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Unit Code 88680:
West Nile Virus (WNV) Antibody, IgG and IgM, Spinal Fluid

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Method Description

IgG:

Polystyrene microwells are coated with recombinant WNV antigen.

Diluted specimens and controls are incubated in the wells to

allow specific antibody present in the specimens to react with the

antigen. Nonspecific reactants are removed by washing, and

peroxidase-conjugated antihuman IgG is added and reacts with

specific IgG. Excess conjugate is removed by washing. Enzyme

substrate and chromogen are added, and the color is allowed to

develop. After adding the Stop Reagent, the resultant color change

is quantified by a spectrophotometric reading of optical density

(OD). Specimen OD readings are compared with reference cut-off

readings to determine results. (Package insert: Flavivirus [West Nile]

ELISA IgG. Focus Technologies, Cypress, CA)

 

IgM:

Polystyrene microwells are coated with the antihuman antibody

specific for IgM (u-chain). Diluted specimens and controls are

incubated in the wells, and IgM present in the specimen binds to the

antihuman antibody (IgM specific) in the wells. Nonspecific reactants

are removed by washing. WNV antigen is then added to the wells

and incubated. If  anti-WNV IgM is present in the specimen, the WNV

antigen binds to the anti-WNV in the well. Unbound WNV antigen is

then removed by washing the well. Mouse antiflavivirus conjugated

with horseradish peroxidase (HRPO) is then added to the wells and

incubated. If WNV antigen has been retained in the well by the

antiflavivirus in the specimen, the mouse antiflavivirus:HRPO binds

to WNV antigen in the wells. Excess conjugate is removed by washing.

Enzyme substrate and chromogen are added, and the color is allowed

to develop. After adding the Stop Reagent, the resultant color change

is quantified by a spectrophotometric reading of OD that is directly

proportional to the amount of antigen-specific IgM present in the

specimen. Specimen OD readings are compared with reference

cutoff OD readings to determine results. (Package insert: Flavivirus

[West Nile] IgM Capture ELISA. Focus Technologies, Cypress CA)

Performing Laboratory Location

Rochester

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