DOCSLIB.ORG

Duke University Dissertation Template

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Duke University Dissertation Template On-chip Labeling via Surface Initiated Enzymatic Polymerization (SIEP) for Nucleic Acids Hybridization Detection by Vinalia Tjong Department of Biomedical Engineering Duke University Date:_______________________ Approved: ___________________________ Ashutosh Chilkoti, Supervisor ___________________________ Stefan Zauscher ___________________________ William Reichert ___________________________ Gabriel Lopez ___________________________ Aimee Zaas Dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Biomedical Engineering in the Graduate School of Duke University 2013 i v ABSTRACT On-chip Labeling via Surface Initiated Enzymatic Polymerization (SIEP) for Nucleic Acids Hybridization Detection by Vinalia Tjong Department of Biomedical Engineering Duke University Date:_______________________ Approved: ___________________________ Ashutosh Chilkoti, Supervisor ___________________________ Stefan Zauscher ___________________________ William Reichert ___________________________ Gabriel Lopez ___________________________ Aimee Zaas An abstract of a dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Biomedical Engineering in the Graduate School of Duke University 2013 i v Copyright by Vinalia Tjong 2013 Abstract Current techniques for nucleic acid analysis often involve extensive sample preparation that requires skilled personnel and multiple purification steps. In this dissertation, we introduce an on-chip, isothermal, post-hybridization labeling and signal amplification technique that can directly interrogate unmodified DNA and RNA samples on a microarray format, eliminating the need for microarray sample pre- processing. We name this technique Surface Initiated Enzymatic Polymerization (SIEP), where we exploit the ability of a template independent DNA polymerase called Terminal Deoxynucleotidyl Transferase (TdT) to catalyze the formation of long single- stranded DNA (ssDNA) chain from the 3’-end of a short DNA primer, which is tethered on the surface, and TdT’s ability to incorporate unnatural reporter nucleotides, such as fluorescent nucleotides. We hypothesize that polymerization of a long ssDNA chain while incorporating multiple fluorescent nucleotides on target DNA or RNA hybridized to probe printed on a surface will provide a simple and powerful, isothermal method for on-chip labeling and signal amplification. We developed the SIEP methodology by first characterizing TdT biochemical reaction to polymerize long homopolymer ssDNA (> 1000 bases) starting from the 3’-OH of ten bases oligonucleotides. We found that the preferred monomers (deoxynucleotide, dNTP) are dATP and dTTP, and that the length of the ssDNA extension is determined iv by the ratio of input monomer (dNTP) to initiator (short oligonucleotides). We also investigated TdT’s ability to incorporate fluorescent dNTPs into a ssDNA chain by examining the effect of the molar ratios of fluorescent dNTP to natural dNTP on the initiation efficiency, the degree of fluorophore incorporation, the length and the polydispersity of the polymerized DNA strand. These experiments allowed us to incorporate up to ~50 fluorescent Cy3-labeled dNTPs per kilobase into a ssDNA chain. With the goal of using SIEP as an on-chip labeling method, we also quantified TdT mediated signal amplification on the surface by immobilizing ssDNA oligonucleotide initiators on a glass surface followed by SIEP of DNA. The incorporation of multiple fluorophores into the extended DNA chain by SIEP translated to a up to ~45 fold increase in signal amplification compared to the incorporation of a single fluorophore. SIEP was then employed to detect hybridization of DNA (25 bases), short miRNA (21 bases) and long mRNA (1400 bases) by the post-hybridization, on-chip polymerization of fluorescently labeled ssDNA that was grown from the 3’-OH of hybridized target strands. A dose-response curve for detection of DNA hybridization by SIEP was generated, with a ~1 pM limit of detection (LOD) and a 2-log linear dynamic range while the detection of short miRNA and fragmented mRNA targets resulted in ~2 pM and ~10 pM LOD, respectively with a 3-log linear dynamic range. We further developed SIEP for colorimetric detection by exploiting the presence of negatively charged phosphate backbone on the surface as target DNA or RNA v hybridizes on the immobilized probe. The net negative charge on the surface is further increased by TdT catalyzed polymerization of long ssDNA. We then used positively charged gold nanoparticles as reporters, which can be further amplified through electroless metallization, creating DNA spots that are visible by eye. We observed an increase of 100 fold in LOD