摘要
目的探讨液态芯片技术在检测β地中海贫血基因突变常见类型的应用价值。方法采用液态芯片技术,使用Luminex100多功能悬浮点阵检测仪及Lumi2 nex Data Collector Version1.7数据收集软件检测并通过突变标本与正常标本的荧光强度比值(Mutan/Normal)分析100份正常人标本及68份PCR反向杂交技术确认的β地中海贫血基因突变标本。结果100份正常标本检测结果正常。68份β地贫基因突变检测结果为密码子CD41/42(TCTT)突变37例(54.41%),IVS2-654(C→T)突变杂合12例(17.65%),-28(A→G)突变10例(14.71%),-17(A→T)突变5例(7.35%),CD71/72(+A)2例(2.94%),-29(A→G)突变杂合1例(1.47%)。密码子71/72(+A)复合IVS2-654(C→T)突变杂合1例,-28(A→G)复合密码子CD41/42-TCTT突变2例,单位点突变β地贫病人为95.59%(65/68),多位点突变为4.41%(3/68)。7种突变杂合类型与常用方法PCR-RDB检测结果为100%符合率。结论液态芯片技术能准确区分β地贫基因突变杂合复合突变,可提供快速、敏感的临床β地贫基因突变检测技术平台。
Objective To investigate the application value of liquid microarrays technology in detection of common genetic mutation types of β-thalassemia. Methods By using Muhi-Analyte Profiling （LabMAP） system （Luminex 100^TM） and Lumi2 nex Data Collector Version 1.7, liquid microarrays technology were performed to detect gene mutation in 100 normal samples and 68 samples of β-thalassemia with mutation types confirmed by PCR reverse hybridization （PCR-RDB）, through analyzing fluorescent intensity ratio of mutation sample and normal sample. Results 100 normal samples were tested to be normal. The types of gene mutation in 68 β-thalassemia cases were CD41/42 （TCTF）, IVS2-654 （C→T）, -28 （A→G）, -17 （A→T）, CD71/72 （＋A）, -29 （A→G）, with the frequencies of 54.41% （37/68）, 17.65% （12/68）, 14.71% （10/68）, 7.35% （5/68）, 2.94% （2/68）, 1.47% （1/68）, respectively. Among them, 1 case was 71/72 （＋A） combined with IVS2-654 （C→T） and 2 cases was-28 （A→G） combined with CD41/42 （TCTF）. The frequencies of single point mutation and multi-point mutation were 95.59% （65/ 68） and 4.41% （3/68）, respectively. The coincidence rate of 8 hybrid mutation types obtained from liquid microarrays and PCR-RDB was 100%. Conclusion The liquid microarrays are able to differentiate accurately between the heterozygous and compound mutation in β-thalassemia. This technology provides us a rapid and sensitive detection platform for clinical diagnosis of β-thalassemia mutation.
出处
《热带医学杂志》
CAS
2007年第7期642-644,共3页
Journal of Tropical Medicine
关键词
液态芯片技术
Β地中海贫血
基因分析
liquid microarrays technology
β-thalassemia
gene typing analysis
作者简介
张松（1953-），本科，副主任技师，主要从事血液学检验工作。
