Nanoresonator chip-based RNA sensor strategy for detection of circulating tumor cells: response using PCA3 as a prostate cancer marker

Sioss, James A.; Bhiladvala, Rustom; Pan, Weihua; Li, Mingwei; Patrick, Susan; Xin, Ping; Dean, Stacey L.; Keating, Christine D.; Mayer, Theresa S.; Clawson, Gary A.

doi:10.1016/j.nano.2011.11.009

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Nanoresonator chip-based RNA sensor strategy for detection of circulating tumor cells: response using PCA3 as a prostate cancer marker

James A. Sioss, PhD
- Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, USA
Rustom Bhiladvala, PhD
- Department of Electrical Engineering, Pennsylvania State University, University Park, Pennsylvania, USA
- Materials Research Institute, Pennsylvania State University, University Park, Pennsylvania, USA
- Current address for R. Bhiladvala: Department of Mechanical Engineering, University of Victoria, Victoria BC, Canada.
Weihua Pan, MS
- Department of Pathology, Biochemistry and Molecular Biology, and Gittlen Cancer Research Foundation, Hershey Medical Center, Hershey, Pennsylvania, USA
Mingwei Li, PhD
- Department of Electrical Engineering, Pennsylvania State University, University Park, Pennsylvania, USA
- Materials Research Institute, Pennsylvania State University, University Park, Pennsylvania, USA
- Current address for M. Li: Agilent Technologies, Santa Rosa, California.
Susan Patrick, BS
- Department of Pathology, Biochemistry and Molecular Biology, and Gittlen Cancer Research Foundation, Hershey Medical Center, Hershey, Pennsylvania, USA
Ping Xin, BS
- Department of Pathology, Biochemistry and Molecular Biology, and Gittlen Cancer Research Foundation, Hershey Medical Center, Hershey, Pennsylvania, USA
Stacey L. Dean, PhD
- Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, USA
Christine D. Keating, PhD
- Department of Chemistry, Pennsylvania State University, University Park, Pennsylvania, USA
Theresa S. Mayer, PhD
- Department of Electrical Engineering, Pennsylvania State University, University Park, Pennsylvania, USA
- Materials Research Institute, Pennsylvania State University, University Park, Pennsylvania, USA
Gary A. Clawson, PhD, MD
- Materials Research Institute, Pennsylvania State University, University Park, Pennsylvania, USA
- Department of Pathology, Biochemistry and Molecular Biology, and Gittlen Cancer Research Foundation, Hershey Medical Center, Hershey, Pennsylvania, USA
- Corresponding author: Hershey Medical Center, H059, Pennsylvania State University, 500 University Drive, Hershey, PA 17033, USA.

Received 6 June 2011; accepted 10 November 2011. published online 23 November 2011.
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Abstract

There is widespread interest in circulating tumor cells (CTCs) in blood. Direct detection of CTCs (often < 1/mL) is complicated by a number of factors, but the presence of ∼10³ to 10⁴ copies of target RNA per CTC, coupled with simple enrichments, can greatly increase detection capability. In this study we used resonance frequency shifts induced by mass-amplifying gold nanoparticles to detect a hybridization sandwich bound to functionalized nanowires. We selected PCA3 RNA as a marker for prostate cancer, optimized antisense binding sites, and defined conditions allowing single nucleotide mismatch discrimination, and used a hybrid resonator integration scheme, which combines elements of top-down fabrication with strengths of bottom-up fabrication, with a view to enable multiplexed sensing. Bound mass calculated from frequency shifts matched mass estimated by counting gold nanoparticles. This represents the first demonstration of use of such nanoresonators, which show promise of both excellent specificity and quantitative sensitivity.

Graphical Abstract

Sandwich hybridization assay for DD3 RNA. A primary ASO (green) is covalently attached to the NW surface. Incubation with DD3 RNA (blue) results in hybridization. Subsequent hybridization with the ASO:AuNP (red) completes the sandwich. Formation of the sandwich hybridization complex adds mass to the anchored NW nanoresonator, which results in a shift in the resonance frequency of the NW, which is quantitatively proportional to the number of complexes bound. The resonance frequency shift is measured using laser interferometry.