IMMERSION-HISTO POLYMERASE CHAIN-REACTION - A PRACTICAL TOOL FOR VISUALIZATION OF SINGLE-COPY GENES IN TISSUE-SECTIONS

A method, immersion-histo polymerase chain reaction (IH-PCR), was deve loped for visualization of single-copy DNA sequences in cells in paraf fin-embedded tissue sections. Sections were mounted on coverslips, cut into small pieces, and immersed in reaction mixtures in micro-tubes f or specific DNA amplification using a conventional thermal cycler, Thi s was followed by in situ hybridization, in micro-tubes, with PCR-gene rated, digoxigenein-labeled probes. Epstein-Barr virus, chromosomal tr anslocations, and rearranged immunoglobulin genes were clearly demonst rated in individual cells in human lymphomas. The use of a standard th ermal cycler, the ease of probe preparation, and the high specificity and sensitivity of target localization make IH-PCR a practical and pow erful tool for the study of genetic events within individual cells.