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.
Seyda
M
M
Pathology Institute, University of Cologne, FRG.
Scheele
T
T
Neumann
R
R
Krueger
G R
GR
D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis*; D001345:Autoradiography; D003587:Cytomegalovirus / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D006678:HIV / Q000235:genetics; D006566:Herpesviridae Infections / Q000175:diagnosis*; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D009693:Nucleic Acid Hybridization; D011933:Reagent Kits, Diagnostic
D015342:DNA Probes; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic