skip to main content
DOE Patents title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control


Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.

Issue Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
OSTI Identifier:
Patent Number(s):
Application Number:
Los Alamos National Security, LLC
DOE Contract Number:  
Resource Type:
Resource Relation:
Patent File Date: 2010 Nov 05
Country of Publication:
United States

Citation Formats

Cary, Robert E. Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control. United States: N. p., 2015. Web.
Cary, Robert E. Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control. United States.
Cary, Robert E. Tue . "Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control". United States.
title = {Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control},
author = {Cary, Robert E.},
abstractNote = {Highly simplified lateral flow chromatographic nucleic acid sample preparation methods, devices, and integrated systems are provided for the efficient concentration of trace samples and the removal of nucleic acid amplification inhibitors. Methods for capturing and reducing inhibitors of nucleic acid amplification reactions, such as humic acid, using polyvinylpyrrolidone treated elements of the lateral flow device are also provided. Further provided are passive fluid control methods and systems for use in lateral flow assays.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2015},
month = {12}


Save / Share:

Works referenced in this record:

An array of opportunities
journal, April 2002

Identification of the Heme Compound Copurified with Deoxyribonucleic Acid (DNA) from Bloodstains, a Major Inhibitor of Polymerase Chain Reaction (PCR) Amplification
journal, March 1994

  • Akane, Atsushi; Matsubara, Kazuo; Nakamura, Hiroaki
  • Journal of Forensic Sciences, Vol. 39, Issue 2
  • DOI: 10.1520/JFS13607J

Optimal conditions for hybridization with oligonucleotides: A study with myc-oncogene DNA probes
journal, April 1988

  • Albretsen, Catrine; Haukanes, Bjørn-Ivar; Aasland, Rein
  • Analytical Biochemistry, Vol. 170, Issue 1, p. 193-202
  • DOI: 10.1016/0003-2697(88)90108-X

Characterization of a Thermostable UvrD Helicase and Its Participation in Helicase-dependent Amplification
journal, June 2005

  • An, Lixin; Tang, Wen; Ranalli, Tamara A.
  • Journal of Biological Chemistry, Vol. 280, Issue 32, p. 28952-28958
  • DOI: 10.1074/jbc.M503096200

A rapid biosensor for viable B. anthracis spores
journal, August 2004

  • Baeumner, AntjeJ.; Leonard, Barbara; McElwee, John
  • Analytical and Bioanalytical Chemistry, Vol. 380, Issue 1
  • DOI: 10.1007/s00216-004-2726-7

A Universal Nucleic Acid Sequence Biosensor with Nanomolar Detection Limits
journal, February 2004

  • Baeumner, Antje J.; Pretz, Jennifer; Fang, Shirley
  • Analytical Chemistry, Vol. 76, Issue 4
  • DOI: 10.1021/ac034945l

Biosensor for Dengue Virus Detection:  Sensitive, Rapid, and Serotype Specific
journal, March 2002

  • Baeumner, Antje J.; Schlesinger, Nicole A.; Slutzki, Naomi S.
  • Analytical Chemistry, Vol. 74, Issue 6
  • DOI: 10.1021/ac015675e

Biosensors for environmental pollutants and food contaminants
journal, October 2003

The ligase chain reaction in a PCR world.
journal, August 1991

Rapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns
journal, April 1996

Thermodynamic effects of formamide on DNA stability
journal, June 1996

Locked nucleic acid (LNA): fine-tuning the recognition of DNA and RNA
journal, January 2001

Incidence of adamantane resistance among influenza A (H3N2) viruses isolated worldwide from 1994 to 2005: a cause for concern
journal, October 2005

Transmission of the Citrus Variegated Chlorosis Bacterium Xylella fastidiosa with the Sharpshooter Oncometopia nigricans
journal, November 2002

Shape matters
journal, March 2003

  • Buhro, William E.; Colvin, Vicki L.
  • Nature Materials, Vol. 2, Issue 3
  • DOI: 10.1038/nmat844

Signal amplification through nucleotide extension and excision on a dendritic DNA platform
journal, April 2000

Lateral flow microarrays: a novel platform for rapid nucleic acid detection based on miniaturized lateral flow chromatography
journal, April 2007

  • Carter, D. J.; Cary, R. B.
  • Nucleic Acids Research, Vol. 35, Issue 10
  • DOI: 10.1093/nar/gkm269

Helicase structure and mechanism
journal, February 2002

Culture and serological detection of the xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain ofXylella fastidiosa
journal, September 1993

