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Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation

Identification of fusion genes in cancer is essential for pathological diagnosis and clinical therapy. Although methods for detection of fusion genes, such as fluorescence in situ hybridization (FISH) and real-time polymerase chain reaction (PCR), have been developed in last two decades, these metho...

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Autores principales: Matsuzaki, Ibu, Iguchi, Hideto, Mikasa, Yurina, Morishita, Hiromu, Okuda, Katsuya, Nakaguchi, Keita, Mori, Yuki, Iwahashi, Yoshifumi, Warigaya, Kenji, Fujimoto, Masakazu, Kojima, Fumiyoshi, Murata, Shin-ichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5765217/
https://www.ncbi.nlm.nih.gov/pubmed/29343880
http://dx.doi.org/10.1267/ahc.17024
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author Matsuzaki, Ibu
Iguchi, Hideto
Mikasa, Yurina
Morishita, Hiromu
Okuda, Katsuya
Nakaguchi, Keita
Mori, Yuki
Iwahashi, Yoshifumi
Warigaya, Kenji
Fujimoto, Masakazu
Kojima, Fumiyoshi
Murata, Shin-ichi
author_facet Matsuzaki, Ibu
Iguchi, Hideto
Mikasa, Yurina
Morishita, Hiromu
Okuda, Katsuya
Nakaguchi, Keita
Mori, Yuki
Iwahashi, Yoshifumi
Warigaya, Kenji
Fujimoto, Masakazu
Kojima, Fumiyoshi
Murata, Shin-ichi
author_sort Matsuzaki, Ibu
collection PubMed
description Identification of fusion genes in cancer is essential for pathological diagnosis and clinical therapy. Although methods for detection of fusion genes, such as fluorescence in situ hybridization (FISH) and real-time polymerase chain reaction (PCR), have been developed in last two decades, these methods are not ideal for detection of these genetic alterations owing to their high cost and time-consuming procedures. In this study, we developed novel application for detection of gene translocations using loop-mediated isothermal amplification (LAMP). We verified the amplified DNA products of echinoderm microtubule-associated protein-like 4 and anaplastic lymphoma kinase (EML4-ALK), synaptotagmin and synovial sarcoma, X breakpoint (SYT-SSX), and immunoglobulin heavy chain gene and B cell leukemia/lymphoma 2 (IgH/BCL2) by real-time PCR, agarose-gel electrophoresis, and the naked eye after the LAMP procedure. Fusion genes were detected in samples diluted 10(3) times within 60 min. Because of the advantages of rapid amplification, simple operation, and easy detection without requiring sophisticated equipment or technical skill, LAMP may have potential applications as an on-site analytical approach in hospitals for pathological diagnosis and decision making regarding appropriate therapeutic approachs.
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spelling pubmed-57652172018-01-17 Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation Matsuzaki, Ibu Iguchi, Hideto Mikasa, Yurina Morishita, Hiromu Okuda, Katsuya Nakaguchi, Keita Mori, Yuki Iwahashi, Yoshifumi Warigaya, Kenji Fujimoto, Masakazu Kojima, Fumiyoshi Murata, Shin-ichi Acta Histochem Cytochem Regular Article Identification of fusion genes in cancer is essential for pathological diagnosis and clinical therapy. Although methods for detection of fusion genes, such as fluorescence in situ hybridization (FISH) and real-time polymerase chain reaction (PCR), have been developed in last two decades, these methods are not ideal for detection of these genetic alterations owing to their high cost and time-consuming procedures. In this study, we developed novel application for detection of gene translocations using loop-mediated isothermal amplification (LAMP). We verified the amplified DNA products of echinoderm microtubule-associated protein-like 4 and anaplastic lymphoma kinase (EML4-ALK), synaptotagmin and synovial sarcoma, X breakpoint (SYT-SSX), and immunoglobulin heavy chain gene and B cell leukemia/lymphoma 2 (IgH/BCL2) by real-time PCR, agarose-gel electrophoresis, and the naked eye after the LAMP procedure. Fusion genes were detected in samples diluted 10(3) times within 60 min. Because of the advantages of rapid amplification, simple operation, and easy detection without requiring sophisticated equipment or technical skill, LAMP may have potential applications as an on-site analytical approach in hospitals for pathological diagnosis and decision making regarding appropriate therapeutic approachs. JAPAN SOCIETY OF HISTOCHEMISTRY AND CYTOCHEMISTRY 2017-12-26 2017-12-07 /pmc/articles/PMC5765217/ /pubmed/29343880 http://dx.doi.org/10.1267/ahc.17024 Text en 2017 The Japan Society of Histochemistry and Cytochemistry This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Regular Article
Matsuzaki, Ibu
Iguchi, Hideto
Mikasa, Yurina
Morishita, Hiromu
Okuda, Katsuya
Nakaguchi, Keita
Mori, Yuki
Iwahashi, Yoshifumi
Warigaya, Kenji
Fujimoto, Masakazu
Kojima, Fumiyoshi
Murata, Shin-ichi
Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title_full Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title_fullStr Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title_full_unstemmed Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title_short Novel Application of Loop-mediated Isothermal Amplification for Rapid Detection of Gene Translocation
title_sort novel application of loop-mediated isothermal amplification for rapid detection of gene translocation
topic Regular Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5765217/
https://www.ncbi.nlm.nih.gov/pubmed/29343880
http://dx.doi.org/10.1267/ahc.17024
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