NONISOTOPIC LABELING OF DNA BY NEWLY DEVELOPED HAPTEN-CONTAINING PLATINUM COMPOUNDS

A newly developed reagent was tested for its applicability in in situ hybridization and in reversed hybridization of DNA fragments generated by PCR amplification. This Platinum-complex, designated universal lin kage system (ULS), equipped, for instance, with biotin or fluorescein as hapten, enables versatile nonenzymatic ''one step'' labeling of gen omic, cloned or amplified DNA. Here we demonstrate direct in situ dete ction of integrated human papilloma virus (HPV) DNA in cervical carcin oma cells using DNA probes labeled with fluorescein-ULS. In cervical s mears the presence of HPV or Chlamydia trachomatis was assessed by PCR . To analyze the amplified DNA, a reversed hybridization assay was dev eloped. Immobilized probes were incubated with amplimers that were lab eled post-amplification through the action of the biotinylated (BIO)-U LS complex. This novel type of nonradioactive analysis appeared to be as sensitive as its isotopic or colorimetric equivalents. This labelin g procedure is simple, versatile and can be included as a universal ha pten linkage system in any PCR test or in situ hybridization assay aim ing at the detection and identification of DNA or RNA molecules.