Bead-based dna diagnostic assay for chlamydia using nanoparticle-mediated surface-enhanced resonance raman scattering detection within a lab-on-a-chip format

TY - JOUR

T1 - Bead-based dna diagnostic assay for chlamydia using nanoparticle-mediated surface-enhanced resonance raman scattering detection within a lab-on-a-chip format

AU - Monaghan, P.B.

AU - McCarney, K.M.

AU - Ricketts, Alastair

AU - Littleford, R.E.

AU - Docherty, F.T.

AU - Smith, W.E.

AU - Graham, D.

AU - Cooper, J.

PY - 2007/4/1

Y1 - 2007/4/1

N2 - 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.

AB - 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.

KW - micromachined filter-chamber

KW - solid-phase extraction

KW - total analysis systems

KW - capillary-electrophoresis

KW - microfluidic device

KW - electrochromatography

KW - integration

KW - silver

KW - SERRS

KW - beds

KW - surface-enhanced resonance raman scattering

U2 - 10.1021/ac061769i

DO - 10.1021/ac061769i

M3 - Article

VL - 79

SP - 2844

EP - 2849

JO - Analytical Chemistry

T2 - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 7

ER -