  • Chang, Chung J.; Garnier, Monique; Zreik, Leyla
  • Current Microbiology, Vol. 27, Issue 3
  • DOI: 10.1007/BF01576010

Early diagnosis of bubonic plague using F1 antigen capture ELISA assay and rapid immunogold dipstick
journal, July 2000

  • Chanteau, Suzanne; Rahalison, Lila; Ratsitorahina, Mahery
  • International Journal of Medical Microbiology, Vol. 290, Issue 3, p. 279-283
  • DOI: 10.1016/S1438-4221(00)80126-5

Chemiluminescence Detection for Hybridization Assays on the Flow-Thru Chip, a Three-Dimensional Microchannel Biochip
journal, December 2001

  • Cheek, Brady J.; Steel, Adam B.; Torres, Matthew P.
  • Analytical Chemistry, Vol. 73, Issue 24
  • DOI: 10.1021/ac0108616

Lab-on-a-chip devices for global health: Past studies and future opportunities
journal, January 2007

  • Chin, Curtis D.; Linder, Vincent; Sia, Samuel K.
  • Lab Chip, Vol. 7, Issue 1
  • DOI: 10.1039/B611455E

A nested-PCR assay for detection of Xylella fastidiosa in citrus plants and sharpshooter leafhoppers
journal, March 2004

Multiplex diagnostic platforms for detection of biothreat agents
journal, November 2004

  • Cirino, Nick M.; Musser, Kimberlee A.; Egan, Christina
  • Expert Review of Molecular Diagnostics, Vol. 4, Issue 6
  • DOI: 10.1586/14737159.4.6.841

Purification and characterization of Thermus thermophilus UvrD
journal, February 2003

Nucleic acid sequence-based amplification
journal, March 1991

Synthesis and hybridization of a series of biotinylated oligonucleotides
journal, January 1988

  • Cook, Alan F.; Vuocolo, Edmund; Brakel, Christine L.
  • Nucleic Acids Research, Vol. 16, Issue 9
  • DOI: 10.1093/nar/16.9.4077

Quantitative PCR Method for Diagnosis of Citrus Bacterial Canker
journal, June 2001

Pierce's Disease of Grapevines: Isolation of the Causal Bacterium
journal, January 1978

Identification of A/H5N1 Influenza Viruses Using a Single Gene Diagnostic Microarray
journal, January 2007

  • Dawson, Erica D.; Moore, Chad L.; Dankbar, Daniela M.
  • Analytical Chemistry, Vol. 79, Issue 1
  • DOI: 10.1021/ac061920o

Immobilization of polynucleotides on magnetic particles. Factors influencing hybridization efficiency
journal, September 1991

  • Day, P. J.; Flora, P. S.; Fox, J. E.
  • Biochemical Journal, Vol. 278, Issue 3
  • DOI: 10.1042/bj2780735

Characteristics and Applications of Nucleic Acid Sequence-Based Amplification (NASBA)
journal, January 2002

  • Deiman, Birgit; van Aarle, Pierre; Sillekens, Peter
  • Molecular Biotechnology, Vol. 20, Issue 2
  • DOI: 10.1385/MB:20:2:163

Simultaneous Visual Detection of Multiple Viral Amplicons by Dipstick Assay
journal, August 2005

RNA replication: required intermediates and the dissociation of template, product, and Q.beta. replicase
journal, May 1979

  • Dobkin, Carl; Mills, Donald R.; Kramer, Fred Russell
  • Biochemistry, Vol. 18, Issue 10
  • DOI: 10.1021/bi00577a030

Use of Single Nucleotide Polymorphisms in the plcR Gene for Specific Identification of Bacillus anthracis
journal, April 2005

Optimization of DNA-tagged dye-encapsulating liposomes for lateral-flow assays based on sandwich hybridization
journal, August 2006

  • Edwards, Katie A.; Baeumner, Antje J.
  • Analytical and Bioanalytical Chemistry, Vol. 386, Issue 5
  • DOI: 10.1007/s00216-006-0705-x

Use of laser microdissection greatly improves the recovery of DNA from sperm on microscope slides
journal, October 2003

Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of tomato spotted wilt virus from chrysanthemum
journal, October 2004

An investigation of the rigor of interpretation rules for STRs derived from less than 100 pg of DNA
journal, July 2000

  • Gill, Peter; Whitaker, Jonathan; Flaxman, Christine
  • Forensic Science International, Vol. 112, Issue 1, p. 17-40
  • DOI: 10.1016/S0379-0738(00)00158-4

