Erschienen in:
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
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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.