Summary
Clinical and environmental analyses frequently necessitate rapid, simple, and inexpensive point-of-care or field tests. These semiquantitative tests may be later followed up by confirmatory laboratory-based assays, but can provide an initial scenario assessment important for resource mobilization and threat confinement. Lateral-flow assays (LFAs) and dip-stick assays, which are typically antibody-based and yield a visually detectable signal, provide an assay format suiting these applications extremely well. Signal generation is commonly obtained through the use of colloidal gold or latex beads, which yield a colored band either directly proportional or inversely proportional to the concentration of the analyte of interest. Here, dye-encapsulating liposomes as an alternative are discussed. The LFA biosensors described in this chapter rely on the sandwich-hybridization of a nucleic acid sequence-based amplified (NASBA) mRNA target between a membrane immobilized capture probe and a visible dye (sulforhodamine B)-encapsulating liposome conjugated reporter probe. Although the methodology of this chapter is focused on LFAs for the detection of RNA through sandwich hybridization, the information within can be readily adapted for sandwich and competitive immunoassays. Included are an introduction and application notes toward this end. These include notes ranging from the detection of nonamplified RNA and single-stranded DNA, conjugation protocols for antibodies and other proteins to liposomes, and universal assay formats.
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References
Helbig, J. H., Luck, P. C., Kunz, B., and Bubert, A. (2006) Evaluation of the Duopath Legionella lateral flow assay for identification of Legionella pneumophila and Legionella species culture isolates. Appl Environ Micro-biol. 72, 4489–4491
Koide, M., Haranaga, S., Higa, F., Tateyama, M., Yamane, N., and Fujita, J. (2007) Comparative evaluation of Duopath Legionella lateral flow assay against the conventional culture method using Legionella pneumophila and Legionella anisa strains. Jpn J Infect Dis. 60, 214–216
http://www.kit.nl/biomedical_research/assets/images/Lepto_lateral_flow_protocol.doc
Johnston, S. P., Ballard, M. M., Beach, M. J., Causer, L., and Wilkins, P. P. (2003) Evaluation of three commercial assays for detection of Giardia and Cryptosporidium organisms in fecal specimens. J Clin Microbiol. 41, 623–626
Ketema, F., Zeh, C., Edelman, D. C., Saville, R., and Constantine, N. T. (2001) Assessment of the performance of a rapid, lateral flow assay for the detection of antibodies to HIV. J Acquir Immune Defic Syndr. 27, 63–70
Cazacu, A. C., Demmler, G. J., Neuman, M. A., Forbes, B. A., Chung, S. Y., Greer, J., Alvarez, A. E., Williams, R., and Bartholoma, N. Y. (2004) Comparison of a new lateral-flow chromatographic membrane immunoassay to viral culture for rapid detection and differentiation of influenza A and B viruses in respiratory specimens. J Clin Microbiol. 42, 3661–3664
Mokkapati, V. K., Sam Niedbala, R., Kardos, K., Perez, R. J., Guo, M., Tanke, H. J., and Corstjens, P. L. (2007) Evaluation of UPlink-RSV: prototype rapid antigen test for detection of respiratory syncytial virus infection. Ann N Y Acad Sci. 1098, 476–485
Slinger, R., Milk, R., Gaboury, I., and Diaz-Mitoma, F. (2004) Evaluation of the QuickLab RSV test, a new rapid lateral-flow immunoassay for detection of respiratory syncytial virus antigen. J Clin Microbiol. 42, 3731–3733
van Hengel, A. J., Capelletti, C., Brohee, M., and Anklam, E. (2006) Validation of two commercial lateral flow devices for the detection of peanut proteins in cookies: interlaboratory study. J AOAC Int. 89, 462–468
http://www.brittneylimited.com/PDF%20Files/FDA_Files/Pregnancy-strip.pdf
Edwards, K. A., Curtis, K. L., Sailor, J., Bae-umner, A. (2008) Universal liposomes: Preparation and usage for the detection of mRNA. Anal Bioanal Chem.391, 1689–1702, Epub 2008 March 8
Jung, K., Zachow, J., Lein, M., Brux, B., Sinha, P., Lenk, S., Schnorr, D., and Loen-ing, S. A. (1999) Rapid detection of elevated prostate-specific antigen levels in blood: performance of various membrane strip tests compared. Urology. 53, 155–160
