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Danish Multiple Sclerosis Center

Analysis in Neuroimmunology Laboratory

See detail information on the link for Rigshospitalets Labportal: http://labportal.rh.dk

  • Analysis for IgG oligoclonal bands in CSF and serum
  • Analysis for s-acetylcholine receptor antibodies in serum
  • Analysis for neutralizing IFN- antibody (RH6311) in serum
  • Analysis for the presence of Tysabri antibodies in serum
  • Analysis for the presence of Aquaporin-4 antibodies (NMO-IgG) in serum
  • Analysis of the presence of Neurofilament light polypeptide (NFL) (NPU27353)

  • 1. Analysis for IgG oligoclonal bands (NPU17076).

    The validation of oligoclonal bands in patients may be used for diagnosis of MS. By separation and identification of IgG bands in cerebrospinal fluid (CSF) and serum by use of isoelectric focusing in an agarose gel, an analysis indicates if oligoclonal antibodies are present in the CSF of the central nervous system of patients.

    The international definition of “positive for oligoclonal bands” is that at least two bands are seen in the CSF sample not present in the corresponding serum sample. If only a single band or no bands are seen in CSF as compared to serum then the result is negative. In the case of a single band only in the CSF and not in the serum, we suggest sending a new sample after 3-12 months, since there may be a development to positivity over time.

    Results of analysis:
    Samples are analysed every week, and results are given as positive or negative for oligoclonal bands. Positive results demonstrate inflammation in the CNS with production of IgG. However, this is not specific for MS, since several other diseases also may demonstrate oligoclonal bands. In MS more than 95% of the patients generate oligoclonal bands.

    The results of the analysis are sent out by internal mail at Rigshospitalet and by ordinary mail to other hospitals as soon as possible after analysis. We are in the process of coupling the local database with results to “Sundhedsplatformen)”. It may occur that we need to reanalysed samples due to required dilution of samples or uncertainty in decision of the result, and this may postpone the sending of results up to three weeks from the date of received samples. If problems arise and it will take more than three weeks to obtain a result, a letter will be sent out about this.

    Requisition
    (pdf) (click to get the requisition for print)

    On the requisition the following information is required:
    date and time of sampling (spinal fluid and serum), glass-number on both sample tubes and requisition, name and patient ID-number (cpr number), requiring department (section number at RH), and address for payment of analysis (incl. GLN-no).

    Price per analysis: 769 d.kr.

    Responsible technician: Kristian M. Nielsen

    Responsible for analysis: head of laboratory Poul Erik H. Jensen

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    2. Analysis for s-acetylcholine receptor antibodies (NPU18343).

    Autoantibodies against the acetylcholine receptor (AChR) in the neuromuscular endplate generate blockage of the neurotransmitter acetylcholine binding to the receptors on the muscular surface. This may lead to the autoimmune disease Myasthenia Gravis (MG). Demonstration of the autoantibodies has a great importance to make the diagnosis of MG, and for the follow-up of the severity of the disease.
    To help for diagnostics of MG, we perform an analysis of these autoantibodies in a serum sample from patients.
    By use of a 125I-radioreceptor-assay (ACHRAB®-Assay) kit, once a week we perform the analysis in Neuroimmunology Laboratory.

    Results of analysis:
    The results are sent out after analyses as a concentration (nmol/L) of antibodies to sAChR included a figure of the development of results (earlier analyses) over time for each patient. If the sample is first-time positive or has varied significantly in concentration, it is reanalysed the following week in a series of dilution for making a precise result.
    The results should be interpreted as followed:

    Negative result
    Unclear result
    Positive result
    0–0,2 nmol/l
    0,3–0,4 nmol/l
    >0,5 nmol/l

    In case of an unclear result, it may be suggested to analyse a new sample after 2-3 months to evaluate if it has changed to negative or positive.

    The result of the analysis is sent as soon as possible and at most within 3 weeks after receiving the sample.

    Requisition
    (pdf) (click to open requisition for print)

    Information of necessity for the analysis:
    date of sampling, sample ID-number (on both tubes and requisition), name and patient ID-number (cpr number), the addresses whereto the result of the analysis and invoice is sent, and the GLN-number is important.

