Epstein Barr virus (EBV) encoded small RNAs: targets for detection by in situ hybridisation with oligonucleotide probes.
OBJECTIVE
To develop a rapid, sensitive, and specific non-isotopic in situ hybridisation (NISH) procedure for the detection of Epstein-Barr virus in formalin fixed, paraffin wax embedded tissues.


METHODS
Two low molecular weight RNAs, designated EBER-1 and EBER-2 (Epstein-Barr encoded RNA), were used: cells latently infected with EBV secrete large amounts of EBERs. The method uses digoxigenin labelled anti-sense oligonucleotides, corresponding to sequences in EBER-1 and EBER-2.


RESULTS
The use of these probes, in conjunction with high temperature microwave denaturation, ensured that the technique was considerably more sensitive than other in situ hybridisation techniques for detecting EBV. Furthermore, the hybridisation signal was morphologically distinct in that only the nucleus and not the nucleolus give a positive signal. No cross-hybridisation was observed with cells infected with other lymphotropic herpes viruses.


CONCLUSIONS
The sensitivity, simplicity, and rapidity of this technique make it ideal for diagnostic use, and for studies investigating the role of this virus in neoplastic disease.
Khan
G
G
Department of Histopathology, Bartholomew's Hospital, West Smithfield, London.
Coates
P J
PJ
Kangro
H O
HO
Slavin
G
G
D002460:Cell Line; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006689:Hodgkin Disease / Q000382:microbiology; D006801:Humans; D008970:Molecular Weight; D009303:Nasopharyngeal Neoplasms / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity
D015345:Oligonucleotide Probes; D012367:RNA, Viral