Order Code DYS2 Dysautonomia, Autoimmune/Paraneoplastic Evaluation, Serum
Ordering Guidance
Multiple neurological phenotype-specific autoimmune/paraneoplastic evaluations are available. For more information as well as phenotype-specific testing options, refer to Autoimmune Neurology Test Ordering Guide.
When more than one evaluation is ordered on the same order number, the duplicate test will be canceled.
For a list of antibodies performed with each evaluation, see Autoimmune Neurology Antibody Matrix.
This test should not be requested for patients who have recently received radioisotopes, therapeutically or diagnostically, because of potential assay interference. The specific waiting period before specimen collection will depend on the isotope administered, the dose given, and the clearance rate in the individual patient. Specimens will be screened for radioactivity prior to analysis. Radioactive specimens received in the laboratory will be held 1 week and assayed if sufficiently decayed or canceled if radioactivity remains.
Necessary Information
Provide the following information:
-Relevant clinical information
-Ordering provider name, phone number, mailing address, and e-mail address
Specimen Required
Patient Preparation: For optimal antibody detection, specimen collection is recommended before initiation of immunosuppressant medication or intravenous immunoglobulin treatment.
Supplies: Sarstedt Aliquot Tube, 5 mL (T914)
Collection Container/Tube:
Preferred: Red top
Acceptable: Serum gel
Submission Container/Tube: Plastic vial
Specimen Volume: 4 mL
Collection Instructions: Centrifuge and aliquot serum into a plastic vial.
Forms
If not ordering electronically, complete, print, and send a Neurology Specialty Testing Client Test Request (T732) with the specimen.
Useful For
Investigating idiopathic dysautonomic symptoms
Directing a focused search for cancer in patients with idiopathic dysautonomia
Investigating autonomic symptoms that appear in the course or wake of cancer therapy and are not explainable by recurrent cancer or metastasis (detection of autoantibodies in this profile helps differentiate autoimmune dysautonomia from the effects of chemotherapy)
Profile Information
Test ID | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
ADEI | Dysautonomia, Interpretation, S | No | Yes |
GANG | AChR Ganglionic Neuronal Ab, S | No | Yes |
ANN1S | Anti-Neuronal Nuclear Ab, Type 1 | No | Yes |
APBIS | AP3B2 IFA, S | No | Yes |
CRMS | CRMP-5-IgG, S | No | Yes |
CS2CS | CASPR2-IgG CBA, S | No | Yes |
DPPCS | DPPX Ab CBA, S | No | Yes |
LG1CS | LGI1-IgG CBA, S | No | Yes |
PCAB2 | Purkinje Cell Cytoplasmic Ab Type 2 | No | Yes |
Reflex Tests
Test ID | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
AN1BS | ANNA-1 Immunoblot, S | No | No |
AN2BS | ANNA-2 Immunoblot, S | No | No |
DPPTS | DPPX Ab IFA Titer, S | No | No |
AN1TS | ANNA-1 Titer, S | No | No |
APBCS | AP3B2 CBA, S | No | No |
APBTS | AP3B2 IFA Titer, S | No | No |
CRMTS | CRMP-5-IgG Titer, S | No | No |
PC2TS | PCA-2 Titer, S | No | No |
CRMWS | CRMP-5-IgG Western Blot, S | Yes | No |
Testing Algorithm
If the indirect immunofluorescence assay (IFA) patterns suggest collapsin response-mediator protein (CRMP)-5-IgG, then the CRMP-5-IgG IFA titer and CRMP-5-IgG Western blot will be performed at an additional charge.
If the IFA pattern suggests antineuronal nuclear antibody type 1 (ANNA-1), then the ANNA-1 immunoblot, ANNA-1 IFA titer, and ANNA-2 immunoblot will be performed at an additional charge.
If the IFA pattern suggests adaptor protein 3 beta 2 (AP3B2) antibody, then the AP3B2 cell-binding assay (CBA) and AP3B2 IFA titer will be performed at an additional charge.
If the dipeptidyl-peptidase-like protein-6 antibody (DPPX) antibody CBA result is positive, then DPPX IFA titer will be performed at an additional charge.
If the IFA pattern suggests Purkinje cytoplasmic antibody type 2 (PCA-2), then the PCA-2 titer is performed at an additional charge.
