Piezoelectric Plate Sensor for in situ Genetic Detection of Hepatitis B Virus in Serum without DNA Isolation and Amplification

Soylu, Mehmet Çağrı

doi:10.17918/etd-4314

Cited by 1 publication

(2 citation statements)

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“…The only difference is that the sensor must be blocked with a higher concentration of bovine serum albumin (BSA) after probe immobilization and prior to detection to prevent non-specific binding as plasma and sera contain higher concentrations of serum albumin. 60 Finally, the current hybridization temperature is not high enough to denature dsDNA. Detection of dsDNA will require denaturing the dsDNA first prior to hybridization.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, the probe immobilized on the MPS surface was quantified using a quartz crystal microbalance (QCM) to be about 3-4 probes per 100 nm 2 . 60 Thus, the SH of the MPS was proven to be effective to facilitate the immobilization of the probe DNA. A schematic of LEM vibrations and that of WEM vibrations of a PEPS is shown in insets (I) and (II) of Fig.…”

Section: Peps Fabrication and Electrical Insulationmentioning

confidence: 99%

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Amplification-free in situ KRAS point mutation detection at 60 copies per mL in urine in a background of 1000-fold wild type

Kirimli

Shih

2016

Analyst

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We have examined in situ detection of single-nucleotide KRAS mutation in urine using a (Pb(Mg1/3Nb2/3)O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) coated with a 17-nucleotide (nt) locked nucleic acid (LNA) probe DNA complementary to the KRAS mutation. To enhance in situ mutant (MT) DNA detection specificity against the wild type (WT), the detection was carried out in a flow with a flow rate of 4 mL/min and at 63°C with the PEPS vertically situated at the center of the flow in which both the temperature and the flow impingement force discriminated the wild type. Under such conditions, PEPS was shown to specifically detect KRAS MT in situ with 60 copies/mL analytical sensitivity in a background of clinically-relevant 1000-fold more WT in 30 min without DNA isolation, amplification, or labeling. For validation, the detection was followed with detection in a mixture of blue MT fluorescent reporter microspheres (FRMs) (MT FRMs) that bound to only the captured MT and orange WT FRMs that bound to only the captured WT. Microscopic examinations showed that the captured blue MT FRMs still outnumbered the orange WT FRMs by a factor of 4 to 1 even though WT was 1000-fold of MT in urine. Finally, multiplexed specific mutation detection was demonstrated using a 6-PEPS array each with a probe DNA targeting one of the 6 codon-12 KRAS mutations.

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Section: Discussionmentioning

confidence: 99%

Section: Peps Fabrication and Electrical Insulationmentioning

confidence: 99%