Method Description

IgM:

The SeraQuest mumps IgM assay is an enzyme capture method. Diluted samples are incubated in wells coated with antihuman-IgM monoclonal antibodies. If present, IgM antibodies are captured in the wells. Wells are washed, removing excess sample. Conjugate-antigen complex (mumps antigen in complex with monoclonal antibodies conjugated to horseradish peroxidase) is added, and the wells are incubated. IgM antibodies specific for the antigen will bind the conjugate. Wells are washed, removing excess conjugate. Peroxidase substrate is added, and the wells are incubated. Stop solution is added converting the substrate to a yellow end product, which is read photometrically.(Package insert: Mumps IgM. Quest International; V 04/2018)

IgG:

The BioPlex 2200 Mumps 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 mumps 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 by 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)