Primers frequently used for detecting the t(14;18) major breakpoint also amplify Epstein-Barr viral DNA.
We observed a potentially misinterpretable polymerase chain reaction (PCR) amplification product generated with standard primers used to detect the major breakpoint region (mbr) of chromosomal translocation t(14;18). This unexpected phenomenon was initially detected during attempts to transform follicular lymphomas in vitro with Epstein-Barr virus (EBV). Additional studies were performed using the EBV-producing cell line MCUV5, cell lines from EBV-transformed normal B-lymphocytes, and an excised lymph node from a patient with documented EBV-associated infectious mononucleosis. These samples consistently produced a 167-base pair product, which was indistinguishable from a t(14;18) lymphoma product when viewed on ethidium bromide-stained gels. Through DNA sequencing and gene bank analysis, the product was identified as a portion of the EBV genome. A mbr-specific 20-base oligonucleotide probe was able to discriminate between true translocations and the EBV-related amplifications. These results underscore the importance of employing a specific detection system, and comprehensively screening primers when working with PCR.
Segal
G H
GH
Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville.
Scott
M
M
Jorgensen
T
T
Braylan
R C
RC
D000818:Animals; D001483:Base Sequence; D002144:Callithrix; D002883:Chromosomes, Human, Pair 14; D002887:Chromosomes, Human, Pair 18; D017931:DNA Primers / Q000737:chemistry* / Q000592:standards; D004279:DNA, Viral / Q000737:chemistry*; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014178:Translocation, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004279:DNA, Viral