Biochemical analysis of components of the pre-replication complex of Archaeoglobus fulgidus
journal, August 2003

Detection of foodborne pathogens using DNA probes and a dipstick format
journal, December 1996

Direct fluorescence analysis of genetic polymorphisms by hybridization with oligonucleotide arrays on glass supports
journal, January 1994

  • Guo, Zhen; Guilfoyle, Richard A.; Thiel, Andrew J.
  • Nucleic Acids Research, Vol. 22, Issue 24
  • DOI: 10.1093/nar/22.24.5456

Biochemical Characterization of the DNA Helicase Activity of the Escherichia coli RecQ Helicase
journal, October 2000

  • Harmon, Frank G.; Kowalczykowski, Stephen C.
  • Journal of Biological Chemistry, Vol. 276, Issue 1
  • DOI: 10.1074/jbc.M006555200

Biosensor for the specific detection of a single viable B. anthracis spore
journal, May 2003

  • Hartley, Harriet A.; Baeumner, Antje J.
  • Analytical and Bioanalytical Chemistry, Vol. 376, Issue 3
  • DOI: 10.1007/s00216-003-1939-5

Genetic Diversity of Pierce's Disease Strains and Other Pathotypes of Xylella fastidiosa
journal, February 2001

Populations of Xylella fastidiosa in Plants Required for Transmission by an Efficient Vector
journal, December 1997

Fluorescent Amplified Fragment Length Polymorphism Analysis of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis Isolates
journal, February 2004

Accessing Single Nucleotide Polymorphisms in Genomic DNA by Direct Multiplex Polymerase Chain Reaction Amplification on Oligonucleotide Microarrays
journal, April 2002

  • Huber, Martin; Mündlein, Axel; Dornstauder, Eva
  • Analytical Biochemistry, Vol. 303, Issue 1
  • DOI: 10.1006/abio.2001.5565

Point-of-care diagnostics: will the hurdles be overcome this time?
journal, July 2006

Detection of amantadine-resistant variants among avian influenza viruses isolated in North America and Asia
journal, October 2005

  • Ilyushina, Natalia A.; Govorkova, Elena A.; Webster, Robert G.
  • Virology, Vol. 341, Issue 1, p. 102-106
  • DOI: 10.1016/j.virol.2005.07.003

Development of a multiplex immunocapture RT-PCR assay for detection and differentiation of tomato and tobacco mosaic tobamoviruses
journal, October 1998

Encoded evidence: DNA in forensic analysis
journal, October 2004

  • Jobling, Mark A.; Gill, Peter
  • Nature Reviews Genetics, Vol. 5, Issue 10
  • DOI: 10.1038/nrg1455

Design, biochemical, biophysical and biological properties of cooperative antisense oligonucleotides
journal, January 1995

  • Kandimalla, Ekambar R.; Manning, Adrienne; Lathan, Christopher
  • Nucleic Acids Research, Vol. 23, Issue 17
  • DOI: 10.1093/nar/23.17.3578

DnaB from Thermus aquaticus Unwinds Forked Duplex DNA with an Asymmetric Tail Length Dependence
journal, March 1999

  • Kaplan, Daniel L.; Steitz, Thomas A.
  • Journal of Biological Chemistry, Vol. 274, Issue 11
  • DOI: 10.1074/jbc.274.11.6889

NASBATM isothermal enzymatic in vitro nucleic acid amplification optimized for the diagnosis of HIV-1 infection
journal, December 1991

  • Kievits, Tim; van Gemen, Bob; van Strijp, Dianne
  • Journal of Virological Methods, Vol. 35, Issue 3, p. 273-286
  • DOI: 10.1016/0166-0934(91)90069-C

The total influenza vaccine failure of 1947 revisited: Major intrasubtypic antigenic change can explain failure of vaccine in a post-World War II epidemic
journal, July 2002

  • Kilbourne, E. D.; Smith, C.; Brett, I.
  • Proceedings of the National Academy of Sciences, Vol. 99, Issue 16, p. 10748-10752
  • DOI: 10.1073/pnas.162366899

One-step immobilization of poly(dT)-modified DNA onto non-modified plastic substrates by UV irradiation for microarrays
journal, August 2006

  • Kimura, Naoki
  • Biochemical and Biophysical Research Communications, Vol. 347, Issue 2, p. 477-484
  • DOI: 10.1016/j.bbrc.2006.06.130

Technology platforms for pharmacogenomic diagnostic assays
journal, September 2004

  • Koch, Walter H.
  • Nature Reviews Drug Discovery, Vol. 3, Issue 9
  • DOI: 10.1038/nrd1496

