title: Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia.
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.
  
  
  CONCLUSIONS
  This simple and cost-effective technique is suitable for monitoring CML patients during targeted therapy (tyrosine kinase inhibitors) especially in rural hospitals.
authors.affiliation: Faculty of Medical Technology, Mahidol University, Bangkok, Thailand.
authors.forename: Yanaphat
authors.initials: Y
authors.lastname: Manthawornsiri
authors.affiliation: National Nanotechnology Center, National Science and Technology Development Agency, Thailand Science Park, Pathum Thani, Thailand.
authors.forename: Duangporn
authors.initials: D
authors.lastname: Polpanich
authors.affiliation: Faculty of Medical Technology, Mahidol University, Bangkok, Thailand.
authors.forename: Vichanan
authors.initials: V
authors.lastname: Yamkamon
authors.affiliation: National Nanotechnology Center, National Science and Technology Development Agency, Thailand Science Park, Pathum Thani, Thailand.
authors.forename: Raweewan
authors.initials: R
authors.lastname: Thiramanas
authors.affiliation: Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Suradej
authors.initials: S
authors.lastname: Hongeng
authors.affiliation: Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Budsaba
authors.initials: B
authors.lastname: Rerkamnuaychoke
authors.affiliation: Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Saengsuree
authors.initials: S
authors.lastname: Jootar
authors.affiliation: Department of Chemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.
authors.forename: Pramuan
authors.initials: P
authors.lastname: Tangboriboonrat
authors.affiliation: Faculty of Medical Technology, Mahidol University, Bangkok, Thailand. kulachart.jan@mahidol.ac.th.
authors.forename: Kulachart
authors.initials: K
authors.lastname: Jangpatarapongsa
mesh_terms: D000328:Adult; D000818:Animals; D045744:Cell Line, Tumor; D057075:Enzyme Assays / Q000379:methods*; D005260:Female; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics* / Q000378:metabolism; D058185:Magnetite Nanoparticles; D008297:Male; D051379:Mice; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D012737:Sex Factors; D055815:Young Adult
chemical_list: D058185:Magnetite Nanoparticles; D012333:RNA, Messenger; D016044:Fusion Proteins, bcr-abl
keywords: BCR/ABL fusion gene; RQ-PCR; chronic myelogenous leukemia; colorimetric analysis; magnetic nanoparticles