A chemical method for introducing haptens onto DNA probes

• Published in: Analytical biochemistry Vol. 170; no. 2; p. 441
• Main Authors: Keller, G H, Cumming, C U, Huang, D P, Manak, M M, Ting, R
• Format: Journal Article
• Language: English
• Published: United States 01-05-1988
• Subjects: 2,4-Dinitrophenol; Affinity Labels; Animals; Cloning, Molecular; Dinitrophenols - analysis; Dinitrophenols - immunology; DNA - analysis; Haptens; Hepatitis B virus - genetics; Humans; Nucleic Acid Hybridization; Rabbits; Viral Proteins - analysis; Viral Proteins - genetics
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(88)90656-2

We have developed a versatile chemical method of attaching hapten moieties onto DNA, for the construction of nonisotopic DNA probes. The DNA is reacted with N-bromosuccinimide at alkaline pH, resulting in bromination of a fraction of the thymine, guanine, and cytosine residues, with adenine modified to a lesser extent. The bromine is subsequently displaced by a primary amino group, attached to a linker arm. The other end of the linker arm has a detectable group preattached to it. We have labeled cloned hepatitis B viral (HBV) DNA with the hapten 2,4-dinitrophenyl (DNP) and used it in combination with a high affinity rabbit anti-DNP antibody, for the detection of hepatitis B DNA by slot blotting. This probe was sensitive enough to specifically detect 1 X 10(-17) mol (1 X 10(6) copies) of HBV DNA in total DNA from human serum.