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Specific In Situ Hepatitis B Viral Double Mutation (HBVDM) Detection in Urine with 60 copies/ml Analytical Sensitivity in a background of 250-fold Wild Type without DNA Isolation and Amplification
Journal article   Open access

Specific In Situ Hepatitis B Viral Double Mutation (HBVDM) Detection in Urine with 60 copies/ml Analytical Sensitivity in a background of 250-fold Wild Type without DNA Isolation and Amplification

Ceyhun E Kirimli, Wei-Heng Shih and Wan Y Shih
Analyst (London), v 140(5), pp 1590-1598
07 Mar 2015
PMID: 25599103
url
https://doi.org/10.1039/c4an01885kView
Published, Version of Record (VoR) Open

Abstract

We have examined in situ detection of hepatitis B virus 1762T/1764A double mutation (HBVDM) in urine using a (Pb(Mg 1/3 Nb 2/3 )O 3 ) 0.65 (PbTiO 3 ) 0.35 (PMN-PT) piezoelectric plate sensor (PEPS) coated with a 16-nucleotide (nt) probe DNA (pDNA) complementary to the HBVDM. The in situ mutation (MT) detection was carried out in a flow with the PEPS vertically situated at the center of the flow in a background of the wild type (WT). For validation, the detection was followed with detection in the mixture of MT fluorescent reporter microspheres (FRMs) (MT FRMs) and WT FRMs that emitted different fluorescent colours and were designed to specifically bind to MT and WT, respectively. At 30°C and 4 ml/min, a PEPS was shown to specifically detect HBVDM in situ with 60 copies/ml analytical sensitivity in a background of clinically-relevant 250-fold more WT in 30 min without DNA isolation, amplification, or labelling as validated by the visualization of the captured MT FRMs and WT FRMs following the FRMs detection where the captured MT FRMs outnumbered WT FRMs by a factor of 5 to 1.

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Web of Science research areas
Chemistry, Analytical
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