Genosensör ile yeni bir girişimsel olmayan doğum öncesi tanı yöntemi

Year 2021, Volume: 46 Issue: 1, 208 - 215, 31.03.2021

Umut Kökbaş , Levent Kayrın , Abdullah Tuli

Abstract

Amaç: Bu çalışmanın amacı, maternal kandaki “cell-free” fetal DNA'dan paternal mutasyon tayini için nanopolimer bazlı genosensör ile yeni bir prosedür geliştirmektir.
Gereç ve Yöntem: Kuvars-kristal mikrobalans (QCM) ve biyosensör teknolojilerini kullanılarak β-talasemi IVS1-110 mutasyonun analizine uygulanmıştır. Bu amaçla, mutasyona ait problar genosensör elektrodu üzerine PoliHema-MAC nanopolimeri aracılığıyla bağlandı ve daha sonra örnekler tespit amacıyla çalışma hücresine eklendi. Genosensörün optimizasyon, karakterizasyon, duyarlılık ve özgüllük çalışmaları yapıldı.
Bulgular: Biyosensör ölçümleri -0,6 - 0,8 V sabit akım aralığında döngüsel volatmetriyle elde edildi. Girişimsel olmayan doğum öncesi tanı testi yapılan fetüslerde ARMS yöntemiyle ortak sonuçlar elde edilmiştir. ARMS sonuçları genosensör sonuçları ile istatistiksel olarak karşılaştırıldığında çizilen ROC eğrisinde %99 uyumluluk oranı elde edilmiştir.
Sonuç: Geliştirilen biyosensör, yüzeyinde bulunan proba özgül olarak, fetüsün paternal mutasyonunun varlığını ortaya koyabilmektedir. Rutinde kullanılan geleneksel yöntemlerle karşılaştırıldığında, daha düşük maliyetli, hızlı sonuç verebilen, özgüllüğü ve saptama etkinliği yüksek olarak bulunmuştur.

Keywords

Genosensör, “Cell-Free” Fetal DNA, Nanopolimer

Supporting Institution

Çukurova Üniversitesi Bilimsel Araştırma Projeleri Birimi

Project Number

TDK-2017-8841

Thanks

Çukurova Üniversitesi Bilimsel Araştırma Projeleri Birimi'ne çalışmamızı desteklediği için teşekkür ederiz.

A novel non-invasive prenatal test technique with genosensor

Abstract

Purpose: The aim of this study is to develop a new procedure with nanopolymer based genosensor for the determination of paternal mutation from cell-free fetal DNA in maternal blood.
Materials and Methods: β-thalassemia IVS1-110 mutation analysis was performed using quartz-crystal microbalance (QCM) and biosensor technologies. For this purpose, the probes of the mutation were immobilized on the genosensor electrode surface by means of Poly Hema-MAC nanopolymer and then samples were added to the working cell for detection. Optimization, characterization, sensitivity and specificity studies of the genosensor were performed.
Results: Biosensor measurements were obtained between -0.6 and 0.8 V in a constant current range by cyclic voltammetry. Common results were obtained with ARMS method in fetuses who had non-invasive prenatal diagnosis tests. When the ARMS results were compared with the genosensor results statistically, a 99% concordance rate was obtained in the ROC curve drawn.
Conclusions: The developed biosensor can detect the presence of paternal mutation of the fetüs specific to the probe on its surface. Compared with the traditional methods used in routine, it was found to be more cost-effective, faster, with high specificity and detection efficiency.

Keywords

Genosensor, Cell-FreeFetal DNA, Nanopolymer

Project Number

TDK-2017-8841

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