Comparative evaluation of non-radioactive in situ hybridization techniques for pathologic diagnosis of viral infection.

We have compared the efficiency of biotinylated DNA probes and various visualization techniques with 35S-labeled DNA probes in routine paraffin sections from the pathology service; both autopsy and biopsy tissue were investigated. Probes included DNA genomic fragments from cytomegalovirus (CMV), Epstein-Barr-Virus (EBV) and human immunodeficiency virus (HIV). Methods of detection by Pathogene II kit (ENZO) and Blue Gene kit (BRL) with visualization by AEC, NBT/BCIP, Immunogold (Janssen) and autoradiography were used. The study shows most satisfactory results by applying the Blue Gene-NBT/BCIP combination followed by the Immunogold technique. Data obtained by these techniques compare well to those of using radioactive DNA probes and autoradiography.