due to SIEP amplification. Overall, we demonstrated the use of SIEP methodology to label unmodified target DNA and RNA on chip, which can be detected through fluorescence signal or colorimetric signal of metallized DNA spots. This methodology is straightforward and versatile, is compatible with current microarray technology, and can be implemented using commercially available reagents. vi To Mama, Kasey, and Aina vii Contents Abstract ......................................................................................................................................... iv List of Tables .............................................................................................................................. xiii List of Figures .............................................................................................................................xiv Acknowledgements ................................................................................................................. xvii 1. Motivation and Background ............................................................................................... 1 1.1 Nucleic acid analysis: molecular diagnostic promises and challenges ..................... 1 1.2 Nucleic acid target amplification ................................................................................... 3 1.2.1 Polymerase chain reaction (PCR) .............................................................................. 3 1.2.2 Isothermal amplification techniques ........................................................................ 5 1.3 Signal amplification: nucleic acid labeling and probes ............................................... 7 1.3.1 Fluorescence-based probes and signal amplification ............................................. 8 1.3.2 Affinity-based signal amplification .......................................................................... 8 1.3.3 Highly branched DNA signal amplification ........................................................... 9 1.3.4 Nanoparticle-based probes and signal amplification........................................... 11 1.4 Microarray chip technology .......................................................................................... 12 1.4.1 Nucleic acid target amplification and labeling in microarray ............................ 13 1.4.2 On-chip signal amplification strategies .................................................................. 15 2. Surface Initiated Enzymatic Polymerization (SIEP) ...................................................... 18 2.1 DNA as a polymeric material ....................................................................................... 18 2.2 Polymerization: chemical synthesis vs DNA polymerization ................................. 22 2.3 Surface initiated polymerization (SIP)......................................................................... 25 viii 2.4 SIEP for nucleic acid analysis ....................................................................................... 27 3. Characterization of Terminal Deoxynucleotidyl Transferase (TdT) ........................... 31 3.1 Biochemical properties of TdT ...................................................................................... 31 3.1.1 Reaction stoichiometry of DNA polymerization catalyzed by TdT ................... 32 3.1.2 Initiator: primer requirement ................................................................................... 33 3.1.3 Monomer: natural and unnatural dNTP ................................................................ 33 3.1.4 Metal ion as cofactor ................................................................................................. 34 3.2 TdT applications in molecular biology ........................................................................ 34 3.3 Characterization of TdT reaction for SIEP technology: formation of long homopolymer (≥1Kb) catalyzed by TdT ........................................................................... 36 3.3.1 Effects of initiator-monomer composition ............................................................. 37 3.3.2 Effects of monomer to initiator ratio ....................................................................... 41 3.3.3 Non-processivity of TdT: implication of stepwise polymerization of homopolymer ssDNA chain ............................................................................................. 43 3.4 Incorporation of unnatural nucleotides ...................................................................... 45 3.4.1 Incorporation of fluorescent dNTPs ....................................................................... 46 3.4.2 Incorporation of amino allyl-dUTP .......................................................................
