Detection and differentiation of Epstein-Barr virus strains by in situ polymerase chain reaction.
An in situ polymerase chain reaction (IS-PCR) technique was used to detect and differentiate strains of episomal Epstein-Barr virus (EBV) in infected cells. IS-PCR was performed on cell monolayers in eight-chamber glass slides using EBV type-specific primer pairs conserved within the EBV-encoded nuclear antigen (EBNA) 3C region. The amplicons in the cells were detected by in situ hybridization using EBV type-1 and type-2 specific 5'-biotinylated oligonucleotide probes and avidin-conjugated alkaline phosphatase as secondary reagent. This method was successfully used to identify EBV strains not only in Burkitt's lymphoma cell lines but also in B cells obtained from a patient with infectious mononucleosis. The technique described on this report is a reliable method to detect latently infected EBV-positive cells and can potentially be used to identify and type EBV strains present in clinical specimens.
Tamaki
H
H
Department of Pediatrics, University of Massachusetts Medical School, Worcester, USA.
Byron
K S
KS
Sullivan
J L
JL
Somasundaran
M
M
D001402:B-Lymphocytes / Q000821:virology; D002460:Cell Line; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D019309:Epstein-Barr Virus Nuclear Antigens / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D013045:Species Specificity
D017931:DNA Primers; D004279:DNA, Viral; D019309:Epstein-Barr Virus Nuclear Antigens