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学科主题: 临床医学
题名:
A rapid, highly accurate method for quantifying CALR mutant allele burden in persons with myeloproliferative neoplasms
作者: Yao, Qiu-Mei1; Zhou, Jiao1; Gale, Robert Peter2; Li, Jin-Lan1; Li, Ling-Di1; Li, Ning1; Chen, Shan-Shan1; Ruan, Guo-Rui1
关键词: Calreticulin ; Mutation ; Allele burden ; Myeloproliferative neoplasm ; Polymerase chain reaction ; Fragment length analysis
刊名: HEMATOLOGY
发表日期: 2015-10-01
DOI: 10.1179/1607845415Y.0000000009
卷: 20, 期:9, 页:517-522
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Hematology
研究领域[WOS]: Hematology
关键词[WOS]: JAK2 V617F MUTATION ; ESSENTIAL THROMBOCYTHEMIA ; CALRETICULIN MUTATIONS ; PRIMARY MYELOFIBROSIS ; POLYCYTHEMIA-VERA ; SUBTYPES
英文摘要:

Background: Calreticulin (CALR) mutations were recently identified in a substantial proportion of persons with essential thrombocythemia (ET) and with primary myelofibrosis (PMF) without JAK2(V617F). Consequently rapid, sensitive, and specific methods to detect and quantify these mutations are needed.

Methods: We studied samples from 1088 persons with myeloproliferative neoplasms (MPNs) including 421 JAK2V617F negative subjects with ET, PMF, polycythemia vera (PV), chronic myeloid leukemia (CML) and hyper-eosinophilic syndrome (HES). Detection of CALR exon 9 mutations was done by PCR amplification followed by fragment length analysis and direct sequencing. Dilution assays were used to determine CALR mutant allele burden.

Results: We detected CALR mutations in blood and bone marrow samples from 152 subjects with ET and with PMF but not in samples from normal or persons with PV, CML, or HES. CALR mutant peaks were distinct from wild-type peaks and dilution experiments indicated a sensitivity level of 0.5-5% for a CALR mutant allele in a wild-type background. Diverse types of mutations were detected including deletions, insertions, and complex indels. All mutations were confirmed by direct sequencing. We also used dilution experiments to quantify mutant allele burden. We were able to reproducibly detect mutant allele levels as low 5% (0.5-5%) in a wild-type background.

Conclusions: PCR amplification followed by fragment length analysis is a rapid, sensitive, and specific method for screening persons with MPNs for CALR mutations, especially those with ET and PMF and for estimating mutant allele burden.

语种: 英语
所属项目编号: 2013CB733701 ; 81230013 ; 7122199 ; 20130001110079 ; 81170484
项目资助者: National Basic Research Program of China ; Key Program of National Natural Science Foundation of China ; Beijing Municipal Natural Science Foundation ; Specialized Research Fund for the Doctoral Program of Higher Education of China ; National Natural Science Foundation of China
WOS记录号: WOS:000362601900005
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/58073
Appears in Collections:北京大学第二临床医学院_血液科_期刊论文

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作者单位: 1.Peking Univ, Peoples Hosp, Inst Hematol, Beijing 100044, Peoples R China
2.Univ London Imperial Coll Sci Technol & Med, Dept Med, Div Expt Med, Hematol Res Ctr, London W12 OHS, England

Recommended Citation:
Yao, Qiu-Mei,Zhou, Jiao,Gale, Robert Peter,et al. A rapid, highly accurate method for quantifying CALR mutant allele burden in persons with myeloproliferative neoplasms[J]. HEMATOLOGY,2015,20(9):517-522.
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