Load more

Recommended publications
  • Open Research Online Oro.Open.Ac.Uk
    Open Research Online The Open University’s repository of research publications and other research outputs The Role of Viral Load in the Pathogenesis of HIV-2 Infection in West Africa Thesis How to cite: Ariyoshi, Koya (1998). The Role of Viral Load in the Pathogenesis of HIV-2 Infection in West Africa. PhD thesis The Open University. For guidance on citations see FAQs. c 1998 Koya Ariyoshi https://creativecommons.org/licenses/by-nc-nd/4.0/ Version: Version of Record Link(s) to article on publisher’s website: http://dx.doi.org/doi:10.21954/ou.ro.000101f4 Copyright and Moral Rights for the articles on this site are retained by the individual authors and/or other copyright owners. For more information on Open Research Online’s data policy on reuse of materials please consult the policies page. oro.open.ac.uk THE ROLE OF VIRAL LOAD D< THE PATHOGENESIS OF HTV-2 INFECTION IN WEST AFRICA BY KOYAARIYOSHI MRC Laboratories, Fajara, The Gambia, West Africa ^ ew • I A thesis submitted to the Open University in fulfilment for the degree of Doctor of Philosophy 1998 Collaborating Establishments: University College Medical School (London) Statens Serum Institute (Copenhagen) Institute of Molecular Medicine (Oxford) Institute of Cancer Research (London) _ ProQuest Number:C706741 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion.
    [Show full text]
  • New and Emerging Yeast Pathogens KEVIN C
    CLINICAL MICROBIOLOGY REVIEWS, Oct. 1995, p. 462–478 Vol. 8, No. 4 0893-8512/95/$04.0010 Copyright q 1995, American Society for Microbiology New and Emerging Yeast Pathogens KEVIN C. HAZEN* Division of Clinical Microbiology, Department of Pathology, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908 INTRODUCTION .......................................................................................................................................................462 DEFINITION OF NEW OR EMERGING YEAST PATHOGENS ......................................................................462 WHICH YEASTS ARE NEW OR EMERGING PATHOGENS? .........................................................................463 ANATOMIC SITES ATTACKED BY YEASTS.......................................................................................................464 HISTOPATHOLOGY .................................................................................................................................................466 TREATMENT OF INFECTIONS DUE TO UNUSUAL YEASTS .......................................................................466 Catheter Removal ...................................................................................................................................................466 Antifungal Therapy.................................................................................................................................................469 MICROBIOLOGICAL IDENTIFICATION ............................................................................................................469
    [Show full text]
  • Helicobacter Pylori
    Advanced Techniques in Diagnostic Microbiology Yi-Wei Tang Charles W. Stratton Advanced Techniques in Diagnostic Microbiology Yi-Wei Tang Charles W. Stratton Molecular Infectious Disease Laboratory Clinical Microbiology Laboratory Vanderbilt University Medical Center Vanderbilt University Medical Center Nashville, TN 37232-5310 Nashville, TN 37232-5310 USA USA [email protected] [email protected] Library of Congress Control Number: 2005935335 ISBN-10: 0-387-29741-3 e-ISBN 0-387-32892-0 ISBN-13: 978-0387-29741-5 Printed on acid-free paper. C 2006 Springer Science+Business Media, LLC. All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. Printed in the United States of America. (TB/EB) 987654321 springer.com Contributors Jaber Aslanzadeh Ali Danesh Division of Clinical Department of Experimental Microbiology Therapeutics Department of Pathology University Health Network Hartford Hospital and Clinical 200 Elizabeth Street Laboratory Partners Toronto, Ontario, Canada M5G 2C4 85 Seymour Street Hartford, CT 06102, USA Diane Dare Research Development Unit George Bolton Manchester Metropolitan BD Biosciences University 10975 Torreyana Road St.
    [Show full text]
  • Pharmacoeconomics
    PharmacoEconomics The costs of diagnosing and treating sexually transmitted infections in low- and middle- income countries from 2006 to 2014: An updated systematic review --Manuscript Draft-- Manuscript Number: Full Title: The costs of diagnosing and treating sexually transmitted infections in low- and middle- income countries from 2006 to 2014: An updated systematic review Article Type: Systematic Review Funding Information: United States Agency for International Not applicable Development (AID-674-A-12-00029) Abstract: Background: Sexually transmitted infections (STIs) are co-factors for HIV infection and can cause significant morbidity. Expanding on a prior systematic review, we aimed to summarize recent literature on the costs of diagnosing and treating curable STIs in low- and middle-income countries (LMICs). Methods: We conducted a systematic review using pre-established search strategies. Citations were eligible if published between 1 January 2006 and 31 December 2014 and if they contained provider-perspective cost information reflective of STI-related service provision in LMICs. We extracted all cost values and used regression analysis to explore determinants. Cost drivers were analyzed thematically. Results: We identified 44 articles for inclusion; 24 (54.6%) represented Sub-Saharan Africa. We extracted 202 cost values; 72 (35.6%) characterized syndromic management approaches, 57 (28.2%) mobile outreach services. Syphilis was a common focus (70 (34.7%)). Sixty-five (32.2%) cost values represented cost- effectiveness measures as compared to simple unit costs. The median for all cost values was (USD 2015) $10.90 (cost-effectiveness measures $115.88; unit costs $4.15). Regression analysis indicated that cost effective measures were lower in Africa than other continents.
    [Show full text]
  • Quantitation of Cytomegalovirus: Methodologic Aspects and Clinical Applications
    CLINICAL MICROBIOLOGY REVIEWS, July 1998, p. 533–554 Vol. 11, No. 3 0893-8512/98/$04.0010 Copyright © 1998, American Society for Microbiology. All Rights Reserved. Quantitation of Cytomegalovirus: Methodologic Aspects and Clinical Applications 1 2 MICHAEL BOECKH * AND GUY BOIVIN Fred Hutchinson Cancer Research Center, Seattle, Washington,1 and Centre Hospitalier de l’Universite´ Laval, Sainte-Foy, Quebec, Canada2 INTRODUCTION .......................................................................................................................................................534 PATHOPHYSIOLOGY OF CMV DISSEMINATION ...........................................................................................534 METHODS FOR CMV QUANTIFICATION..........................................................................................................534 Quantitative Viral Cultures...................................................................................................................................535 Principle and assay characteristics..................................................................................................................535 Assay performance..............................................................................................................................................535 Reproducibility and test accuracy ....................................................................................................................535 Summary ..............................................................................................................................................................535
    [Show full text]
  • Point-Of-Care Diagnostics for Lung Transplantation
    Point-of-Care Diagnostics for Lung Transplantation by Andrew Thomas Sage A thesis submitted in conformity with the requirements for the degree of Doctor of Philosophy Graduate Department of Pharmaceutical Sciences University of Toronto © Copyright by Andrew Thomas Sage 2015 Point-of-Care Diagnostics for Lung Transplantation Andrew Thomas Sage Doctor of Philosophy Graduate Department of Pharmaceutical Sciences University of Toronto 2015 Abstract Over the last few decades, lung transplantation (LTx) has become a well-established therapy for patients suffering from end-stage lung disease. Despite the many innovations that have occurred in recent years, the outcomes following LTx still lag behind those of other organ transplant procedures due to a lack of technologies that could monitor predictive biomarkers. Recently, several genomic and proteomic analytes have been identified that are prognostic of LTx outcome; however, traditional analytical methods are impractical for use in the transplant setting. Rapid analysis of molecular biomarkers reporting on the initial and long-term status of a donated organ would improve utilization rates and transplant outcomes. The work herein focuses on developing a novel class of chip-based sensors that would enable direct analysis of mRNAs and proteins that are correlated with the suitability of a lung for transplant. The biosensing approach utilized in these studies was performed using a simple and elegant electrochemical reporter system with metal electrodes of varying morphologies and sizes. ii By fabricating fractal circuit sensors (FraCS), we demonstrated the technological and biological validation required for a point-of-care (POC) diagnostic device to be used for gene quantitation in the transplant setting.
    [Show full text]
  • Attachment 1 Statewide Standard List of Abbreviations
    Statewide Standard List of Abbreviations Revised June 28, 2016 Attachment 1 Statewide Standard List of Abbreviations Table of Contents Table 1: Abbreviations, Acronyms and Symbols ............................................................. 2 Table 2: Do NOT Use Abbreviations ............................................................................. 25 1 Statewide Standard List of Abbreviations Revised June 28, 2016 Table 1: Abbreviations, Acronyms and Symbols Abbreviation or Acronym Definition (L) left (R) right /hpf per high power field ā before A and O alert and oriented A and P auscultation and percussion aa of each AACRN Advanced AIDS Certified Registered Nurse AAHIVM American Academy of HIV medicine AB abortion abd abdomen ac before meals ACHES Abdominal pain, chest pain, headache, eye problems, and severe leg pains (early danger signs of oral contraceptive adverse effects) ACRN AIDS Certified Registered Nurse ACTG AIDS Clinical Trial Group ADA American Dietetic Association ADAP AIDS Drug Assistance Program ADC AIDS Dementia Complex ADLs Activities of daily living AED automated external defibrillator AETC AIDS Education and Training Centers AF anteflexed AFB acid-fast bacilli AGCUS atypical glandular cells of undetermined significance AGN Acute glomerulonephritis AHYD Adolescent Health and Youth Development AIDS acquired immunodeficiency syndrome AIN anal intraepithelial neoplasia AlkPhos alkaline phosphatase ALT alanine aminotransferase 2 Statewide Standard List of Abbreviations Revised June 28, 2016 amb ambulatory AMBU air-shields
    [Show full text]
  • Branched Dna Nanostructures for Molecular Diagnostics
    BRANCHED DNA NANOSTRUCTURES FOR MOLECULAR DIAGNOSTICS A Dissertation Presented to the Faculty of the Graduate School of Cornell University in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy by Mark Richard Hartman August 2013 © 2013 Mark Richard Hartman ALL RIGHTS RESERVED ii BRANCHED DNA NANOSTRUCTURES FOR MOLECULAR DIAGNOSTICS Mark Richard Hartman, Ph.D. Cornell University 2013 DNA nanotechnology has been enormously successful in using DNA as a nanoscale construction material, and consistent progress in using increasingly complex DNA- mediated assemblies and designs has been reported in the literature. However, the field of DNA nanostructures has sometimes been lacking for concrete applications. Meanwhile, DNA has also been widely used as probes and primers for nucleic acid diagnostics applications, and this provides an extremely critical application with potential for great medical benefits, and these DNA probes and primers are readily interfaced with DNA nanotechnology. Thus, in this Dissertation, I discuss the combination of (1) DNA in its role as a recognition element for nucleic acid detection, and (2) DNA as a structural material for nanoscale self-assembly. This combination represents a fusion of “structure” and “function”, and will ultimately lead to great advancements in the field of nucleic acid diagnostics. iii BIOGRAPHICAL SKETCH I received my B.S. degree in Chemical and Biomolecular Engineering from Cornell University in 2007. As an undergraduate with a minor in Biological Engineering, I worked with Prof. Dan Luo in the Biological and Environmental Engineering on the design and synthesis of DNA nanostructures and portable detection using DNA nanobarcodes. Upon graduation, I remained at Cornell and entered the Masters of Engineering program (2007 – 2008), where I worked on DNA-mediated self-assembly of gold nanoparticles.