    Price per analysis: 982 d.kr.

    Responsible for analysis: head of laboratory Poul Erik H. Jensen

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    3. Analysis for neutralizing IFN-ß antibodies (RH6311) (NPU27709).

    Interferon beta (IFN-ß) treatment is beneficial in relapsing-remitting and some secondary progressive MS patients. In some patients treatment results in the development of antibodies to IFN-ß that may neutralize the biological effects of treatment. Therefore it is important to analyse for the presence of IFN-ß neutralizing antibodies in the blood of MS-patients.

    The blood samples are tested by a luciferase reporter gene assay using the human cell line HT-1080 (HL-116 clone, Dr. G. Uzé, France) transfected with the firefly luciferase gene linked to an interferon-stimulated response element. When IFN-ß molecules bind to the cell surface receptors, the reporter system is activated, leading to the expression of luciferase protein in the cell. The amount of luciferase is quantified by the chemiluminescence signal generated from a specific substrate. The chemiluminescence that is measured when diluted serum and a known amount of IFN-ß is added to the cells reflects the effects of neutralizing anti-IFN-ß antibodies (NAbs) in the serum sample. The effect of neutralizing antibodies in samples testing positive for neutralizing antibodies is expressed as the tenfold reduction unit (TRU), calculated as a Kawade titre.

    Blood samples are generally tapped for analysis from the MS-patient at 6, 12, 18 and 24 months after start on IFN-ß treatment.

    Results of analysis:
    The results are given as negative or positive. Positive results are given as a titre (TRU/mL). Titres >20 TRU/mL are positive and antibodies are regarded to have influence on effect of the treatment with IFN-ß. Values <20 TRU/mL are defined as negative and indicates that the strength of antibodies (if any) do not have significant influence on the effect of treatment with IFN-ß. Titer values higher than 200 TRU/mL can be regarded as strongly positive.

    Due to a significantly fewer number of samples for analysis, we only analyse for Nab-samples every 2.weeks with start from May 2016. The result of the analysis is sent as soon as possible.

    Requisition
    (pdf) (click to open requisition for print)

    Information of necessity for the analysis:
    date of sampling, sample ID-number (on both tubes and requisition), name and patient ID-number (cpr number), the addresses whereto the result of the analysis and invoice is sent, and the GLN-number is important.

    Price per analysis: 985 d.kr (132 €) per sample.

    Responsible for analysis: head of laboratory Poul Erik H. Jensen

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    4. Analysis of the presence of Tysabri antibodies (RHB00138).

    Tysabri (Natalizumab) is a humanized, recombinant monoclonal antibody that binds to the α4-subunits of α4ß1 and α4ß7 integrin on mononuclear blood cells, and can block their binding to the adhesion molecule VCAM-1 among others on endothelial cells. The important effect of this blockage in MS-patients is that inflammatory mononuclear cells in less degree enter the central nervous system.
    Antibodies against the Tysabri product in the blood may block its binding to the integrin. Demonstration of the antibodies has a great importance for the validation of the effects of Tysabri on MS-patients.
    The validation of the beneficial effects of Tysabri on the MS-patient is perform by an ELISA-assay (developed by Biogen Idec Inc.) analysis of serum samples from patients.

    Results of analysis:
    The results are given as positive or negative for the presence of antibodies to Tysabri. The interpretation of a positive sample is that the measured antibodies in the sample are of clinical relevance (Biogen Idec).

    The result of the analysis is sent out as soon as possible and at most within 3 weeks after receiving the sample.

    Requisition
    (pdf) (click to retrieve requisition for print)

    Information of necessity for the analysis:
    date of sampling, sample ID-number (on both tubes and requisition), name and patient ID-number (cpr number), the addresses whereto the result of the analysis and invoice is sent, and the GLN-number is important.

    Price per analysis: 590 d.kr per sample (80 €).

