2010 NS-AUA Abstracts.
Abstract: Background:

Urinary tract infections (UTI) are among the most common bacterial infections in the United States. The gold standard for diagnosis is urine culture. This provides bacteria identification and sensitivity, but is labor intensive and time-consuming. As such, many UTIs are treated empirically contributing to resistance and cost. There is an urgent need for a diagnostic tool that can rapidly identify uropathogens and their sensitivities. We present the development of a PCR-based assay for Enterobacter cloacae, and evidence for its further use in the context of NanoLantern technology. The NanoLantern is a chip sensor that utilizes DNA hairpins immobilized onto a gold surface. In the presence of complementary (target) DNA, the hairpin stem is opened, exposing a quantifiable fluorophore.

Methods:

A synthetic DNA hairpin, unique to Enterobacter cloacae, was immobilized onto 9 gold coated chips. Baseline fluorescence intensity was recorded. The chip was exposed to synthetic target and then imaged with a fluorescence microscope. Fluorescence intensity was recorded and compared to baseline. PCR, asymmetric PCR and linear after the exponential PCR were used to amplify the target DNA from the bacteria. The amplicon was exposed to the NanoLantern chip and fluorescence intensity was quantified.

Results:

Pre-synthetic target fluorescence imaging of 9 chips exposed to synthetic DNA resulted in a mean raw count of 665 (SD +- 244). Post-synthetic target fluorescence imaging of 9 chips resulted in a mean raw count of 8197 (SD +- 648), a greater than 12-fold increase in intensity. See Fig. 1.

Conclusions:

We have demonstrated the feasibility of a novel DNA hairpin probe. Work is ongoing to develop a PCR assay and to begin testing the NanoLantern chip with bacteria derived amplicon and urine samples. Ultimately, these integrated assays will provide rapid analysis, as part of an automated system, to be used in the clinical setting.
Authors: Cambio, Angelo J.
Rothschild, Jennifer G.
Peng, Hsin-P.
Mayer, Robert D.
Miller, Benjamin L.
Pub Date: 10/01/2010
Publication: Name: Canadian Urological Association Journal (CUAJ) Publisher: Canadian Urological Association Audience: Academic Format: Magazine/Journal Subject: Health Copyright: COPYRIGHT 2010 Canadian Urological Association ISSN: 1911-6470
Issue: Date: Oct 1, 2010 Source Volume: 4 Source Issue: 5 Suppl 2
Accession Number: 239214078
Full Text: Author(s): Angelo J. Cambio [1], Jennifer G. Rothschild [1], Hsin-P Peng [2], Robert D. Mayer [1], Benjamin L. Miller [2]

We wish to acknowledge and thank Watson Pharmaceuticals, Inc. for their support in the publication of the Northeastern Secion - AUA 2010 Annual Meeting abstracts.

Author Affiliation(s):

[1] University of Rochester Medical Center, Rochester, NY, USA;

[2] University of Rochester, Rochester, NY, USA

[3] University of Pittsburgh Medical Center, Pittsburgh, PA, USA

[4] Universite Laval, Quebec, QC, Canada;

[5] University of California, Los Angeles, Los Angeles, CA, USA

[6] University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

[7] Queen's University, Kingston, ON, Canada

[8] LOEX, Centre de recherche FRSQ du CHA Universitaire de Quebec, Universite Laval, Quebec, QC, Canada

[9] Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada;

[10] St Joseph's Health Care London, London, ON, Canada

[11] CPSI Biotech, Owego, NY, USA;

[12] Binghamton University, Binghamton, NY, USA;

[13] Duke University Medical Center, Durham, NC, USA

[14] CPSI Biotech, Owego, NY, USA;

[15] Binghamton University, Binghamton, NY, USA

[16] University of Rochester, Rochester, NY, USA

[17] University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

[18] University of Pittsburgh Medical Center, Pittsburgh, PA, USA

[19] Queen's University, Kingston, ON, Canada

[20] Queen's University, Kingston, ON, Canada

[21] University of British Columbia, New Westminster, BC, Canada

[22] University of Pittsburgh Medical Center, Pittsburgh, PA, USA

[23] William Beaumont Hospital, Royal Oak, MI, USA

[24] CPSI Biotech, Owego, NY, USA;

[25] Binghamton University, Binghamotn, NY, USA;

[26] Duke University Medical Center, Durham, NC, USA;

[27] Binghamton University, Binghamton, NY, USA

[28] McGill University, Montreal, QC, Canada;

[29] University of Texas Southwestern Medical Center, Dallas, TX, USA;

[30] University of Padua, Padua, Italy;

[31] Kitasato University School of Medicine, Sagamihara, Japan;

[32] Derby City General Hospital, Derby, United Kingdom;

[33] Caritas St. Josef Medical Center, University of Regensburg, Regensburg, Germany;

[34] Ludwig-Maximilians-University, Klinikum Grosshadern, Munich, Germany;

[35] Hospital Universitario Puerta de Hierro-Majadahonda, Universidad Autónoma de Madrid, Madrid, Spain;

[36] General Hospital Bolzano, Bolzano, Italy;

[37] University of Montreal, Montreal, QC, Canada

[38] Roswell Park Cancer Institute, Buffalo, NY, USA;

[39] Cedars-Sinai Medical Center, Los Angeles, CA, USA

[40] University of Rochester, Rochester, NY, USA

[41] University of Toronto, Toronto, ON, Canada

[42] Princess Margaret Hospital, Toronto, ON, Canada

[43] Cancer Prognostics and Health Outcomes Unit, University of Montreal Health Centre, Montreal, QC, Canada;

[44] Prostate Cancer Center, University Medical Center Hamburg-Eppendorf, Hamburg, Germany;

[45] Urology, Centre Hospitalier Universitaire de Montpellier, Montpellier, France;

[46] Department of Urology, Vita-Salute San Raffaele University, Milan, Italy;

[47] Department of Urology, University of Montreal Health Center, Montreal, QC, Canada;

[48] Prostate Cancer Centre, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

[49] Cancer Prognosis and Health Outcomes Unit, University of Montreal, Montreal, QC, Canada;

[50] Department of Urology, Vita-Salute San Raffaele University, Milan, Italy;

[51] Department of Urology, Prostate Cancer Center Hamburg-Eppendorf,, Hamburg, Germany;

[52] Department of Urology, Montpellier University Hospital, Montpellier, France;

[53] Department of Urology, Prostate Cancer Center Hamburg-Eppendorf, Hamburg, Germany;

[54] Department of Urology, University of Montreal, Montreal, QC, Canada

[55] McGill University, Montreal, QC, Canada

[56] Roswell Park Cancer Institute, Buffalo, NY, USA

[57] University of Montreal Health Centre, Montreal, QC, Canada;

[58] Cancer Prognostics and Health Outcomes Unit, Montreal, QC, Canada;

[59] Vita-Salute University San Raffaele, Milan, Italy;

[60] Martiniclinic, University Hospital Hamburg-Eppendorf, Hamburg, Germany

[61] Department of Urology, Cancer Prognostics and Health Outcomes Unit, University of Montreal, Montreal, QC, Canada;

[62] Department of Urology, Vita-Salute San Raffaele University, Milan, Italy;

[63] Urology, Centre Hospitalier Universitaire de Montpellier, Montpellier, France;

[64] Prostate Cancer Centre, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

[65] University of Montreal Health Centre, Montreal, QC, Canada;

[66] Cancer Prognostics and Health Outcomes Unit, Montreal, QC, Canada;

[67] Martiniclinic, University Hospital Hamburg-Eppendorf, Hamburg, Germany

[68] Albany Medical College, Albany, NY, USA

[69] University of Rochester Medical Center, Rochester, NY, USA

[70] University of Rochester Medical Center, Rochester, NY, USA

[71] University of Toronto, Toronto, ON, Canada

[72] University of Rochester, Rochester, NY, USA

[73] University of Toronto, Toronto, ON, Canada

[74] McGill University, Montreal, QC, Canada

[75] University of Pittsburgh, Pittsburgh, PA, USA

[76] Schulich School of Medicine and Dentistry, The University of Western Ontario, London, ON, Canada

[77] University of Toronto, Toronto, ON, Canada

[78] McGill University, Montreal, QC, Canada

[79] McGill University, Montreal, QC, Canada

[80] University of Pittsburgh Medical Center, Pittsburgh, PA, USA;

[81] University of Pittsburgh Department of Pharmacology, Pittsburgh, PA, USA

[82] Clarion Hospital, Clarion, PA, USA;

[83] Lake Erie Osteopathic College of Medicine, Erie, PA, USA

[84] Butler Memorial Hospital, Butler, PA, USA

[85] University of Rochester, Rochester, NY, USA

[86] Universite Laval, Quebec, QC, Canada

[87] University of Western Ontario, London, ON, Canada

[88] University of Pittsburgh, Pittsburgh, PA, USA

[89] Virginia Mason Medical Center, Seattle, WA, USA

[90] University of Montreal Health Center, Montreal, QC, Canada;

[91] Martini Clinic Prostate Cancer Center, Hamburg-Eppendorf, Germany;

[92] Vita-Salute San Raffaele, Milan, Italy;

[93] Centre Hospitalier Universitaire de Montpellier, Montpellier, France;

[94] McGill University, Montreal, QC, Canada

[95] University of Montreal Health Center, Montreal, QC, Canada;

[96] Martini Clinic Prostate Cancer Center, Hamburg-Eppendorf, Germany;

[97] Vita-Salute San Raffaele, Milan, Italy;

[98] Centre Hospitalier Universitaire de Montpellier, Montpellier, France;

[99] McGill University, Montreal, QC, Canada

[100] Main Urology Associates P.C., Buffalo, NY, USA;

[101] Community Care, Albany, NY, USA;

[102] GTx, Inc., Memphis, TN, USA;

[103] GTx, Inc, Memphis, TN, USA

[104] University of Rochester, Rochester, NY, USA

[105] Grand Strand Urology, Myrtle Beach, SC, USA;

[106] Mount Sinai School of Medicine, New York, NY, USA;

