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Science 2 October 1992:
Vol. 258. no. 5079, pp. 120 - 122
DOI: 10.1126/science.1439758
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Articles

Science, Vol 258, Issue 5079, 120-122
Copyright © 1992 by American Association for the Advancement of Science

articles

Immuno-PCR: very sensitive antigen detection by means of specific antibody-DNA conjugates

T Sano, CL Smith, and CR Cantor

Department of Molecular and Cell Biology, University of California, Berkeley 94720.

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 (9.6 x 10(-22) moles) 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 (approximately x 10(5)) 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.


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