Method Description

Immunoglobulin M:

The presence or absence of IgM-class antibody to varicella-zoster virus (VZV) is determined by an indirect immunofluorescence assay. Serum is incubated with VZV antigen that is adhered to a glass microscope slide. Antibodies, if present, will bind to the antigen forming stable antigen-antibody complexes. If no antibodies are present, the complexes will not be formed, and the serum components will be washed away. Fluorescein-labeled antihuman-IgM antibody is added to the reaction side and binds to IgM antibodies if present. This results in a positive reaction of bright apple-green fluorescence when viewed with a fluorescence microscope.(Package insert: Bion Varicella Zoster Antigen Substrate Slide. Bion Enterprises; 09/2019)

Immunoglobulin G:

The BioPlex 2200 VZV IgG assay uses multiplex flow immunoassay technology. Briefly, serum samples are mixed and incubated at 37° C with sample diluent and dyed beads coated with VZV antigen. After a wash cycle, antihuman IgG antibody conjugated to phycoerythrin (PE) is added to the mixture and incubated at 37° C. Excess conjugate is removed in another wash cycle, and the beads are resuspended in wash buffer. The bead mixture then passes through a detector that identifies the bead based on dye fluorescence and determines the amount of antibody captured by the antigen based on the fluorescence of the attached PE. Raw data is calculated in relative fluorescence intensity.

Three additional dyed beads, an internal standard bead, a serum verification bead, and a reagent blank bead are present in each reaction mixture to verify detector response, the addition of serum to the reaction vessel and the absence of significant nonspecific binding in serum.(Package insert: BioPlex 2200 System MMRV IgG. Bio-Rad Laboratories; 02/2019)