EPSTEIN-BARR-VIRUS (EBV) ENCODED SMALL RNAS - TARGETS FOR DETECTION BY INSITU HYBRIDIZATION WITH OLIGONUCLEOTIDE PROBES
EPSTEIN-BARR-VIRUS (EBV) ENCODED SMALL RNAS - TARGETS FOR DETECTION BY INSITU HYBRIDIZATION WITH OLIGONUCLEOTIDE PROBES
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DOI：
10.1136/jcp.45.7.616
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发表时间：
1992-07-01
影响因子：
3.4
通讯作者：
SLAVIN, G
中科院分区：
文献类型：
--
作者：
KHAN, G;COATES, PJ;SLAVIN, G
目的：建立一种快速、灵敏、特异的非同位素原位杂交（NISH）检测福尔马林固定、石蜡包埋组织中eb病毒的方法。方法：利用EBER-1和EBER-2两种低分子量RNA (Epstein-Barr encoded RNA)，使潜伏感染EBV的细胞分泌大量eber。该方法使用地高辛标记的反义寡核苷酸，对应于EBER-1和EBER-2的序列。结果：使用这些探针，结合高温微波变性，确保该技术比其他原位杂交技术检测EBV的灵敏度高得多。此外，杂交信号在形态学上是不同的，只有细胞核而不是核仁发出阳性信号。未观察到与其他嗜淋巴性疱疹病毒感染的细胞交叉杂交。结论：该技术的敏感性、简易性和快速性使其成为诊断和研究该病毒在肿瘤疾病中的作用的理想方法。
Aims: 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.Conclusion: 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.