http://www.brittneylimited.com/PDF%20Files/PSA-Prostrate-strip.pdf
Sharma, S. K., Eblen, B. S., Bull, R. L., Burr, D. H., and Whiting, R. C. (2005) Evaluation of lateral-flow clostridium botulinum neuro-toxin detection kits for food analysis. Appl Environ Microbiol. 71, 3935–3941
Park, C. H., Kim, H. J., Hixon, D. L., and Bubert, A. (2003) Evaluation of the duopath verotoxin test for detection of shiga toxins in cultures of human stools. J Clin Microbiol. 41, 2650–2653
Capps, K. L., McLaughlin, E. M., Murray, A. W. A., Aldus, C. F., Wyatt, G. M., Peck, M. W., Amerongen, A., Ariens, R. M. C., Wich-ers, J. H., Baylis, C. L., Wareing, D. R. A., and Bolton, F. J. (2004) Validation of three rapid screening methods for detection of verotoxin-producing Escherichia coli in foods: Interlabo-ratory study. J Aoac Int. 87, 68–77
Smits, H. L., Eapen, C. K., Sugathan, S., Kuria-kose, M., Gasem, M. H., Yersin, C., Sasaki, D., Pujianto, B., Vestering, M., Abdoel, T. H., and Gussenhoven, G. C. (2001) Lateral-flow assay for rapid serodiagnosis of human leptospirosis. Clin Diagn Lab Immunol. 8, 166–169
Kassler, W. J., Dillon, B. A., Haley, C., Jones, W. K., and Goldman, A. (1997) On-site, rapid HIV testing with same-day results and counseling. Aids. 11, 1045–1051
Prevention, U. S. C. f. D. C. a. (1996) in “MMWR Morb Mortal Wkly Rep”, Vol. 45, pp. 468–80
Baeumner, A. J., Jones, C., Wong, C. Y., and Price, A. (2004) A generic sandwich-type biosensor with nanomolar detection limits. Anal Bioanal Chem. 378, 1587–1593
Baeumner, A. J., Pretz, J., and Fang, S. (2004) A universal nucleic acid sequence biosensor with nanomolar detection limits. Anal Chem. 76, 888–894
Seal, J., Braven, H., Wallace, P. (2006) Point-of-care nucleic acid lateral-flow tests. IVD Technol.
Rule, G. S., Montagna, R. A., and Durst, R. A. (1996) Rapid method for visual identification of specific DNA sequences based on DNA-tagged liposomes. Clin Chem. 42, 1206–1209
Corstjens, P. , Zuiderwijk, M., Brink, A., Li, S., Feindt, H., Niedbala, R. S., and Tanke, H. (2001) Use of up-converting phosphor reporters in lateral-flow assays to detect specific nucleic acid sequences: a rapid, sensitive DNA test to identify human papillomavirus type 16 infection. Clin Chem. 47, 1885–1893
Corstjens, P. L., Zuiderwijk, M., Nilsson, M., Feindt, H., Sam Niedbala, R., and Tanke, H. J. (2003) Lateral-flow and up-converting phosphor reporters to detect single-stranded nucleic acids in a sandwich-hybridization assay. Anal Biochem. 312, 191–200
Carney, J., Braven, H., Seal, J., Whitworth, E. (2006) Present and future applications of gold in rapid assays. IVD Technol. 12, 41–49
Oku, Y., Kamiya, K., Kamiya, H., Shibahara, Y., Ii, T., and Uesaka, Y. (2001) Development of oligonucleotide lateral-flow immunoassay for multi-parameter detection. J Immunol Methods. 258, 73–84
Zaytseva, N. V., Montagna, R. A., Lee, E. M., and Baeumner, A. J. (2004) Multi-analyte single-membrane biosensor for the serotype-specific detection of Dengue virus. Anal Bio-anal Chem. 380, 46–53
Corstjens, P. L., Chen, Z., Zuiderwijk, M., Bau, H. H., Abrams, W. R., Malamud, D., Sam Niedbala, R., and Tanke, H. J. (2007) Rapid assay format for multiplex detection of humoral immune responses to infectious disease pathogens (HIV, HCV, and TB). Ann N Y Acad Sci. 1098, 437–445
Zhang, C., Zhang, Y., and Wang, S. (2006) Development of multianalyte flow-through and lateral-flow assays using gold particles and horseradish peroxidase as tracers for the rapid determination of carbaryl and endosulfan in agricultural products. J Agric Food Chem. 54, 2502–2507
Aldus, C. F., Van Amerongen, A., Ariens, R. M., Peck, M. W., Wichers, J. H., and Wyatt, G. M. (2003) Principles of some novel rapid dipstick methods for detection and characterization of verotoxigenic Escherichia coli. J Appl Microbiol. 95, 380–389
O'Farrell, B., Bauer, J. (2006) Developing highly sensitive, more-reproducible lateral-flow assays Part 1: New approaches to old problems. IVD Technol.
