considered to confirm an acute infection. However, IgM antibodies can be found only temporarily during the course of the disease and do not allow identification of all acute infections. In other cases, the IgM persists for months or years after recovery. If the IgM investigation was not helpful, a modern diagnostic laboratory nowadays is also able to serologically diagnose a fresh infection by determining the avidity of the specific IgG: At first contact with an unknown infectious agent the organism starts to produce antibodies with a weak affinity. During the course of the disease, however, the adaptation of the B-lymphocytes to the agent becomes more accurate, inceasing the initially low avidity of the specific IgG by several powers. The determination of high-avidity IgG antibodies at first testing is therefore a good indication that the actual infection took place some time ago, whereas the presence of low-avidity IgG confirms a fresh infection.

• 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

• One for all: fully automated processing of all EUROIMMUN ELISA – autoimmune diagnostics, infectious serology and allergology – with only one system.
• Flexibility for your benefit: open system with more than 800 validated EUROIMMUN parameters for serum, plasma and cerebrospinal fluid, over 50 or 30 parameters in parallel.
• Convenient, simple and reliable operation: e.g. by scanning QC certificates using a 2D-hand barcode scanner – ready-for-use reagents and preprogrammed assay protocols enable  you to start immediately.
• Capacity and throughput: quick loading and efficient time management allow processing of up to 70 or 50 tests per hour – up to 7 or 3 plates, 180 or 144 samples at the start of a run.
• Security for patients: validated test systems and the comprehensive safety kit provide the basis for reliable diagnostics.
• Reliability and service: instruments and reagents from one manufacturer, quick and targeted support from our personnel for a smooth workflow in your laboratory.

Product information Analyzer I

Product information Analyzer I-2P

For the completion of the diagnostic spectrum, EUROIMMUN has extended its product range in nuclear medical diagnostistics by high-performance radioimmunoassays for the following indications:

• Thyroid diseases: autoantibodies against thyroperoxidase (TPO), thyroglobulin (TG), TSH receptor (TRAb); Antigen: thyroglobulin (TG); Hormone determinations: free trijodthyronine (FT3), free thyroxine (FT4), thyrotropin (TSH), calcitonin
• Diabetes mellitus: autoantibodies against glutamic acid decarboxylase (GAD), tyrosine phosphatase (IA2), insulin (IAA)
• Myasthenia syndrome: autoantibodies against acetylcholine receptors (ACHR), P/Q calcium channels (VGCC)
• Systemic lupus erythematosus: autoantibodies against double-stranded DNA (dsDNS).