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	用探针熔解曲线法检测急性髓细胞白血病c-kit基因突变; Scanning of c-kit mutations in acute myeloid leukemia using probe melting curve method
	姜国强 ; 赵晓明 ; 王航 ; 李庆阁 ; 鹿全意
	2013-05-01
关键词	白血病 原癌基因蛋白质c-kit 突变 熔解曲线分析 Leukemia Proto-oncogene proteins c-kit Mutation Melting curve analysis
英文摘要	目的用改良的杂交探针结合溶解曲线新方法检测急性髓性白血病C-kIT基因17号外显子上基因突变。方法利用PrIMEr PrEMIErV5.00、TM uTIlITyV1.3等软件设计引物探针,两个阶段探针分别针对d816、n820和n822等突变热点。取患者dnA样品5μl进行PCr扩增及熔解曲线分析,PCr扩增产物进行测序分析并与熔解曲线分析方法进行比较。结果阳性质粒灵敏度试验的结果显示,n820g的灵敏度为10%,其余突变灵敏度均能达到5%。12例Cbf-AMl标本中,5例检测为阳性,突变率为41.7%,测序结果与测序检测结果相符。结论新方法具有方便快速、灵敏度高、特异性好等特点,可用于Cbf-AMl患者的个体化诊断和治疗。; Objective To detect c-kit gene mutations of acute myeloid leukemia in the hotspot exon 17 of the c-kit gene using a modified hybridization probe combining with melting curve.Methods Primer Premier v5.00 and Tm Utility v1.3 software were used to design the primers and probe,the probe contained two parts,the first segment of the probe could detect mutations around D816 and the second segment could detect mutations at N820 and N822.5 μl DNA samples from patients were used for PCR amplification and melting curve analysis.PCR products were sent to be sequenced,these results were compared with those of the mutation-positive plasmids.Results The detection sensitivity on positive control plasmids was 10% for N820G and 5% for the other mutations.c-kit mutations were identified in 5 of 12 patients with CBF-AML(41.7%),the result was confirmed by sequencing analysis.Conclusion The new method is simple,rapid,and specific with high sensitivity and could be used in individual diagnosis and treatment of CBF-AML patients.
语种	zh_CN
内容类型	期刊论文
源URL	[http://dspace.xmu.edu.cn/handle/2288/119215]
专题	生命科学－已发表论文
推荐引用方式 GB/T 7714	姜国强,赵晓明,王航,等. 用探针熔解曲线法检测急性髓细胞白血病c-kit基因突变, Scanning of c-kit mutations in acute myeloid leukemia using probe melting curve method[J],2013.
APA	姜国强,赵晓明,王航,李庆阁,&鹿全意.(2013).用探针熔解曲线法检测急性髓细胞白血病c-kit基因突变..
MLA	姜国强,et al."用探针熔解曲线法检测急性髓细胞白血病c-kit基因突变".(2013).

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