2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences

Abstract

A new approach is presented for the indirect hybridocytochemical localization of specific nucleic acid sequences in microscopic preparations. The method is based on the application of probes modified with N-acetoxy-2-acetylaminofluorene. After hybridization, the 2-acetylaminofluorene-labelled probes are recognized by antibodies directed against modified guanosine and visualized immunocytochemically. This procedure has been optimized on two model objects: mouse satellite DNA in interphase nuclei and chromosomes, and kinetoplast DNA in Crithidia fasciculata. A first application that may be of clinical importance is given by the detection of human cytomegalovirus in infected human lung fibroblasts. Other potentials of this procedure are discussed. Its advantages are: (1) the simple, rapid and reproducible labelling procedure; (2) the high stability of both label and modified probes; (3) the feasibility of labelling both double-stranded (ds) and single-stranded (ss) probes (DNA as well as RNA); (4) the rapid and sensitive detection of hybrids.