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

• Published in: Journal of immunological methods Vol. 270; no. 1; pp. 135 - 141
• Main Authors: McKie, Anne, Samuel, Dhanraj, Cohen, Bernard, Saunders, Nicholas A
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-12-2002; Elsevier
• Subjects: Antibodies, Viral - blood; Antibodies, Viral - genetics; Antibodies, Viral - immunology; Antibody detection; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Health. Pharmaceutical industry; Humans; Immuno-PCR; Immunoglobulin G - blood; Immunoglobulin G - genetics; Immunoglobulin G - immunology; Immunological technics applied to diagnosis; In vitro gene amplification. Pcr technique; Industrial applications and implications. Economical aspects; Methods. Procedures. Technologies; Mumps; 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; Real-time PCR; PBS; Antibody detection; iPCR; Immuno-PCR; BSA; SD; MMR; Real-time PCR; TaqMan; rNP; tDNA; PBST; LC; Mumps; ELISA; Human; IgG; Patient; Nucleoprotein; Serology; Immunity; Immunological method; Antigen; Infection; Polymerase chain reaction; Viral disease; Serum; Recombinant protein; Detection; Quantitative analysis
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(02)00325-3

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.