Inclusion of polyvinylpyrrolidone in the polymerase chain reaction reverses the inhibitory effects of polyphenolic contamination of RNA
journal, February 1999

  • Koonjul, P. K.; Brandt, W. F.; Lindsey, G. G.
  • Nucleic Acids Research, Vol. 27, Issue 3
  • DOI: 10.1093/nar/27.3.915

Development of a Novel, Rapid Integrated Cryptosporidium parvum Detection Assay
journal, July 2000

Direct detection of potato leafroll virus in potato tubers by immunocapture and the isothermal nucleic acid amplification method NASBA
journal, June 1997

Current and Developing Technologies for Monitoring Agents of Bioterrorism and Biowarfare
journal, October 2005

Colorimetric detection of immobilised PCR products generated on a solid support
journal, March 1997

Evaluation of NucliSens easyMAG for Automated Nucleic Acid Extraction from Various Clinical Specimens
journal, December 2006

  • Loens, K.; Bergs, K.; Ursi, D.
  • Journal of Clinical Microbiology, Vol. 45, Issue 2
  • DOI: 10.1128/JCM.00894-06

Chromatographic performance of a thin microporous bed of nitrocellulose
journal, November 2001

  • Lönnberg, Maria; Carlsson, Jan
  • Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 763, Issue 1-2, p. 107-120
  • DOI: 10.1016/S0378-4347(01)00376-0

Real-time PCR in the microbiology laboratory
journal, March 2004

Properties of Fluorescent Semiconductor Nanocrystals and their Application to Biological Labeling
journal, December 2001

Molecular Crowding Regulates the Structural Switch of the DNA G-Quadruplex
journal, December 2002

  • Miyoshi, Daisuke; Nakao, Akihiro; Sugimoto, Naoki
  • Biochemistry, Vol. 41, Issue 50
  • DOI: 10.1021/bi020412f

Rapid single-tube immunocapture RT-PCR for the detection of two yam potyviruses
journal, December 1997

journal, November 2003

Capture of Single-Stranded DNA Assisted by Oligonucleotide Modules
journal, January 1998

  • O'Meara, Deirdre; Nilsson, Peter; Nygren, Per-Åke
  • Analytical Biochemistry, Vol. 255, Issue 2
  • DOI: 10.1006/abio.1997.2472

Influenza vaccines: present and future
journal, July 2002

  • Palese, Peter; García-Sastre, Adolfo
  • Journal of Clinical Investigation, Vol. 110, Issue 1
  • DOI: 10.1172/JCI15999

Virulence signatures: microarray-based approaches to discovery and analysis
journal, November 2004

  • Pannucci, James; Cai, Hong; Pardington, Paige E.
  • Biosensors and Bioelectronics, Vol. 20, Issue 4, p. 706-718
  • DOI: 10.1016/j.bios.2004.04.005

DNA analysis with multiplex microarray-enhanced PCR
journal, January 2005

Diagnostic applications of microarrays
journal, August 2006

Microfluidic Device for Rapid (<15 min) Automated Microarray Hybridization
journal, October 2005

DNA Detection Using Recombination Proteins
journal, June 2006

Detection of Xylella fastidiosa in potential insect vectors by immunomagnetic separation and nested polymerase chain reaction
journal, August 1997

Specific PCR detection and identification of Xylella fastidiosa strains causing citrus variegated chlorosis
journal, December 1995

  • Pooler, Margaret R.; Hartung, John S.
  • Current Microbiology, Vol. 31, Issue 6
  • DOI: 10.1007/BF00294703

Microchromatography and microelectrophoresis on nitrocellulose membranes
journal, May 1970

Fate of Pierce's Disease Strains of Xylella fastidiosa in Common Riparian Plants in California
journal, September 1999

Pierce's Disease Bacterium: Mechanism of Transmission by Leafhopper Vectors
journal, November 1979

A novel rapid hybridization technique: paper chromatography hybridization assay (PACHA)
journal, December 1993

Detection and Diversity Assessment of Xylella fastidiosa in Field-Collected Plant and Insect Samples by Using 16S rRNA and gyrB Sequences
journal, July 2003

Amplification and cloning of a long RNA virus genome using immunocapture-long RT-PCR
journal, June 1997

  • Romero, Alicia; Blanco-Urgoiti, Begoña; Ponz, Fernando
  • Journal of Virological Methods, Vol. 66, Issue 1, p. 159-163
  • DOI: 10.1016/S0166-0934(97)00043-8

