摘要
目的探讨SYBR-GreenⅠ实时荧光PCR(SYBR-PCR)结合融解曲线分析(D.C)技术进行α-珠蛋白生成障碍性贫血(α-地贫)基因诊断的价值。方法对22个家系中的37份样品采用4管SYBR-PCR结合D.C进行--SEA、-α3.7,-α4.2及非缺失型(αα)4个等位基因型的检测,进而作出基因诊断。同时运用单管多重PCR(mPCR)结合凝胶电泳技术进行基因检测加以对照确认。结果22个家系中6个家系检出有--SEA或-α3.7携带者,其中东南亚型缺失携带者(--SEA/αα)7例,右侧缺失携带者(-α3.7/αα)3例,东南亚型缺失与右侧缺失双重杂合子(--SEA/-α3.7)1例。2例为胎儿产前诊断。其中1例为Bart′s水肿胎儿(--SEA/--SEA),另1例为东南亚型缺失携带者(--SEA/αα)。用mPCR凝胶电泳技术得出的基因诊断结果与上述结果一致。结论SYBR-PCR结合D.C技术在α-地贫的基因诊断、携带者检出及产前诊断方面具有实用价值。
Objective To explore the value of real-time polymerase chain reaction（PCR） with SYBR-GreenⅠ（SYBR-PCR） combined with dissociation curve（D.C） analysis used to molecular diagnose of α-thalassemia.Methods Thirty-seven samples came from 22 families were diagnosed by using SYBR-PCR combined with D.C analysis.The——^SEA,-α^3.7,-α^4.2 and non-deletion-type（αα） 4 genotypes were detected.At the same time,the single tube multiplex PCR methods（mPCR）was used to confirm it.Results Six of 22 families were found to carry——^SEAor-α^3.7 genes.Seven cases were——^SEA /αα,3 samples were-α^3.7/αα,1 sample was——^SEA/-α^3.7.Two fetuses were performed prenatal diagnosis,1 fetus was Bart′s hydrous,another was——SEA/αα.The detection results of the technique were the same as that of mPCR.Conclusion The SYBR-PCR combined with D.C analysis can be used to the carrier detection and prenatal diagnosis of α-thalassemia.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2008年第15期1181-1182,共2页
Journal of Applied Clinical Pediatrics
