Synthesis of PCR-derived, digoxigenin-labeled DNA probes for in situ detection of Epstein-Barr early RNAs in Epstein-Barr virus-infected cells.
A PCR-derived digoxigenin-labeled DNA probe was used for for Epstein-Barr early RNA (EBER) in situ hybridization in formalin-fixed paraffin-embedded tissues. The results showed that the hybridization signal was morphologically distinct and the intensity of signal was comparable with those by RNA riboprobe. The advantages of using PCR-derived DNA probes for EBER in situ hybridization include: (1) the synthesis of digoxigenin-labeled DNA probes is easy and simple by PCR; (2) the labeled amplification product can be used as a probe without further purification; (3) DNA probes are potentially more stable than RNA probes; and (4) the preparation of DNA probes is relatively efficient and rapid. It is concluded that this technique is an ideal candidate for detection of EBER expression in clinical specimens.
Tsai
S T
ST
Department of Otolaryngology, National Cheng Kung University Medical College, Taipan, Taiwan.
Jin
Y T
YT
Wu
T C
TC
D001483:Base Sequence; D015342:DNA Probes / Q000138:chemical synthesis*; D004279:DNA, Viral / Q000032:analysis; D004076:Digoxigenin / Q000737:chemistry; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000821:virology; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis
D015342:DNA Probes; D004279:DNA, Viral; D012367:RNA, Viral; D004076:Digoxigenin