For more information see Autoimmune/Paraneoplastic Dysautonomia Evaluation Algorithm.
Method Name
ANN1S, AN1TS, APBIS, APBTS, DPPTS, PCAB2, PC2TS, CRMS, CRMTS: Indirect Immunofluorescence Assay (IFA)
APBCS, CS2CS, DPPCS, LG1CS: Cell Binding Assay (CBA)
CRMWS: Western Blot (WB)
AN1BS, AN2BS: Immunoblot (IB)
GANG: Radioimmunoassay (RIA)
ADEI: Medical Interpretation
Reporting Name
Dysautonomia, Autoimm/Paraneo, SSpecimen Type
SerumSpecimen Minimum Volume
2.5 mL
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Serum | Refrigerated (preferred) | 28 days | |
Frozen | 28 days | ||
Ambient | 72 hours |
Reject Due To
Gross hemolysis | Reject |
Gross lipemia | Reject |
Gross icterus | Reject |
Reference Values
Test ID |
Reporting name |
Methodology* |
Reference value |
ADEI |
Dysautonomia, Interpretation, S |
Medical interpretation |
Interpretive report |
GANG |
AChR Ganglionic Neuronal Ab, S |
RIA |
≤0.02 nmol/L |
ANN1S |
Anti-Neuronal Nuclear Ab, Type 1 |
IFA |
Negative |
APBIS |
AP3B2 IFA, S |
IFA |
Negative |
CS2CS |
CASPR2-IgG CBA, S |
CBA |
Negative |
CRMS |
CRMP-5-IgG, S |
IFA |
Negative |
DPPCS |
DPPX Ab CBA, S |
CBA |
Negative |
LG1CS |
LGI1-IgG CBA, S |
CBA |
Negative |
PCAB2 |
Purkinje Cell Cytoplasmic Ab Type 2 |
IFA |
Negative |
Reflex Information:
Test ID |
Reporting name |
Methodology* |
Reference value |
AN1BS |
ANNA-1 Immunoblot, S |
IB |
Negative |
AN1TS |
ANNA-1 Titer, S |
IFA |
<1:240 |
AN2BS |
ANNA-2 Immunoblot, S |
IB |
Negative |
APBCS |
AP3B2 CBA, S |
CBA |
Negative |
APBTS |
AP3B2 IFA Titer, S |
IFA |
<1:240 |
CRMTS |
CRMP-5-IgG Titer, S |
IFA |
<1:240 |
CRMWS |
CRMP-5-IgG Western Blot, S |
WB |
Negative |
DPPTS |
DPPX Ab IFA Titer, S |
IFA |
<1:240 |
PC2TS |
PCA-2 Titer, S |
IFA |
<1:240 |
*Methodology abbreviations:
Immunofluorescence assay (IFA)
Cell-binding assay (CBA)
Western blot (WB)
Radioimmunoassay (RIA)
Immunoblot (IB)
Neuron-restricted patterns of IgG staining that do not fulfill criteria for ANNA-1, CRMP-5-IgG, or PCA-2 may be reported as "unclassified anti-neuronal IgG." Complex patterns that include nonneuronal elements may be reported as "uninterpretable."
Note: CRMP-5 titers lower than 1:240 are detectable by recombinant CRMP-5 Western blot analysis. CRMP-5 Western blot analysis will be done on request on stored serum (held 4 weeks). This supplemental testing is recommended in cases of chorea, vision loss, cranial neuropathy, and myelopathy. Call 800-533-1710 to request CRMP-5 Western blot.
