A quantitative immuno-PCR assay for the detection of mumps-specific IgG

J Immunol Methods. 2002 Dec 1;270(1):135-41. doi: 10.1016/s0022-1759(02)00325-3.

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.

MeSH terms

  • Antibodies, Viral / blood*
  • Antibodies, Viral / genetics
  • Antibodies, Viral / immunology
  • Humans
  • Immunoglobulin G / blood*
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Mumps / blood
  • Mumps / virology*
  • Mumps virus / genetics
  • Mumps virus / immunology
  • Mumps virus / isolation & purification
  • Nucleocapsid Proteins / genetics
  • Nucleocapsid Proteins / immunology*
  • Nucleoproteins / genetics
  • Nucleoproteins / immunology*
  • Polymerase Chain Reaction / methods*

Substances

  • Antibodies, Viral
  • Immunoglobulin G
  • Nucleocapsid Proteins
  • Nucleoproteins