Immuno-PCR: Very sensitive antigen detection by means of specific antibody-DNA conjugates
- Univ. of California, Berkeley (United States)
An antigen detection system, termed immuno-polymerase chain reaction (immuno-PCR), was developed in which a specific DNA molecule is used as the marker. A streptavidin-protein A chimera that possesses tight and specific binding affinity both for biotin and immunoglobulin G was used to attach a biotinylated DNA specifically to antigen-monoclonal antibody complexes that had been immobilized on microtiter plate wells. Then, a segment of the attached DNA was amplified by PCR. Analysis of the PCR products by agarose gel electrophoresis after staining with ethidium bromide allowed as few as 580 antigen molecules to be readily and reproducibly detected. Direct comparison with enzyme-linked immunosorbent assay with the use of a chimera-alkaline phosphatase conjugate demonstrates that enhancement in detection sensitivity was obtained with the use of immuno-PCR. Given the enormous amplification capability and specificity of PCR, this immuno-PCR technology has a sensitivity greater than any existing antigen detection system and, in principle, could be applied to the detection of single antigen molecules.
- OSTI ID:
- 7224378
- Journal Information:
- Science (Washington, D.C.); (United States), Vol. 258:5079; ISSN 0036-8075
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANTIGENS
BIOLOGICAL MARKERS
POLYMERASES
BIOCHEMICAL REACTION KINETICS
ANTIGEN-ANTIBODY REACTIONS
DNA
ELECTROPHORESIS
ENZYME IMMUNOASSAY
MONOCLONAL ANTIBODIES
SENSITIVITY ANALYSIS
ANTIBODIES
BIOASSAY
ENZYMES
IMMUNOASSAY
KINETICS
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
REACTION KINETICS
TRANSFERASES
550200* - Biochemistry