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Usefulness of long-distance inverse polymerase chain reaction for molecular detection of 14q32 translocation in a clinical setting.

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Title: Usefulness of long-distance inverse polymerase chain reaction for molecular detection of 14q32 translocation in a clinical setting.
Authors: Ishizaki(Uemura), Akiko / Sugahara, Kazuyuki / Tsuruda, Kazuto / Hasegawa, Hiroo / Yanagihara, Katsunori / Tsukasaki, Kunihiro / Yamada, Yasuaki / Kamihira, Shimeru
Issue Date: Nov-2008
Publisher: Informa Healthcare
Citation: Scandinavian Journal of Clinical and Laboratory Investigation, 68(7), pp.519-525; 2008
Abstract: All mature B-cell leukaemias and lymphomas have a clonal Ig gene recombination, and half of them have a reciprocal chromosomal translocation involving the 14q32 locus. The 14q32 translocation partners are variable, such as BCL-2, BCL-1 and BCL-6, thus accounting for the difficulty in molecular detection by the current genomic polymerase chain reaction (PCR) method. To identify B-cell clones efficiently with an Ig gene rearrangement and reciprocal inter-chromosomal translocation, we verified the usefulness, in a practical laboratory setting, of our modified long-distance inverse (LDI) PCR method for detecting IgH gene rearrangements involving inter- and intra-chromosomal segments. The total run time of this LDI PCR method was 5.5 h. Using 24 samples of mature B-cell leukaemias and lymphomas, the modified LDI PCR gave clonally rearranged amplicons in 83 % (20/24) of cases. Direct sequencing results of the amplicons revealed inter-chromosomal translocations in 5 cases (25 %) and intra-chromosomal rearrangements in the remaining 15 cases (75 %). The partners of the inter-chromosomal translocation consisted of the 11q13.3 segment containing a partial BCL1 sequence in 3 cases; 18q21.3 segment containing a partial BCL2 sequence in one case; and a segment of 7q11.2 in one case. We present an LDI PCR-based methodology for the efficient identification of 14q32 translocations, with modifications to reduce the total run time to within one day.
Keywords: B-cell clonality / IgH gene / lymphoma / PCR
URI: http://hdl.handle.net/10069/22530
ISSN: 00365513
DOI: 10.1080/00365510701858240
PubMed ID: 18609098
Rights: Copyright (c) 2008 Informa plc / This is an electronic version of an article published in Scandinavian Journal of Clinical and Laboratory Investigation, 68(7), 519-525: 2008 November. Scandinavian Journal of Clinical and Laboratory Investigation is available online at: http://www.informaworld.com/openurl?genre=article&issn=00365513&volume=68&issue=7&spage=519.
Type: Journal Article
Text Version: author
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/22530

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