用于β-39地中海贫血突变实时分子诊断的表面等离子体共振和生物传感器技术
Surface plasmon resonance and biosensor technology for real-time molecular diagnosis of beta o 39 thalassemia mutation.
作者信息
Feriotto Giordana, Breveglieri Giulia, Gardenghi Sara, Carandina Gianni, Gambari Roberto
机构信息
Laboratory for the Development of Pharmacological and Pharmacogenomic Treatment of Thalassemia, Biotechnology Center, Ferrara University, Ferrara, Italy.
出版信息
Mol Diagn. 2004;8(1):33-41. doi: 10.1007/BF03260045.
BACKGROUND
Biospecific interaction analysis (BIA) employing surface plasmon resonance (SPR) and biosensor technologies is of interest in clinical genetics. However, few data are available on its use in hereditary diseases caused by genetic mutations.
AIM
The primary aim of this study was the refinement of BIA technology for use in identifying the beta o 39 mutation of the beta-globin gene, a mutation which causes a common type of beta o thalassemia.
METHODS
Target-biotinylated PCR products were immobilized on streptavidin-coated sensor chips and diagnosed using SPR-based BIA performed by injecting specific oligonucleotide probes into the sensor chip.
RESULTS
We demonstrated that the beta o 39 mutation can be easily and reproducibly identified during the association phase.
CONCLUSIONS
This should be considered a pilot study demonstrating the ability of SPR-based BIA to detect point mutations in the beta-globin gene by real-time monitoring of hybridization between oligonucleotide probes and target-biotinylated PCR products generated from genomic DNA from normal, heterozygous individuals and homozygous beta o thalassemia patients.
背景
采用表面等离子体共振（SPR）和生物传感器技术的生物特异性相互作用分析（BIA）在临床遗传学领域备受关注。然而，关于其在由基因突变引起的遗传性疾病中的应用数据较少。
目的
本研究的主要目的是优化BIA技术，用于鉴定β-珠蛋白基因的βo 39突变，该突变导致一种常见类型的βo地中海贫血。
方法
将靶向生物素化的PCR产物固定在链霉亲和素包被的传感器芯片上，并通过向传感器芯片中注入特异性寡核苷酸探针，采用基于SPR的BIA进行诊断。
结果
我们证明在结合阶段可以轻松且可重复地鉴定出βo 39突变。
结论
这应被视为一项初步研究，证明了基于SPR的BIA能够通过实时监测寡核苷酸探针与从正常个体、杂合子个体和纯合βo地中海贫血患者的基因组DNA产生的靶向生物素化PCR产物之间的杂交，来检测β-珠蛋白基因中的点突变。
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