Detection of latent infection by Epstein-Barr virus in peripheral blood cells of healthy individuals and in non-neoplastic tonsillar tissue from patients by reverse transcription-polymerase chain reaction.
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.
Takeuchi
H
H
Department of Cell Regulation, Tokyo Medical and Dental University, Japan.
Kobayashi
R
R
Hasegawa
M
M
Hirai
K
K
D002051:Burkitt Lymphoma / Q000097:blood / Q000821:virology*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D015870:Gene Expression; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D014066:Palatine Tonsil / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D017735:Virus Latency
D017931:DNA Primers; D004279:DNA, Viral; D015245:Deoxyribonuclease BamHI