Method Description
Cell Binding Assay:
Patient specimen is applied to a composite slide containing transfected and nontransfected HEK-293 cells. After incubation and washing, fluorescein-conjugated goat-antihuman IgG is applied to detect the presence of patient IgG binding.(Package insert: IIFT: Neurology Mosaics, Instructions for the indirect immunofluorescence test. EUROIMMUN; FA_112d-1_A_UK_C13, 02/2019)
Indirect Immunofluorescence Assay:
The patient's sample is tested by a standardized immunofluorescence assay that uses a composite frozen section of mouse cerebellum, kidney, and gut tissues. After incubation with sample and washing, fluorescein-conjugated goat-antihuman IgG is applied. Neuron-specific autoantibodies are identified by their characteristic fluorescence staining patterns. Samples that are scored positive for any neuronal nuclear or cytoplasmic autoantibody are titrated to an endpoint. Interference by coexisting non-neuron-specific autoantibodies can usually be eliminated by serologic absorption.(Honorat JA, Komorowski L, Josephs KA, et al. IgLON5 antibody: neurological accompaniments and outcomes in 20 patients. Neurol Neuroimmunol Neuroinflamm 2017;4(5):e385. doi:10.1212/NXI.0000000000000385)
Radioimmunoassay:
(125)I-labeled recombinant human antigens or labeled receptors are incubated with patient specimen. After incubation, anti-human IgG is added to form an immunoprecipitate. The amount of (125)I-labeled antigen in the immunoprecipitate is measured using a gamma-counter. The amount of gamma emission in the precipitate is proportional to the amount of antigen-specific IgG in the specimen. Results are reported as units of precipitated antigen (nmol) per liter of patient sample.(Griesmann GE, Kryzer TJ, Lennon VA. Autoantibody profiles of myasthenia gravis and Lambert-Eaton myasthenic syndrome. In: Rose NR, Hamilton RG, eds. Manual of Clinical and Laboratory Immunology. 6th ed. ASM Press; 2002:1005-1012; Walikonis JE, Lennon V.A Radioimmunoassay for glutamic acid decarboxylase [GAD65] autoantibodies as a diagnostic aid for stiff-man syndrome and a correlate of susceptibility to type 1 diabetes mellitus. Mayo Clin Proc. 1998 December;73[12]:1161-1166; Jones AL, Flanagan EP, Pittock SJ, et al. Responses to and outcomes of treatment of autoimmune cerebellar ataxia in adults. JAMA Neurol. 2015;72[11]:1304-1312. doi:10.1001/jamaneurol.2015.2378)
Immunoblot:
All steps are performed at room temperature (18 to 28° C) utilizing the EUROBlot One instrument. Diluted patient serum (1:101) is added to test strips (strips containing recombinant antigen manufactured and purified using biochemical methods) in individual channels and incubated for 30 minutes. Positive samples will bind to the purified recombinant antigen and negative samples will not bind. Strips are washed to remove unbound serum antibodies and then incubated with anti-human IgG antibodies (alkaline phosphatase-labelled) for 30 minutes. The strips are again washed to remove unbound anti-human IgG antibodies and nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolylphosphate substrate is added. Alkaline phosphatase enzyme converts the soluble substrate into a colored insoluble product on the membrane to produces a black band. Strips are digitized via picture capture on the EUROBlot One instrument and evaluated with the EUROLineScan software.(O'Connor K, Waters P, Komorowski L, et al. GABAA receptor autoimmunity: A multicenter experience. Neurol Neuroimmunol Neuroinflamm. 2019;6[3]:e552. doi:10.1212/NXI.0000000000000552)
Western Blot:
Neuronal antigens extracted aqueously from adult rat cerebellum, full-length recombinant human collapsin response-mediator protein-5 (CRMP-5), or full-length recombinant human amphiphysin protein is denatured, reduced, and separated by electrophoresis on 10% polyacrylamide gel. IgG is detected autoradiographically by enhanced chemiluminescence.(Yu Z, Kryzer TJ, Griesmann GE, Kim K, Benarroch EE, Lennon VA. CRMP-5 neuronal autoantibody: marker of lung cancer and thymoma-related autoimmunity. Ann Neurol. 2001 February;49[2]:146-154; Dubey D, Jitprapaikulsan J, Bi H, et al. Amphiphysin-IgG autoimmune neuropathy: A recognizable clinicopathologic syndrome. Neurology. 2019;93(20):e1873-e1880. doi:10.1212/WNL.0000000000008472)
Day(s) Performed
Performing Laboratory
Mayo Clinic Laboratories in RochesterCPT Code Information
83519
86255 x 7
84182 AN1BS (if appropriate)
86256 AN1TS (if appropriate)
84182 AN2BS (if appropriate)
86255 APBCS (if appropriate)
86256 APBTS (if appropriate)
86256 CRMTS (if appropriate)
84182 CRMWS (if appropriate)
86256 DPPTS (if appropriate)
86256 PC2TS (if appropriate)