A nonradioactive hybridization assay for the detection of hepatitis B virus (HBV) DNA in serum with a digoxigenin-labeled probe is described. The probe was sensitive, being able to detect 0.25 pg of homologous HBV DNA, equivalent to 7 x 10(4) genome copies. After extraction of DNA from clinical samples, the probe detected HBV DNA in 11 of 12 hepatitis B e antigen-positive sera and did not react...

We report microtiter well-based sandwich-type DNA hybridization assays using enzyme amplified time-resolved fluorometry of Tb3+ chelates. The target DNA was hybridized with two adjacent and non-overlapping oligonucleotide probes, one oligonucleotide serving as the capture probe and the other as the detection probe. Two ligand-specific binding protein pairs were used alternately for capture of t...

This study was undertaken to develop technology for the detection of nucleic acid using two different DNA probe reporter molecules, the ultimate aim being to differentially label two nucleic acids within the same nucleus. Digoxigenin and biotin were used to label DNA probes. The absolute and relative sensitivity of digoxigenin and biotin labelled DNA probes for detecting integrated human papill...

Summary DNA nanotechnology offers new biosensing approaches by templating different sensor and transducer components. Here, we combine origami nanoantennas with label-free antibody detection incorporating a nanoswitch in the plasmonic hotspot of nanoantenna. The contains two antigens that are displaced binding, thereby eliciting fluorescent signal. Single-antibody is demonstrated integ...

The development of biosensors for detection and identification of DNA sequences by hybridization may reduce assay time and allow direct quantitation of the target. This article describes the use of impedance spectroscopy technique for digoxigeninthiol-labeled ssDNA probe immobilization, anti-digoxigenin binding and targeted DNA hybridization. The analysis of the impedance spectra in terms of eq...

We have developed a system for the enzymatic in vitro synthesis of non-radioactively labeled RNA which is derivatized with the hapten digoxigenin (DIG). The labeling reaction as well as the conditions for hybridization and detection of hybrids by an antibody-conjugate and a coupled colour reaction were analyzed and adapted for high sensitivity and low background. In addition, data on the perfor...

The digoxigenin derivative N-hydroxysuccinimidyl digoxigenin-3-O-methylcarbonyl-epsilon-aminocaproate (HDMA) has been shown to covalently label the ouabain binding site of the Na,K-ATPase epsilon subunit [Antolovic et al. (1995) Eur. J. Biochem. 227, 61-67]. In the present study we observed both, labeling and inactivation of the activity, of wild type Na,K-ATPase overexpressed in Xenopus oocyte...

Figure 2A shows an example of a whole-mount embryo double-labeled with a fluorescein-labeled fushi tarazu (ftz) and a digoxigenin labeled wingless (wg) probe. Several other combinations of differentially labeled probes were tested including a fluorescein-labeled ftzprobe together with a digoxigenin-labeled engrailed(en) probe, a fluores-cein-labeled wgprobe together with a digoxigenin-labeled e...

Microfluidic chip was successfully fabricated from polydimethylsiloxane (PDMS) and used for direct detection of E.coli O157:H7 in ground beef via anti-digoxigenin immuno-PCR. The PDMS channels were functionalized with 100 μg/ml streptavidin at room temperature for 6 min. A recovery of 60% ± 3.4 was obtained when bacterial cells were extracted from seeded ground beef using differential centrifug...