[107] Eastern Virginia Medical School, Norfolk, VA, USA;

[108] University of Washington, Seattle, WA, USA;

[109] Virginia Mason Medical Center, Seattle, WA, USA;

[110] Oregon Health and Sciences University, Portland, OR, USA;

[111] University of California, San Francisco, San Francisco, CA, USA;

[112] University of California, Los Angeles, Los Angeles, CA, USA;

[113] Georgia Urology, Marietta, GA, USA;

[114] Dendreon Corporation, Seattle, WA, USA

[115] Queen's University, Kingston, ON, Canada

[116] McGill University, Montreal, QC, Canada

[117] Virginia Mason Medical Center, Seattle, WA, USA

[118] Princess Margaret Hospital, Toronto, ON, Canada

[119] University of Ottawa, Ottawa, ON, Canada

[120] University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

[121] Albany Medical Center, Albany, NY, USA;

[122] Ordway Research Institute, Albany, NY, USA

[123] Clarion Hospital, Clarion, PA, USA

[124] McGill University, Montreal, QC, Canada

[125] McGill University, Montreal, QC, Canada

[126] UPMC St. Margaret, Kittanning, PA, USA

[127] Department of Urology, State University of New York at Buffalo, Erie County Medical Center, Buffalo, NY, USA

[128] Weill Cornell Medical College, New York, NY, USA;

[129] UT Southwestern Medical Center, Dallas, TX, USA;

[130] Watson Laboratories, Salt Lake City, UT, USA

[131] Urology Institute of Pittsburgh, Pittsburgh, PA, USA

[132] Virginia Mason Medical Center, Seattle, WA, USA

[133] University of Ottawa, Ottawa, ON, Canada

[134] York Central Hospital, Richmond Hill, ON, Canada

[135] Urology, Cancer Prognostics and Health Outcomes Unit, University of M, Montreal, QC, Canada;

[136] Department of Urology, Vita-Salute San Raffaele University, Milan, Italy;

[137] Department of Urology, Martini-Clinic, Prostate Cancer Center, University Medical Center Hamburg-Eppendorf, Hamberg, Germany;

[138] Department of Urology, Centre Hospitalier Universitaire de Montpellier, Montpellier, France;

[139] Department of Urology, Cancer Prognostics and Health Outcomes Unit, University of Montreal, Montreal, QC, Canada;

[140] Department of Urology, Vita-Salute San Raffaele University, Milan, Italy

[141] McGill University Health Center, Montreal, QC, Canada;

[142] University of Western Ontario, London, ON, Canada;

[143] Universite Laval, Quebec, QC, Canada;

[144] University of Alberta, Edmonton, AB, Canada;

[145] Dalhousie University, Halifax, NS, Canada;

[146] University of Ottawa, Ottawa, ON, Canada;

[147] University of Montreal, Montreal, QC, Canada;

[148] University of Manitoba, Winnipeg, MB, Canada

[149] University of Montreal Health Center, Montreal, QC, Canada;

[150] Vita-Salute San Raffaele, Milan, Italy;

[151] Martini Clinic Prostate Cancer Center, Hamburg-Eppendorf, Germany

[152] University of Pittsburgh, Pittsburgh, PA, USA

[153] Roswell Park Cancer Institute, Buffalo, NY, USA

[154] University of Montreal Health Center, Montreal, QC, Canada

[155] Martini Clinic Prostate Cancer Center, Hamburg-Eppendorf, Germany

[156] Vita-Salute San Raffaele, Milan, Italy

[157] University of Montreal, Montreal, QC, Canada

[158] Martini Clinic Prostate Cancer Center, Hamburg-Eppendorf, Germany

[159] Vita-Salute San Raffaele, Milan, Italy

[160] Albany Medical Center, Albany, NY, USA

[161] University of Rochester, Rochester, NY, USA

[162] University of Montreal Health Centre, Montreal, QC, Canada

[163] Cancer Prognostics and Health Outcomes Unit, Montreal, QC, Canada

[164] Vita-Salute University San Raffaele, Milan, Italy

[165] University of Montreal Health Centre, Montreal, QC, Canada

[166] Cancer Prognostics and Health Outcomes Unit, Montreal, QC, Canada

[167] Martiniclinic, Prostate Cancer Center University Hospital Hamburg-Eppendorf, Hamburg, Germany

[168] McGill University, Montreal, QC, Canada

[169] University of Montreal Health Center, Dept. of Urology, Montreal, Canada, Montreal, QC, Canada

[170] Department of Urology, Montpellier University Hospital,, Montpellier, France

[171] 4Department of Urology, Prostate Cancer Center Hamburg-Eppendorf, Hamburg, Germany

[172] Cancer Prognosis and Health Outcomes Unit, University of Montreal, Montreal, QC, Canada

[173] Cancer Prognostics and Health Outcomes Unit, University of Montreal Health Centre, Montreal, QC, Canada

[174] Department of Urology, Vita-Salute San Raffaele University,, Milan, Italy

[175] Department of Urology, Prostate Cancer Center Hamburg-Eppendorf,, Hamburg, Germany

[176] University of Rochester, Rochester, NY, USA
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