Y-chromosome identification in circulating cell-free fetal DNA using surface plasmon resonance

被引:16
作者
Breveglieri, Giulia [1 ,2 ]
Bassi, Elisabetta [1 ]
Carlassara, Silvia [1 ]
Cosenza, Lucia Carmela [1 ,2 ]
Pellegatti, Patrizia [3 ]
Guerra, Giovanni [3 ]
Finotti, Alessia [1 ]
Gambari, Roberto [1 ,2 ]
Borgatti, Monica [1 ]
机构
[1] Univ Ferrara, Biochem & Mol Biol Sect, Dept Life Sci & Biotechnol, I-44100 Ferrara, Italy
[2] Univ Ferrara, Biotechnol Ctr, I-44100 Ferrara, Italy
[3] Univ Hosp S Anna, Lab Anal, Operat Unit, Ferrara, Italy
关键词
REAL-TIME DETECTION; STAPHYLOCOCCAL-ENTEROTOXIN-B; FIBROSIS W1282X MUTATION; PEPTIDE NUCLEIC-ACIDS; MATERNAL PLASMA; BIOSENSOR TECHNOLOGY; SEX DETERMINATION; PRENATAL-DIAGNOSIS; PREGNANT-WOMEN; KINETICS;
D O I
10.1002/pd.4788
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
ObjectiveSince the discovery of cell-free fetal DNA (cffDNA) in maternal plasma, diagnostic non-invasive prenatal methods have been developed or optimized for fetal sex determination and identification of genetic diseases. As far as fetal sex determination, this might be important for therapeutic intervention on sex-associated pathologies such as Duchenne muscular dystrophy, hemophilia and congenital adrenal hyperplasia. Surface plasmon resonance (SPR)-based biosensors might be useful for these studies, because they allow to monitor the molecular interactions in real-time providing qualitative and quantitative information, through kinetics, affinity and concentration analyses. MethodsThe Biacore(TM) X100 has been applied to identify Y-chromosome sequence in cffDNA obtained from plasma samples of 26 pregnant women at different gestational ages. We have performed SPR-based analysis of SRY PCR products using SRY-specific probes immobilized on the sensor chip. ResultsWe have demonstrated that there is a statistically significant difference between samples collected by pregnancies carrying male or female fetuses. Moreover, cffDNA obtained at early gestational ages and not detectable by conventional quantitative real-time PCR can be discriminated with high accuracy and reliability using SPR-based biosensors. ConclusionsThese data, in addition to their direct applicability in more extensive diagnostic trials, should be considered as the basis of future developments. (c) 2016 John Wiley & Sons, Ltd.
引用
收藏
页码:353 / 361
页数:9
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