Comparison of polymerase chain reaction and standard Southern blotting for the detection of Epstein-Barr virus DNA in various biopsy specimens.
The sensitivity of the polymerase chain reaction (PCR) assay was compared to that of standard Southern blotting (SB) hybridization for detecting the presence of Epstein-Barr virus (EBV) genomes in biopsy samples from 43 patients with a variety of lymphoproliferative disorders. Two pairs of oligonucleotide primers from the first BamHI M and R leftward reading frames (BMLF1 and BRLF1) of EBV were chosen to amplify DNA. The resulting PCR products were analyzed by gel electrophoresis, transfer and hybridization. Restriction enzyme digestion was used to confirm the specificity of the amplified fragment. EBV DNA was found in 38 of 43 patients, as compared with 9 of 43 patients with the Southern technique. No amplified product was detected with other viruses from the Herpes family, nor with human genomic DNA from healthy adults using the same two sets of primers. These results indicate that EBV can be detected in a greater number of lymphoproliferative lesions than previously appreciated. The implications of these findings are discussed.
Department of Medical Microbiology, Université de Montréal, Québec, Canada.
L
L
Pedneault
B Z
BZ
Katz
D000293:Adolescent; D000328:Adult; D001483:Base Sequence; D001706:Biopsy; D015139:Blotting, Southern; D002648:Child; D002675:Child, Preschool; D004262:DNA Restriction Enzymes; D004277:DNA, Single-Stranded; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics*; D015245:Deoxyribonuclease BamHI; D005260:Female; D005814:Genes, Viral; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D008223:Lymphoma / Q000175:diagnosis / Q000382:microbiology; D008232:Lymphoproliferative Disorders / Q000175:diagnosis / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012680:Sensitivity and Specificity
D004277:DNA, Single-Stranded; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D004262:DNA Restriction Enzymes; D015245:Deoxyribonuclease BamHI