    [Show full text]
  • Lennette's Laboratory Diagnosis of Viral
    Lennette’s Laboratory Diagnosis of Viral Infections NFECT I OUS I SEASE Fourth Edition Fourth I D Edition AN D THERAPY SER I ES About the book Volume 50 • Written from the perspective of the diagnostician, this bestselling book is the definitive text on the laboratory diagnosis of human viral diseases. • Contains a wealth of illustrations, tables, and algorithms to enhance your understanding of Viral Infections Viral of Lennette’s Lennette’s of this ever-evolving field. • A ready reference source for virologists, microbiologists, epidemiologists, laboratorians, infectious disease specialists and students. Laboratory Unique features • Has a new syndromic approach and discusses the differential diagnosis of potential causative agents along with suggestions for the appropriate diagnostic response. Diagnosis of • Examines the field’s rapid changes in technology, the continuing emergence of new viruses, and the newly described viral etiologies for clinical syndromes. Laboratory Diagnosis • Explores the increasingly important subject of molecular techniques in detail, covering the design of molecular tests, the importance of genotyping and viral Viral Infections sequence analysis, and the use of microarrays in diagnostic virology. Reviews of previous editions Fourth Edition a worthy addition to the bookshelves of the specialised virus laboratory as it contains a wealth of information not readily available elsewhere. The Journal of Clinical Pathology a comprehensive and valuable addition to any medical library. It will appeal to all students of virology, medical microbiology and infectious diseases. The information is set out in a user-friendly fashion and allows the reader to obtain pertinent information without being deluged with too many data [...] recommend[ed] highly.
    [Show full text]
  • Etude De Determinants De La Transmission Du Vih De La Mere a L’Enfant Au Burkina Faso
    UNIVERSITE LIBRE DE BRUXELLES ANNEE 2003-2004 THESE Pour obtenir le grade académique de DOCTEUR EN SCIENCES AGRONOMIQUES ET INGENIERIE BIOLOGIQUE FACULTE DES SCIENCES Par Mr Olivier MANIGART ETUDE DE DETERMINANTS DE LA TRANSMISSION DU VIH DE LA MERE A L’ENFANT AU BURKINA FASO Jury : Directeur de thèse : Mr le Professeur Philippe VAN DE PERRE, co- Directrice de thèse : Mme le Professeur Carine VAN LINT, Président : Mr le Professeur Georges HUEZ, Mme le Professeur Véronique HALLOIN, Mr le Professeur Nicolas MEDA, Mr le Professeur John WERENNE, Mr le Professeur Arsène BURNY, Mr le Professeur Philippe LEPAGE. 1 Couverture : route Ouagadougou – Bobo-Dioulasso, juillet 2003. 2 A ma petite fille adorée, Isia, 3 4 Je me suis réveillé, tiré de la lourde torpeur d’une sieste de saison sèche par des cris stridents et des acclamations saccadées qui provenaient du devant de ma cour. Je me suis levé lourdement et j’ai écarté le pagne qui protège de la lumière et de la chaleur tellement vives en cette période, pour découvrir que les « Masques » étaient descendus dans la rue. Au contraire, la lumière était diffuse et sinistre et la lourdeur de l’air était telle qu’une angoisse profonde m’étreignit instantanément. En général, les « Masques » sortent lors de périodes rituelles – ce qui n’était pas le cas – ou, au cours de phénomènes étranges. Ce sont les initiés qui portent le masque et qui fustigent à tout-vas : c’est souvent l’occasion de corrections exemplaires pour ceux qui se sont mal comportés… ou de vengeances personnelles. J’ai levé les yeux pour observer le ciel : il était obscurci d’un halo de nuages circulaire d’un diamètre de ce qui semblait être plusieurs kilomètres.
