title: A quantitative immuno-PCR assay for the detection of mumps-specific IgG.
abstract: Sensitive assays are required for seroprevalence studies of measles, mumps and rubella (MMR)-vaccinated populations where many may have low levels of antibodies. This protocol describes a quantitative immuno-PCR assay to detect mumps-specific IgG antibodies. The purpose of the protocol is to determine the immune status of individuals to mumps. Mumps-specific IgG from a dilution of patients serum is bound by recombinant mumps nucleoprotein coated on the surface of microtitre plate wells. Bound antibody is detected by PCR using a conjugate of anti-human IgG covalently coupled to an oligonucleotide. The oligonucleotide is detected by the addition of target DNA, designed to hybridise to the oligonucleotide and serve as a template for real-time PCR using the LightCycler. The quantity of target DNA detected by the PCR depends upon the level of specific antibody in the test sample.
authors.affiliation: Genomics, Proteomics and Bioinformatics Services, Central Public Health Laboratory, 61 Colindale Avenue, Colindale, London NW9 5HT, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: McKie
authors.forename: Dhanraj
authors.initials: D
authors.lastname: Samuel
authors.forename: Bernard
authors.initials: B
authors.lastname: Cohen
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Saunders
mesh_terms: D000914:Antibodies, Viral / Q000097:blood* / Q000235:genetics / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood* / Q000235:genetics / Q000276:immunology; D009107:Mumps / Q000097:blood / Q000821:virology*; D009109:Mumps virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins