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1.
Chem Commun (Camb) ; 48(75): 9412-4, 2012 Sep 28.
Article in English | MEDLINE | ID: mdl-22889872

ABSTRACT

We have combined the benefits of a TaqMan assay with surface enhanced Raman scattering (SERS), to generate a novel DNA detection method which provides increased sensitivity, with clear applications for disease identification through clinical testing. Target DNA detection limits by SERS were shown to be lower than conventional fluorescence detection and clinically relevant samples of methicillin-resistant Staphylococcus aureus were detected with high specificity.


Subject(s)
DNA/analysis , Polymerase Chain Reaction , Spectrum Analysis, Raman , Taq Polymerase/metabolism , DNA, Bacterial/analysis , Limit of Detection , Methicillin-Resistant Staphylococcus aureus/genetics , Surface Properties
2.
Analyst ; 134(1): 170-5, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19082189

ABSTRACT

Bone marrow-derived immune cells (macrophages) treated with gold and silver nanoparticles before fixation and dye staining have been analysed by multiple wavelength line scanning surface enhanced resonance Raman scattering (SERRS) mapping. The method yields high selectivity and sensitivity within short analysis times, identifying nanoparticle aggregates in secondary lysosomes. Using routine cell stains, the output from fluorescence, Raman and SERRS is quantified at four wavelengths of excitation, demonstrating the potential at longer biologically compatible wavelengths of using nanoparticles with cell stains for superior cell mapping.


Subject(s)
Macrophages/ultrastructure , Coloring Agents , Gold , Nanoparticles , Sensitivity and Specificity , Silver , Spectrum Analysis, Raman/methods
3.
Anal Chem ; 79(7): 2844-9, 2007 Apr 01.
Article in English | MEDLINE | ID: mdl-17326610

ABSTRACT

There is a continued interest in the development of new on-chip protocols for the determination of the causes of infectious disease. In this paper, we demonstrate the use of surface-enhanced resonance Raman scattering (SERRS) for detecting the clinically relevant nucleic acid sequences of Chlamydia trachomatis in a bead-based lab-on-a-chip format, incorporating a solid-phase sample clean-up on-chip. The assay uses streptavidinated polymer microspheres to capture a biotinylated PCR product of the oligonucleotide sequence, which was subsequently hybridized against a complementary rhodamine-labeled, Raman active probe. Central to the assay is an in-channel integrated microfilter, which was used to retain the microspheres, enabling the bound target to be separated from the rest of the sample as part of a solid-phase clean-up (thereby removing any contributions from the background). After washing, the bound Rhodamine labeled detection probe was released thermally from the microspheres by heating and was subsequently mixed on-chip with a stream of silver nanoparticles. The signal was detected downstream using a Raman spectrometer to collect the SERRS response. The assay offers several advantages over traditional laboratory methods, including: the speed of the assay on-chip, the potential for sample clean-up; and the low volume of sample required.


Subject(s)
Chlamydia trachomatis/chemistry , DNA/analysis , Nanoparticles/chemistry , Oligonucleotide Array Sequence Analysis/methods , Spectrum Analysis, Raman/methods , Streptavidin/chemistry , Base Sequence , Equipment Design , Polymerase Chain Reaction/methods , Sensitivity and Specificity , Silver/chemistry , Surface Properties
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