    [Show full text]
  • Mycology Proficiency Testing Program
    Mycology Proficiency Testing Program Test Event Critique May 2014 Table of Contents Mycology Laboratory 2 Mycology Proficiency Testing Program 3 Test Specimens & Grading Policy 5 Test Analyte Master Lists 7 Performance Summary 9 Commercial Device Usage Statistics 11 Yeast Descriptions 12 Y-1 Candida parapsilosis 12 Y-2 Candida glabrata 15 Y-3 Rhodotorula minuta 18 Y-4 Trichosporon asahii 21 Y-5 Prototheca wickerhamii 24 Antifungal Susceptibility Testing - Yeast 27 Antifungal Susceptibility Testing - Mold (Educational) 29 1 Mycology Laboratory Mycology Laboratory at the Wadsworth Center, New York State Department of Health (NYSDOH) is a reference diagnostic laboratory for the fungal diseases. The laboratory services include testing for the dimorphic pathogenic fungi, unusual molds and yeasts pathogens, antifungal susceptibility testing including tests with research protocols, molecular tests including rapid identification and strain typing, outbreak and pseudo-outbreak investigations, laboratory contamination and accident investigations and related environmental surveys. The Fungal Culture Collection of the Mycology Laboratory is an important resource for high quality cultures used in the proficiency-testing program and for the in-house development and standardization of new diagnostic tests. Mycology Proficiency Testing Program provides technical expertise to NYSDOH Clinical Laboratory Evaluation Program (CLEP). The program is responsible for conducting the Clinical Laboratory Improvement Amendments (CLIA)-compliant Proficiency Testing (Mycology) for clinical laboratories in New York State. All analytes for these test events are prepared and standardized internally. The program also provides continuing educational activities in the form of detailed critiques of test events, workshops and occasional one-on-one training of laboratory professionals. Mycology Laboratory Staff and Contact Details Name Responsibility Phone Email Director (on leave of Dr.
    [Show full text]
  • | Hao Hata Kama Maria Del Mat Man at Ka Na Malo Matter
    |HAO HATA KAMA MARIAUS009772330B2 DEL MAT MAN AT KA NA MALO MATTER (12 ) United States Patent (10 ) Patent No. : US 9 ,772 ,330 B2 Nussbaum ( 45 ) Date of Patent : Sep . 26 , 2017 (54 ) NUCLEIC ACID APTAMER -BASED WO 2007024676 A2 3 / 2007 DIAGNOSTIC METHODS WITH NOVEL Wo 2007117444 A2 10 / 2007 TECHNIQUES FOR SIGNAL WO 2009070640 A2 6 / 2009 ENHANCEMENT OTHER PUBLICATIONS (71 ) Applicant : APTATECK BIO LTD . , Ness Ziona ( IL ) Zeeshan M , et al .“ Comparison of different phenotypic methods of detection of methicillin resistance in Staphylococcus aureus with (72 ) Inventor: Ofer Nussbaum , Rehovot ( IL ) the molecular detection of mec - a gene ” J . Coll . Physicians. Surg . Pak . 17 :666 -70 (2007 ) . ( 73 ) Assignee : APTATECK BIO LTD ., Rehovot ( IL ) Edmonds, M . “ Branched RNA ” BioEssays . 6 :212 - 216 ( 1987 ). Lin ( I ) ( Clinical Chemistry 2009 vol . 55 , p . 1686 - 1693 ) . ( * ) Notice : Subject to any disclaimer , the term of this Goddard et al. ( Prog Polym Sci 2007 vol. 32 , p . 698 -725 ) . patent is extended or adjusted under 35 Lisa R . Paborsky et al . “ The Single - stranded DNA Aptamer -binding U . S . C . 154 (b ) by 0 days . Site of Human Thrombin ” The Journal of Biological Chemistry . 268 :20606 - 20811 ( 1993 ) . ( 21) Appl. No. : 14 / 565 ,759 Mandona Sassanfar, Jack W . Szostak . “ An RNA motif that binds ATP ” Letters to Nature . 364 : 550 - 553 ( 1993 ) . ( 22 ) Filed : Dec. 10 , 2014 Monica Mir, Ioanis Katakis . “ Aptamers as elements of bioelectronic devices " The Royal Society of Chemistry . 3 :620 -622 (2007 ) . (65 ) Prior Publication Data Matthew Levy et al . “ Quantum - Dot Aptamer Beacons for the Detection of Proteins ” ChemBioChem .
    [Show full text]