title: Multiplex serology of paraneoplastic antineuronal antibodies.
abstract: Paraneoplastic neurological syndromes (PNS) are devastating neurological disorders secondary to cancer, associated with onconeural autoantibodies. Such antibodies are directed against neuronal antigens aberrantly expressed by the tumor. The detection of onconeural antibodies in a patient is extremely important in diagnosing a neurological syndrome as paraneoplastic (70% is not yet known to have cancer) and in directing the search for the underlying neoplasm. At present six onconeural antibodies are considered 'well characterized' and recognize the antigens HuD, CDR62 (Yo), amphiphysin, CRMP-5 (CV2), NOVA-1 (Ri), and Ma2. The gold standard of detection is the characteristic immunohistochemical staining pattern on brain tissue sections combined with confirmation by immunoblotting using recombinant purified proteins. Since all six onconeural antibodies are usually analyzed simultaneously and objective cut-off values for these analyses are warranted, we developed a multiplex assay based on Luminex technology. Reaction of serial dilutions of six onconeural standard sera with microsphere-bound antigens showed lower limits of detection than with Western blotting. Using the six standard sera at a dilution of 1:200, the average within-run coefficient of variation (CV) was 4% (range 1.9-7.3%). The average between-run within-day CV was 5.1% (range 2.9-6.7%) while the average between-day CV was 8.1% (range 2.8-11.6%). The shelf-life of the antigen coupled microspheres was at least two months. The sensitivity of the multiplex assay ranged from 83% (Ri) to 100% (Yo, amphiphysin, CV2) and the specificity from 96% (CV2) to 100% (Ri). In conclusion, Luminex-based multiplex serology is highly reproducible with high sensitivity and specificity for the detection of onconeural antibodies. Conventional immunoblotting for diagnosis of onconeural antibodies in the setting of a routine laboratory may be replaced by this novel, robust technology.
authors.affiliation: Department of Neurology, Erasmus MC, Rotterdam, Netherlands. p.maat@erasmusmc.nl
authors.forename: Peter
authors.initials: P
authors.lastname: Maat
authors.forename: Eric
authors.initials: E
authors.lastname: Brouwer
authors.forename: Esther
authors.initials: E
authors.lastname: Hulsenboom
authors.forename: Martijn
authors.initials: M
authors.lastname: VanDuijn
authors.forename: Marco W J
authors.initials: MW
authors.lastname: Schreurs
authors.forename: Herbert
authors.initials: H
authors.lastname: Hooijkaas
authors.forename: Peter A E Sillevis
authors.initials: PA
authors.lastname: Smitt
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000912:Antibodies, Neoplasm / Q000097:blood*; D000951:Antigens, Neoplasm / Q000276:immunology; D001323:Autoantibodies / Q000097:blood*; D015153:Blotting, Western; D051959:ELAV Proteins / Q000276:immunology; D000067779:ELAV-Like Protein 4; D005260:Female; D006801:Humans; D006867:Hydrolases; D007150:Immunohistochemistry; D057230:Limit of Detection; D008297:Male; D008869:Microtubule-Associated Proteins; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000276:immunology*; D000090682:Neuro-Oncological Ventral Antigen; D020361:Paraneoplastic Syndromes, Nervous System / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D011237:Predictive Value of Tests; D016601:RNA-Binding Proteins / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D001323:Autoantibodies; C095184:CDR2 protein, human; D051959:ELAV Proteins; D000067779:ELAV-Like Protein 4; C498010:ELAVL4 protein, human; C119249:Ma2 antigen; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D000090682:Neuro-Oncological Ventral Antigen; D016601:RNA-Binding Proteins; C075637:amphiphysin; C415087:DPYSL5 protein, human; D006867:Hydrolases