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Erschienen in: International Journal of Hematology 5/2019

04.03.2019 | Original Article

A novel method of amplified fluorescent in situ hybridization for detection of chromosomal microdeletions in B cell lymphoma

verfasst von: Yoshimi Mizuno, Yoshiaki Chinen, Taku Tsukamoto, Tomoko Takimoto-Shimomura, Yayoi Matsumura-Kimoto, Yuto Fujibayashi, Saeko Kuwahara-Ota, Takahiro Fujino, Daichi Nishiyama, Yuji Shimura, Tsutomu Kobayashi, Shigeo Horiike, Masafumi Taniwaki, Junya Kuroda

Erschienen in: International Journal of Hematology | Ausgabe 5/2019

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Abstract

Chromosomal microdeletions frequently cause loss of prognostically relevant tumor suppressor genes in hematologic malignancies; however, detection of minute deletions by conventional methods for chromosomal analysis, such as G-banding and fluorescence in situ hybridization (FISH), is difficult due to their low resolution. Here, we describe a new diagnostic modality that enables detection of chromosomal microdeletions, using CDKN2A gene deletion in B cell lymphomas (BCLs) as an example. In this method, which we refer to as amplified-FISH (AM-FISH), a 31-kb fluorescein isothiocyanate (FITC)-conjugated DNA probe encoding only CDKN2A was first hybridized with the chromosome, and then labeled with Alexa Fluor 488-conjugated anti-FITC secondary antibody to increase sensitivity. CDKN2A signals were equally identifiable by AM-FISH and conventional FISH in normal mononuclear blood cells. In contrast, when two BCL cell lines lacking CDKN2A were analyzed, CDKN2A signals were not detected by AM-FISH, whereas conventional FISH yielded false signals. Furthermore, AM-FISH detected CDKN2A deletions in two BCL patients with 9p21 microdeletions, which were not detected by conventional FISH. These results suggest that AM-FISH is a highly sensitive, specific, and simple method for diagnosis of chromosomal microdeletions.
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Metadaten
Titel
A novel method of amplified fluorescent in situ hybridization for detection of chromosomal microdeletions in B cell lymphoma
verfasst von
Yoshimi Mizuno
Yoshiaki Chinen
Taku Tsukamoto
Tomoko Takimoto-Shimomura
Yayoi Matsumura-Kimoto
Yuto Fujibayashi
Saeko Kuwahara-Ota
Takahiro Fujino
Daichi Nishiyama
Yuji Shimura
Tsutomu Kobayashi
Shigeo Horiike
Masafumi Taniwaki
Junya Kuroda
Publikationsdatum
04.03.2019
Verlag
Springer Japan
Erschienen in
International Journal of Hematology / Ausgabe 5/2019
Print ISSN: 0925-5710
Elektronische ISSN: 1865-3774
DOI
https://doi.org/10.1007/s12185-019-02617-x

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