    Responsible technician: Kristian M. Nielsen

    Responsible for analysis:
    head of laboratory Poul Erik H. Jensen

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    5. Analysis of the presence of Aquaporin-4 antibodies (NMO-IgG) (NPU27284).

    Neuromyelitis opticus (NMO), also known as Devic’s disease, is an inflammatory disease of the central nervous system characterized by severe optic neuritis and transverse myelitis, usually with a relapsing-remitting disease course. The analysis of anti-aquaporin-4 antibodies (also known as NMO-IgG) identifies circulating autoantibodies against aquaporin-4, a water channel molecule expressed on astrocytes in the central nervous system. Anti-aquaporin-4 antibodies can be detected in serum from approximately 75% of patients with NMO. Anti-aquaporin-4 antibodies can also be identified in patients with isolated, longitudinally extensive transverse myelitis or optic neuritis (NMO spectrum disease).

    Results of analysis:
    The result of the analysis is given as units/mL (U/mL), which is calculated from the amount of antibodies to aquaporin-4 in a dilution of the sample. The number of U/ml is proportional to the amount of anti-aquaporin-4 in the sample.
    The results should be interpreted as follows:
    <3 U/mL: negative for anti-aquaporin-4 antibodies.
    >=3 U/ml: positive for anti-aquaporin-4 antibodies.
    Please be aware that a maximum value may be given as a result, which is then marked by the sign > (bigger than), and shows that the actual value is higher than the one shown.

    Samples are analysed and results sent out within 3 weeks after receiving the sample.

    Analysis is performed by the use of an ELISA-kit from DLD Diagnostika, Germany. More information may be found in the publication: Jarius, S., et al., Journal of the Neurological Sciences, 320(2012), 32-37.

    Requisition
    (pdf) (click to retrieve the requisition for print)

    Information of necessity for the analysis:
    date of sampling, sample ID-number (on both tubes and requisition), name and patient ID-number (cpr number), the addresses whereto the result of the analysis and invoice is sent, and the GLN-number is important.

    Price per analysis: 878 d.kr per sample.

    Responsible for analysis: head of laboratory Poul Erik H. Jensen

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    6. Analysis of the presence of Neurofilament light polypeptide (NFL) (NPU27353).

    Results of analysis:
    Due to increase of NFL with age in humans, the results are determined by the age of the patient analysed in accordance with the table below.

    Age (years)
    <30
    30-<39
    39-<60
    >=60
    limit ng/L
    380
    560
    890
    1850

    The result of the analysis is given as units/mL (ng/L), which is calculated from the amount of NFL of the sample.
    The results should be interpreted as in the following example:
    If the result of the sample f.ex. is 600 ng/L, then the age of the patient determines if the sample is positive or negative. If the patient is 50 years old, then the sample is negative and if the patient is 30 years old, the sample is positive.

    Samples are analysed and results sent out within 3 weeks after receiving the sample.

    Analysis is performed by the use of an ELISA-kit (NF-light®) from UmanDiagnostics, Sweden.
    More information may be found in the publication: “Biomarker Evidence of Axonal Injury in Neuroasymptomatic HIV-1 Patients”, Jessen Krut J , Mellberg T , Price RW, Hagberg L, Fuchs D, Rosengren L, Nilsson S, Zetterberg H, Gisslén M, PLOS one, February 11, 2014; 9(2):e88591.

    Requisition
    (pdf) (click to retrieve the requisition for print)

    Information of necessity for the analysis:
    date of sampling, sample ID-number (on both tubes and requisition), name and patient ID-number (cpr number), the addresses whereto the result of the analysis and invoice is sent, and the GLN-number is important.

    Price per analysis: 816 d.kr per sample.

    Responsible for analysis: head of laboratory Poul Erik H. Jensen

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    Multiple Sclerosis Research Unit and Neuroimmunology Laboratory:
    Copenhagen University Hospital, Rigshospitalet, sect. 6311, Blegdamsvej 9, 2100 Copenhagen, Denmark
    Multiple Sclerosis Clinic:
    Copenhagen University Hospital, Rigshospitalet, sect. 2082, Blegdamsvej 9, 2100 Copenhagen, Denmark