title: Standard polymerase chain reaction analysis does not detect t(14;18) in reactive lymphoid hyperplasia.
abstract: There are conflicting data regarding the detection of t(14;18) in reactive lymphoid hyperplasia (RLH) by the polymerase chain reaction (PCR). Although most studies have not detected t(14;18), several groups have definitively shown that a very low number of cells with this translocation (one in 10(5) to 10(6)) are present in a significant proportion of follicular hyperplasias. Review of the methods from these series reveals that modifications of the PCR assay (ie, enhanced sensitivity steps such as seminesting, lengthy autoradiographic exposure times, multiple aliquot reactions of single samples, and/or high concentrations of template DNA) are probably necessary to detect t(14;18) in RLH. We evaluated a diverse set of 111 RLH (85 lymph nodes, 22 tonsils, and four other sites) from patients of different age groups (age range, 9 months to 80 years) to determine if a standard PCR assay would amplify t(14;18). Of these, 61 (55%) specimens had a prominent follicular hyperplastic component. Fifty-seven follicular lymphomas served as a control group. Polymerase chain reaction was performed as a single-run, two-primer-based assay for major breakpoint region bcl-2 translocations (5' major breakpoint region primer and 3' immunoglobulin heavy-chain gene-joining region consensus primer). Two different types of thermocyclers were employed. A metal block thermocycler was used with 35 cycles of amplification on 500 ng to 1 micrograms of genomic DNA, and a separate air thermocycler was used with 45 cycles of amplification on 50 ng of genomic DNA. Product detection was carried out through ethidium bromide staining and UV gel illumination, along with a digoxigenin-alkaline phosphatase-based, internal major breakpoint region oligonucleotide probe system. We found no amplified t(14;18) products in any RLH. In contrast, 36 (63%) of 57 follicular lymphomas showed t(14;18) (published range for detection of major breakpoint region translocations by PCR, 31% to 74%). Moreover, the assay's sensitivity, estimated through dilution studies, was to one in 10(4) to 10(5) cells. Although theoretically possible, our data suggest that there is practically no risk of amplifying a t(14;18) from RLH when utilizing a standard PCR assay.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville 32610.
authors.forename: G H
authors.initials: GH
authors.lastname: Segal
authors.forename: M
authors.initials: M
authors.lastname: Scott
authors.forename: T
authors.initials: T
authors.lastname: Jorgensen
authors.forename: R C
authors.initials: RC
authors.lastname: Braylan
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D002883:Chromosomes, Human, Pair 14; D002887:Chromosomes, Human, Pair 18; D005260:Female; D006801:Humans; D006965:Hyperplasia; D007223:Infant; D008221:Lymphoid Tissue / Q000473:pathology*; D008224:Lymphoma, Follicular / Q000235:genetics; D008297:Male; D008875:Middle Aged; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014178:Translocation, Genetic
chemical_list: D015335:Molecular Probes