Hans, H. B., Karl Pflanz, E. J., and Klewitz, T. M. (2002) Qualification of cellulose nitrate membranes for lateral-flow assays. IVD Technol.
Harlow, E., and Lane, D. (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratories, Cold Spring Harbor
Jones, K. (2001) Membrane immobilization of nucleic acids, Part 2: Probe Attachment Techniques. IVD Technol. September
Sischo, W. M., Atwill, E. R., Lanyon, L. E., and George, J. (2000) Cryptosporidia on dairy farms and the role these farms may have in contaminating surface water supplies in the northeastern United States. Prev Vet Med. 43, 253–267
Bangs Laboratories, I. (1999) Lateral Flow Tests. http://www.bangslabs.com/technotes/303.pdfAccessed
Edwards, K. A., and Baeumner, A. J. (2006) Optimization of DNA-tagged dye-encapsulating liposomes for lateral-flow assays based on sandwich hybridization. Anal Bioanal Chem. 386, 1335–1343
Batteiger, B., Newhall, V. W. J., and Jones, R. B. (1982) The use of Tween 20 as a blocking agent in the immunological detection of proteins transferred to nitrocellulose membranes. J Immunol Methods. 55, 297–307
Jones, K. D. (1999) Troubleshooting protein binding in nitrocellulose membranes Part I: Principles. IVD Technol. 32
Van Dam, A. P., Van den Brink, H. G., and Smeenk, R. J. (1990) Technical problems concerning the use of immunoblots for the detection of antinuclear antibodies. J Immunol Methods. 129, 63–70.
Zampieri, S., Ghirardello, A., Doria, A., Tonello, M., Bendo, R., Rossini, K., and Gambari, P. F. (2000) The use of Tween 20 in immunoblotting assays for the detection of autoantibodies in connective tissue diseases. J Immunol Methods. 239, 1–11
Smits, H. L., Chee, H. D., Eapen, C. K., Kuria-kose, M., Sugathan, S., Gasem, M. H., Yersin, C., Sakasi, D., Lai, A. F. R. F., Hartskeerl, R. A., Liesdek, B., Abdoel, T. H., Goris, M. G., and Gussenhoven, G. C. (2001) Latex based, rapid and easy assay for human leptospirosis in a single test format. Trop Med Int Health. 6, 114–118
Chan, C. P., Sum, K. W., Cheung, K. Y., Glatz, J. F., Sanderson, J. E., Hempel, A., Lehmann, M., Renneberg, I., and Ren-neberg, R. (2003) Development of a quantitative lateral-flow assay for rapid detection of fatty acid-binding protein. J Immunol Methods. 279, 91–100
Wang, S., Zhang, C., Wang, J., Zhang, Y. (2005) Development of colloidal gold-based flow-through and lateral-flow immunoassays for the rapid detection of the insecticide car-baryl. Anal Chim Acta. 546, 161–66
Klewitz, T., Gessler, F., Beer, H., Pflanz, K., and Scheper, T. (2006) Immunochromato-graphic assay for determination of botulinum neurotoxin type D. Sensors and Actuators B: Chemical. 113, 582–589
Shyu, R. H., Shyu, H. F., Liu, H. W., and Tang, S. S. (2002) Colloidal gold-based immunochromatographic assay for detection of ricin. Toxicon. 40, 255–258
Greenwald, R., Esfandiari, J., Lesellier, S., Houghton, R., Pollock, J., Aagaard, C., Andersen, P., Hewinson, R. G., Chambers, M., and Lyashchenko, K. (2003) Improved serodetection of Mycobacterium bovis infection in badgers (Meles meles) using multian-tigen test formats. Diagn Microbiol Infect Dis. 46, 197–203
Birnbaum, S., Uden, C., Magnusson, C. G., and Nilsson, S. (1992) Latex-based thin-layer immunoaffinity chromatography for quantita-tion of protein analytes. Anal Biochem. 206, 168–171
LaBorde, R. and O'Farrell, B. (2002) Paramagnetic-particle detection in lateral-flow assays. IVD Technol. 8, 36–41
