title: Quantitative detection of circulating nucleophosmin mutations DNA in the plasma of patients with acute myeloid leukemia.
abstract: OBJECTIVE
  The aim of this study was to quantify the copies of circulating nucleophosmin (NPM) mutations DNA in the plasma of patients with acute myeloid leukemia (AML) and to explore the association of circulating NPM mutation levels with clinical characteristics.
  
  
  METHODS
  The presence of NPM mutations in 100 Chinese patients newly diagnosed with AML were identified by RT-PCR and sequencing analysis. Copies of circulating NPM mutation A (NPM mut.A) DNA in the plasma of mutation-positive cases were quantified by real-time quantitative PCR (qRT-PCR). Furthermore, the association of circulating NPM mutation levels and clinical characteristics was analyzed.
  
  
  RESULTS
  NPM mutations were identified in 37 of the 100 patients and all cases were NPM mut.A. The circulating NPM mut.A levels ranged from 0.35×10(8) copies/ml to 6.0×10(8) copies/ml in the 37 mutation-positive cases. The medium and quartile M (P25, P75) of the circulating NPM mut.A levels in patients classified as M2, M4 and M5 morphological subtypes were 1.35×10(8) (0.76×10(8), 1.91×10(8)) copies/ml, 1.81×10(8) (1.47×10(8), 2.2×10(8)) copies/ml and 2.50×10(8) (2.42×10(8), 3.05×10(8)) copies/ml, respectively. Circulating NPM mut.A levels were significantly higher in patients with the M5 subtype of AML compared to patients with the M2 and M4 subtypes (p=0.000, p=0.046). In addition, circulating NPM mut.A copies were significantly associated with a higher white blood cell count, platelet count and bone marrow blast percentage (p<0.05).
  
  
  CONCLUSIONS
  Our results suggest that circulating NPM mutations DNA assay serves as a complementary to the routine investigative protocol of NPM-mutated leukemia.
authors.affiliation: 1. Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Yixueyuan Road, Chongqing 400016, P.R.China.
authors.forename: Jing
authors.initials: J
authors.lastname: Quan
authors.affiliation: 2. Department of Laboratory Medicine, Yantai Yuhuangding Hospital, Yantai 264000, P.R.China.
authors.forename: Yu-jie
authors.initials: YJ
authors.lastname: Gao
authors.affiliation: 3. Center for Hematology, Southwest Hospital, Third Military Medical University, Chongqing 400038, P.R.China.
authors.forename: Zai-lin
authors.initials: ZL
authors.lastname: Yang
authors.affiliation: 4. Department of Laboratory Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R.China.
authors.forename: Hui
authors.initials: H
authors.lastname: Chen
authors.affiliation: 1. Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Yixueyuan Road, Chongqing 400016, P.R.China.
authors.forename: Jing-rong
authors.initials: JR
authors.lastname: Xian
authors.affiliation: 1. Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Yixueyuan Road, Chongqing 400016, P.R.China.
authors.forename: Shuai-shuai
authors.initials: SS
authors.lastname: Zhang
authors.affiliation: 1. Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Yixueyuan Road, Chongqing 400016, P.R.China.
authors.forename: Qin
authors.initials: Q
authors.lastname: Zou
authors.affiliation: 1. Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Yixueyuan Road, Chongqing 400016, P.R.China.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
mesh_terms: D000293:Adolescent; D000328:Adult; D004247:DNA / Q000097:blood; D004252:DNA Mutational Analysis / Q000379:methods; D005260:Female; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000097:blood / Q000235:genetics* / Q000473:pathology; D007958:Leukocyte Count; D008297:Male; D008875:Middle Aged; D009154:Mutation; D009687:Nuclear Proteins / Q000097:blood / Q000235:genetics*; D000090243:Nucleophosmin; D010976:Platelet Count; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D055815:Young Adult
chemical_list: C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D004247:DNA
keywords: acute myeloid leukemia; circulating DNA; mutation; nucleophosmin; real-time quantitative polymerase chain reaction.