Advances in Polymerase Chain Reaction on Microfluidic Chips
journal, June 2005

  • Roper, Michael G.; Easley, Christopher J.; Landers, James P.
  • Analytical Chemistry, Vol. 77, Issue 12
  • DOI: 10.1021/ac050756m

Microarray technology – an intellectual property retrospective
journal, September 2003

Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase
journal, January 1988

Dependence of the melting temperature of DNA on salt concentration
journal, April 1965

LNA (locked nucleic acids): synthesis and high-affinity nucleic acid recognition
journal, January 1998

  • Singh, Sanjay K.; Koshkin, Alexei A.; Wengel, Jesper
  • Chemical Communications, Issue 4
  • DOI: 10.1039/A708608C

A Bead-Based Method for Multiplexed Identification and Quantitation of DNA Sequences Using Flow Cytometry
journal, October 2000

Recovery and detection of enterovirus, hepatitis A virus and Norwalk virus in hardshell clams (Mercenaria mercenaria) by RT-PCR methods
journal, February 1999

An Introduction to Monolithic Disks as Stationary Phases for High Performance Biochromatography
journal, January 2000

High-performance membrane chromatography: Highly efficient separation method for proteins in ion-exchange, hydrophobic interaction and reversed-phase modes
journal, September 1993

Membrane chromatography - an integrative concept in the downstream processing of proteins
journal, July 1995

Isothermal reactions for the amplification of oligonucleotides
journal, April 2003

  • Van Ness, J.; Van Ness, L. K.; Galas, D. J.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 8, p. 4504-4509
  • DOI: 10.1073/pnas.0730811100

Helicase-dependent isothermal DNA amplification
journal, August 2004

Potent and nontoxic antisense oligonucleotides containing locked nucleic acids
journal, May 2000

  • Wahlestedt, C.; Salmi, P.; Good, L.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 10
  • DOI: 10.1073/pnas.97.10.5633

Strand displacement amplification—an isothermal, in vitro DNA amplification technique
journal, January 1992

  • Walker, G. Terrance; Fraiser, Melinda S.; Schram, James L.
  • Nucleic Acids Research, Vol. 20, Issue 7, p. 1691-1696
  • DOI: 10.1093/nar/20.7.1691

Using a microfluidic device for 1  l DNA microarray hybridization in 500 s
journal, April 2005

Isolation, Culture, and Pathogenicity of the Bacterium Causing Phony Disease of Peach
journal, January 1983

A highly sensitive immunocapture polymerase chain reaction method for plum pox potyvirus detection
journal, September 1992

PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings
journal, June 2004

Multi-analyte single-membrane biosensor for the serotype-specific detection of Dengue virus
journal, July 2004

  • Zaytseva, NatalyaV.; Montagna, RichardA.; Lee, EunMi
  • Analytical and Bioanalytical Chemistry, Vol. 380, Issue 1
  • DOI: 10.1007/s00216-004-2724-9

Detection of Cell and Tissue Surface Antigens Using Up-Converting Phosphors: A New Reporter Technology
journal, February 1999

  • Zijlmans, H. J. M. A. A.; Bonnet, J.; Burton, J.
  • Analytical Biochemistry, Vol. 267, Issue 1
  • DOI: 10.1006/abio.1998.2965

An amplification-free hybridization-based DNA assay to detect Streptococcus pneumoniae utilizing the up-converting phosphor technology
journal, July 2003

One step visual detection of PCR products with gold nanoparticles and a nucleic acid lateral flow (NALF) device
journal, January 2007

  • Aveyard, Jenny; Mehrabi, Maryam; Cossins, Andrew
  • Chemical Communications, Issue 41
  • DOI: 10.1039/B708859K

Development of a Novel One-Tube Isothermal Reverse Transcription Thermophilic Helicase-Dependent Amplification Platform for Rapid RNA Detection
journal, November 2007

An efficient and economic enhancer mix for PCR
journal, September 2006

  • Ralser, Markus; Querfurth, Robert; Warnatz, Hans-Jörg
  • Biochemical and Biophysical Research Communications, Vol. 347, Issue 3, p. 747-751
  • DOI: 10.1016/j.bbrc.2006.06.151

Developing rapid, point-of-care, multiplex detection for use in lateral flow devices
conference, November 2005

  • Rao, R. S.; Albala, J. S.; Lane, S. L.
  • Optics East 2005, SPIE Proceedings
  • DOI: 10.1117/12.630157

Ligation mediated PCR performed at low denaturation temperatures--PCR melting profiles
journal, September 2003