title: [Application of multiplex nested RT- PCR assay for screening the fusion genes in acute myeloid leukemia and its clinical significance].
abstract: OBJECTIVE
  To investigate the clinical value of multiplex nested reverse transcription PCR (RT-nPCR) in screening acute myeloid leukemia(AML)fusion genes.
  
  
  METHODS
  A novel multiplex RT-nPCR assay was developed to detect 16 AML-related fusion genes (AML1-EVI1, AML1-ETO, AML1-MDS1, AML1-MTG16, MLL-AF9, MLL-AF6, MLL-AF10, MLL-ENL, MLL-MLL, PML-RARα, PLZFRARα, NPM1-RARα, CBFB-MYH11, DEK-CAN, SET-CAN and TLS-ERG) according to 2008 WHO classification of AML. The chromosome reciprocal translocations of 356 AML cases were detected by multiplex RT-nPCR and karyotyping. The positive samples were further confirmed by split- out PCR and FISH.
  
  
  RESULTS
  The fusion genes were detected in 172 patients with the positive detection rate of 48.31%(172/356), which was higher than that of karyotyping (31.46%) (χ²=70.314, P<0.01). Multiplex RT-nPCR is superior to karyotyping and FISH in identifying the rare, cryptic chromosome translocation (χ²=96.074, P<0.01).
  
  
  CONCLUSIONS
  The multiplex RT-nPCR used in this study can quickly, effectively and accurately screen the fusion genes in AML patients, which can provide important evidence for assessing diagnosis and treatment, and also provide necessary information for minimal residual disease (MRD) and prognosis.
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Junzhong
authors.initials: J
authors.lastname: Sun
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Ding
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Yihan
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Lyu
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Liu
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
authors.forename: Li
authors.initials: L
authors.lastname: Yu
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D000090243:Nucleophosmin; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D055815:Young Adult
chemical_list: C000716967:NPM1 protein, human; D015514:Oncogene Proteins, Fusion; D000090243:Nucleophosmin