CDSP - Performance: Celiac Disease Serology Cascade

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Test ID: CDSP    
Celiac Disease Serology Cascade

Method Description Describes how the test is performed and provides a method-specific reference

Immunoglobulin A (IgA):

Total IgA levels are measured by immunonephelometry. Rabbit antisera that specifically recognizes human IgA is added to the patient sample. Immune complexes form between the human IgA and the rabbit immunoglobulins; the immune complexes scatter light that is passed through the sample. The intensity of the scattered light is related to the amount of human IgA in the sample. The concentration of IgA in the sample is calculated from a multipoint calibration curve.(Instruction manual: Behring Nephelometer II Operations. Dade Behring, Inc. Newark, DE 19714)

 

Tissue Transglutaminase (tTG) Antibody, IgA/IgG:

IgA and IgG antibodies to tTG are detected by enzyme-linked immunosorbent assay (ELISA). Recombinant human tTG antigen expressed in Escherichia coli is adsorbed to wells of a microtiter plate under conditions that preserve the native state of the antigen. Diluted patient sera are added to the microtiter plate wells under conditions that allow binding of the antibodies to the tTG antigen. Unbound sample constituents are washed away and horseradish peroxidase (HRP)-labeled antihuman IgA or IgG antibody conjugate is added to each well. After a second incubation, unbound HRP-label is removed and bound conjugate is detected by adding tetramethylbenzidine (TMB) chromogenic substrate. After a final incubation, colored product is measured spectrophotometrically; the absorbance of the patient sample is compared to the positive calibrator. The absorbance is directly proportional to the level of IgA or IgG antibodies to tTG, which is expressed in arbitrary units.(Instruction manual: QUANTA Lite R h-tTG IgA and IgG. Inova Diagnostics, Inc. San Diego, CA, 92131)

 

Gliadin (Deamidated) Antibody, IgA/IgG:

IgA and IgG antibodies to deamidated gliadin peptides are detected by ELISA. Purified peptides are adsorbed to wells of a microtiter plate under conditions that preserve the native state of the antigen. Diluted patient sera are added to the microtiter plate wells under conditions that allow binding of the antibodies to the deamidated gliadin peptides. Unbound sample constituents are washed away and HRP-labeled antihuman IgA or IgG antibody conjugate is added to each well. After a second incubation, unbound HRP-label is removed and bound conjugate is detected by adding TMB chromogenic substrate. After a final incubation, colored product is measured spectrophotometrically; the absorbance of the patient sample is compared to the positive calibrator. The absorbance is directly proportional to the level of IgA or IgG antibodies to deamidated gliadin peptides, which is expressed in arbitrary units.(Instruction manual: QUANTA Lite Gliadin IgA II and IgG II. INOVA Diagnostics, Inc. San Diego, CA, 92131)

 

Endomysial (EMA) Antibody, IgA:

IgA endomysial antibodies are detected by indirect immunofluorescence assay. Frozen sections of rhesus monkey esophagus substrate are overlaid with dilutions of patient sera and incubated. After washing, the slides are overlaid with fluorescein-conjugated IgA antisera and incubated. After a final washing, the slides are analyzed through fluorescence microscopy.(Chorzelski TP, Beutner EH, Sulet J, et al: IgA anti-endomysium antibody: A new immunological marker of dermatitis herpetiformis and coeliac disease. Br J Dermatol 1984;111:395-402)

 

PDF Report Indicates whether the report includes an additional document with charts, images or other enriched information

No

Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.

7 days

Performing Laboratory Location The location of the laboratory that performs the test

Rochester

Key