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Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia
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Metadata
Document Title
Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia
Author
Manthawornsiri Y., Polpanich D., Yamkamon V., Thiramanas R., Hongeng S., Rerkamnuaychoke B., Jootar S., Tangboriboonrat P., Jangpatarapongsa K.
Name from Authors Collection
Affiliations
Faculty of Medical Technology, Mahidol University, Bangkok, Thailand; National Nanotechnology Center, National Science and Technology Development Agency, Thailand Science Park, Pathum Thani, Thailand; Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand; Department of Chemistry, Faculty of Science, Mahidol University, Bangkok, Thailand
Type
Article
Source Title
Journal of Clinical Laboratory Analysis
ISSN
08878013
Year
2016
Volume
30
Issue
5
Page
534-542
Open Access
Green
Publisher
John Wiley and Sons Inc.
DOI
10.1002/jcla.21899
Abstract
Background: Magnetic nanoparticles (MNPs) have been widely used in medical diagnostic research. In this work, two technologies, MNPs and polymerase chain reaction (PCR), were combined to increase detection sensitivity and specificity. A novel technique based on the MNPs-PCR enzyme-linked gene assay (MELGA) was developed for detection of the BCR/ABL abnormal gene in chronic myelogenous leukemia (CML) patients. Methods: An MNPs-labeled BCR forward primer and a biotin-labeled ABL reverse primer were used to specifically amplify the target gene. After magnetic separation, the PCR product bound to MNPs labeled with streptavidin-conjugated horseradish peroxidase was incubated with the peroxidase substrate and hydrogen peroxide to generate the colorimetric signal. Results: When compared with real-time quantitative-PCR (RQ-PCR), the MELGA technique exhibited an increased sensitivity of <1 fg with high specificity for the BCR/ABL fusion gene in CML patients. In addition, MELGA colorimetric results correlated well with the number of copies obtained from RQ-PCR. Conclusion: This simple and cost-effective technique is suitable for monitoring CML patients during targeted therapy (tyrosine kinase inhibitors) especially in rural hospitals. © 2015 Wiley Periodicals, Inc.
Funding Sponsor
Office of the Higher Education Commission; Mahidol University; National Science and Technology Development Agency; Thailand Research Fund; National Research Council of Thailand; Thailand Graduate Institute of Science and Technology
License
CC BY, a CC BY-NC or a CC BY-NC-ND
Rights
Wiley, China
Publication Source
Scopus