荧光PCR-探针熔解曲线法在缺失型α-地中海贫血快速诊断中的应用  

Application of fluorescence PCR-probe melting curve analysis in the rapid diagnosis of deletionα-thalassemia

作　　者：李恋湘 肖奇志[1] 张素粉 吴洪秋[1] LI Lian-xiang;XIAO Qi-zhi;ZHANG Su-fen;WU Hong-qiu(Department of Clinical Laboratory,Zhuhai Municipal Maternity and Child Healthcare Hospital,Guangdong Province,Zhuhai519001,China)

机构地区：[1]广东省珠海市妇幼保健院检验科,广东珠海519001

出　　处：《中国当代医药》2020年第15期24-27,F0004,共5页China Modern Medicine

摘　　要：目的探讨荧光PCR-探针熔解曲线法(PMCA)在缺失型α-地中海贫血(简称“α-地贫”)快速诊断中的应用。方法收集2019年10~12月珠海市妇幼保健院2261份血样本和35份羊水样本。通过双盲法,采用PMCA和单管多重PCR技术(gap-PCR)同时对上述2296份临床样本进行3种常见缺失型α-地贫基因检测,结果不符样本用DNA测序技术以确认。结果2296份临床样本中,除PMCA检测出1例未知突变基因之外,其余与gap-PCR检测结果一致,两者符合率为99.9%,此例经DNA测序显示在探针覆盖区域发生了单个碱基突变(A>G)。结论PMCA技术能快速准确检测3种常见缺失型α-地贫基因,可作为gap-PCR技术的替代或验证方法,也可用于大规模人群筛查及产前诊断。Objective To explore the application of fluorescence PCR-probe melting curve analysis(PMCA)in the rapid diagnosis of deletionα-thalassemia.Methods A total of 2261 blood samples and 35 amniotic fluid samples were collected in Zhuhai Municipal Maternity and Child Healthcare Hospital from October to December 2019.Through double-blind method,PMCA and single-tube multiplex PCR(gap-PCR)were used to detect three common deletionα-thalassemia genes in these 2296 clinical samples.The samples inconsistent with results were confirmed by DNA sequencing analysis.Results In these 2296 samples,except for one unknown mutation gene detected by PMCA in one sample,the results of PMCA in others were consistent with those of gap-PCR,with a coincidence rate of 99.9%.DNA sequencing of the only inconsistent sample showed that a single base mutation(A>G)occurred in the probe coverage area.Conclusion PMCA can quickly and accurately detect three common deletionα-thalassemia genes,which can be used as an alternative or verification method of gap-PCR technology,as well as for large-scale population screening and prenatal diagnosis.

关 键 词：探针 熔解曲线 Α-地中海贫血 多重聚合酶链反应 

分 类 号：R714.55[医药卫生—妇产科学]