ravo    


PNS14 DIVER and PNS14 Line Assay

PNS14 DIVER and PNS14 Line Assay
more...
Soon available

MAG DIVER and Gangliosid Line Assay
more...

Immuno-(Dot)-Blot for the detection of the paraneoplastic autoantibodies anti-HuD, anti-Yo, anti-Ri, anti-CV2 (CRMP5),
anti-Amphiphysin, anti-Ma1 and anti-Ma2


The assessment of antineuronal antibodies (anti-Hu, anti-Yo, anti-Ri, anti-CV2 (anti-CRMP5), anti-Amphiphysin, anti-Ma1 and anti-Ma2 IgG-antibodies) is playing an increasing role in the differential diagnosis of sensory neuropathy, cerebellar ataxia and encephalomyelitis. Demonstration of these autoantibodies in the presence of paraneoplastic neurological symptoms provides strong diagnostic evidence of a - possibly occult - neoplasm. In two-thirds of the cases, a paraneoplastic neurological syndrome precedes the discovery of an underlying neoplasm by several years. Thus, demonstration of paraneo-plastic antineuronal autoantibodies may lead to an early discovery of cancer (see table).

Generally, these autoantibodies have been identified by immunohistochemical techniques. Due to problems with specificity a positive result in immunohistochemistry needs to be confirmed by an immunoblot, employing crude extracts from neuronal tissue as antigen. For example the reactivity of anti-Ro-antibodies, associated with Sjögren's-Syndrome, with protein bands resembling anti-HuD-immunoreactivity in crude extracts from neuronal tissue has been described. Immunohistochemistry is also laborious and requires a high degree of experience for reliable interpretation.

Figure 1:

1      positive control
2      negative serum sample
3-24 different positive serum samples


Sensitivity and Specificity

47 positive serum samples, which were confirmed by IFT and other tests, were tested. Among these were 22 HuD-, 6 Yo-, 8 Ri-, 4 CV2-, 3 Amphiphysin- and 4 Ma2-positiv serum samples. All these samples were clearly identified as positive. To test spezificity, 46 serum samples of healthy persons, as well as patients showing other neurological symptoms, were tested. These samples showed no reaction with the antigens.

Table: Paraneoplastic neurological syndromes Most frequently associated tumors
Anti-Hu-Antibodies(ANNA-1)
  • Sensory and autonomic neuropathy
  • Cerebellar ataxia
  • Encephalomyelitis
  • Limbic Encephalitis
Small-cell-lung
cancer
Non-small-cell
lung cancer
Extrapulmonary
small cell cancer
Anti-Yo-Antibodies(Purkinje-cell-antigen)
  • Cerebellar ataxia
Breast cancer
Ovarian cancer
Uterus cancer
Anti-Ri-Antibodies(ANNA-2, anti-Nova-1)
  • Brainstem encephalitis (incl. Opsoclonus-Myoclonus-Syndrome)
  • Cerebellar ataxia
Breast cancer
Small-cell-lung cancer
Medullary carcinoma of the thyroid gland
Anti-CV2-(CRMP5-) Antibodies
  • Sensory and sensorimotor neuropathy
  • Encephalomyelitis
  • Cerebellar ataxia
  • Limbic Encephalitis
  • Autonomic neuropathy
  • Chorea
Small-cell-lung
cancer
Thymom
Anti-Amphiphysin-Antibodies
  • Stiff-person-syndrom
  • Various symptoms
Breast cancer
Small-cell-lung cancer
Anti-Ma1 and Anti-Ma2- (Ta-) Antibodies
  • Limbic Encephalitis
  • Brainstem encephalitis*
  • Cerebellar ataxia*
Testicular cancer
Lung-cancers

* Brainstem encephalitis and cerebellar ataxia usually associated with tumors other than testicular and immunoreactivity against Ma2 and Ma1 proteins

Short description of test performance:

  • Serum samples are diluted 1:2,000 in ready to use sample dilution buffer.

  • Incubate the strips with 2 ml of the diluted serum specimen for 60 minutes at room temperature on a rocking table.

  • wash five times with diluted wash buffer.

  • Add 2 ml alkaline phosphatase IgG conjugate, ready to use, per strip.

  • Incubate for 30 minutes at room temperature on a rocking table.

  • wash five times with diluted wash buffer.

  • Incubate each strip in 2 ml ready to use substrate-solution.

  • Incubate for 25 minutes at room temperature until the bands become clearly visible. See control scan for comparison.

  • Transfer the strips to distilled water to stop the reaction. Put the strips onto filter paper and let them dry. Store the strips in the dark.

    back
ravo Diagnostika, Oltmannsstr. 5, D-79100 Freiburg
phone +49 761 40 74 88, fax +49 761 40 74 77, email