Primers Frequently Used for Detecting the tlpar;14semi;18rpar; Major Breakpoint also Amplify Epsteinhyphen;Barr Viral DNA

摘要

NA;We observed a potentially misinterpretable polymerase chain reaction lpar;PCRrpar; amplification product generated with standard primers used to detect the major breakpoint region lpar;mbrrpar; of chromosomal translocation tlpar;14semi;18rpar;. This unexpected phenomenon was initially detected during attempts to transform follicular lymphomas in vitro with Epsteinhyphen;Barr virus lpar;EBVrpar;. Additional studies were performed using the EBVhyphen;producing cell line MCUV5, cell lines from EBVhyphen;transformed normal Bhyphen;lymphocytes, and an excised lymph node from a patient with documented EBVhyphen;associated infectious mononucleosis. These samples consistently produced a 167hyphen;base pair product, which was indistinguishable from a tlpar;14semi;18rpar; lymphoma product when viewed on ethidium bromidehyphen;stained gels. Through DNA sequencing and gene bank analysis, the product was identified as a portion of the EBV genome. A mbrspecific 20hyphen;base oligonucleotide probe was able to discriminate between true translocations and the EBVhyphen;related amplifications. These results underscore the importance of employing a specific detection system, and comprehensively screening primers when working with PCR.