Takeuchi H, Kobayashi R, Hasegawa M, Hirai K
Department of Cell Regulation, Tokyo Medical and Dental University, Japan.
J Virol Methods. 1996 Apr 26;58(1-2):81-9. doi: 10.1016/0166-0934(95)01993-6.
A highly sensitive and specific reverse transcription-polymerase chain reaction (RT-PCR) assay was designed for detection of Epstein-Barr virus (EBV)-related sequences in nucleic acid extracted by the conventional phenol method. Using this EBV-infected malignant lymphoma cell line, Raji cells, a comparative study of this assay was carried out with EBER1 (EBV-encoded small RNA1) primer and conventional DNA-PCR with BamHI-W and EBER1 primers, respectively. The results revealed that this assay has sensitivity about 10(5)-fold higher than the conventional DNA-PCR method. The presence of EBER1 DNA and RNA was also investigated in 23 healthy individuals and 22 right and left tonsils of 11 healthy individuals. These results indicated that this assay is both sensitive and specific. Thus, EBV infection could be diagnosed easily determined and EBER1 was shown to be transcribed in peripheral blood cells and tonsils with quantitatively different grades. This assay can be used to diagnose EBV infection in clinical samples.
设计了一种高灵敏度和特异性的逆转录-聚合酶链反应（RT-PCR）检测方法，用于检测通过传统苯酚法提取的核酸中与爱泼斯坦-巴尔病毒（EBV）相关的序列。使用这种EBV感染的恶性淋巴瘤细胞系Raji细胞，分别用EBER1（EBV编码的小RNA1）引物和用BamHI-W和EBER1引物的传统DNA-PCR对该检测方法进行了比较研究。结果显示，该检测方法的灵敏度比传统DNA-PCR方法高约10^5倍。还对23名健康个体以及11名健康个体的22个左右扁桃体进行了EBER1 DNA和RNA的检测。这些结果表明该检测方法既灵敏又特异。因此，可以轻松诊断EBV感染，并且显示EBER1在外周血细胞和扁桃体中以不同的定量水平转录。该检测方法可用于临床样本中EBV感染的诊断。
