Detection of Epstein-Barr virus DNA in Hodgkin- and Reed-Sternberg-cells by single cell PCR.
The Epstein-Barr virus (EBV) can be detected in the majority of lymph nodes involved by Hodgkin's lymphoma using the highly sensitive polymerase chain reaction (PCR). However, the rate of EBV-DNA detection by in-situ hybridisation, which allows allocation of EBV to a defined cell population, i.e. the neoplastic H&RS-cells, is lower. In an attempt to combine the advantages of the high sensitivity of the PCR and the possibility of cellular allocation by in-situ hybridisation, we established a single-cell PCR of Hodgkin- and Reed-Sternberg (H&RS)-cells isolated by micromanipulation from biopsy tissues. We amplified EBV sequences from the BamW-region by single-cell PCR. Using this method we were able to detect EBV-DNA in the H&RS-cells from 4 of 6 patients. In EBV positive cases all H&RS-cells of a given patient were positive, proving the high sensitivity and reproducibility of the method. Other cells in the biopsy tissue involved by EBV-positive H&RS-cells were shown to be negative. This indicates that EBV may have a role in the pathogenesis of many but not all cases of Hodgkin's disease.
Innere Medizin I, Universitätskliniken des Saarlandes, Homburg, Germany.
J
J
Roth
H
H
Daus
A
A
Gause
L
L
Trümper
M
M
Pfreundschuh
D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D001706:Biopsy; D017931:DNA Primers; D005260:Female; D004854:Herpesvirus 4, Human / Q000302:isolation & purification*; D006689:Hodgkin Disease / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D008198:Lymph Nodes / Q000382:microbiology* / Q000473:pathology; D008297:Male; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction / Q000379:methods*; D016539:Reed-Sternberg Cells / Q000382:microbiology* / Q000473:pathology
D017931:DNA Primers; D016376:Oligonucleotides, Antisense