Szoka, F., Olson, F., Heath, T., Vail, W., Mayhew, E., and Papahadjopoulos, D. (1980) Preparation of unilamellar lipo-somes of intermediate size (0.1–0.2 μmol) by a combination of reverse phase evaporation and extrusion through polycarbonate membranes. Biochim Biophys Acta. 601, 559–571
Szoka, F., Jr., and Papahadjopoulos, D. (1978) Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation. Proc Natl Acad Sci U S A. 75, 4194–4198
Edwards, K. A. and Baeumner, A. J. (2006) Sequential injection analysis system for the sandwich hybridization-based detection of nucleic acids. Anal Chem. 78, 1958–1966
Rule, G. S., Montagna, R. A., and Durst, R. A. (1997) Characteristics of DNA-tagged liposomes allowing their use in capillary-migration, sandwich-hybridization assays. Anal Biochem. 244, 260–269
Rongen, H. A. H., Bult, A., and vanBen-nekom, W. P. (1997) Liposomes and immu-noassays. J Imm Meth. 204, 105–133
Esch, M. B., Baeumner, A. J., and Durst, R. A. (2001) Detection of Cryptosporidium par-vum using oligonucleotide-tagged liposomes in a competitive assay format. Anal Chem. 73, 3162–3167
Locascio-Brown, L., Plant, A. L., Chesler, R., Kroll, M., Ruddel, M., and Durst, R. A. (1993) Liposome-based flow-injection immunoassay for determining theophylline in serum. Clin Chem. 39, 386–391
Edwards, K. A. and Baeumner, A. J. (2007) Synthesis of a liposome incorporated 1-car-boxyalkylxanthine–phospholipid conjugate and its recognition by an RNA aptamer. Talanta. 71, 365–372
Edwards, K. A., and Baeumner, A. J. (2006) Optimization of DNA-tagged liposomes for use in microtiter plate analyses. Anal Bioanal Chem. 386, 1613–1623
Goral, V. N., Zaytseva, N. V., and Baeumner, A. J. (2006) Electrochemical microfluidic biosensor for the detection of nucleic acid sequences. Lab Chip. 6, 414–421
Deiman, B., van Aarle, P., and Sillekens, P. (2002) Characteristics and applications of nucleic acid sequence-based amplification (NASBA). Mol Biotechnol. 20, 163–179
Clemons, W. M., Jr., May, J. L., Wimberly, B. T., McCutcheon, J. P., Capel, M. S., and Ramakrishnan, V. (1999) Structure of a bacte rial 30S ribosomal subunit at 5.5 A resolution. Nature. 400, 833–840
Cook, N. (2003) The use of NASBA for the detection of microbial pathogens in food and environmental samples. J Microbiol Methods. 53, 165–174
Birch, L., Dawson, C. E., Cornett, J. H., and Keer, J. T. (2001) A comparison of nucleic acid amplification techniques for the assessment of bacterial viability. Lett Appl Microbiol. 33, 296–301
Keer, J. T., and Birch, L. (2003) Molecular methods for the assessment of bacterial viability. J Microbiol Methods. 53, 175–183
Vandamme, A. M., Van Dooren, S., Kok, W., Goubau, P., Fransen, K., Kievits, T., Schmit, J. C., De Clercq, E., and Desmyter, J. (1995) Detection of HIV-1 RNA in plasma and serum samples using the NASBA amplification system compared to RNA-PCR. J Virol Methods. 52, 121–32
Simpkins, S. A., Chan, A. B., Hays, J., Popping, B., and Cook, N. (2000) An RNA transcription-based amplification technique (NASBA) for the detection of viable Salmonella enterica. Lett Appl Microbiol. 30, 75–79
Sakallah, S. A. (2000) Molecular diagnostics of infectious diseases: state of the technology. Biotechnol Annu Rev. 6, 141–61
Iqbal, S. S., Mayo, M. W., Bruno, J. G., Bronk, B. V., Batt, C. A., and Chambers, J. P. (2000) A review of molecular recognition technologies for detection of biological threat agents. Biosens Bioelectron. 15, 549–578
Wolfe, H. J. (1988) DNA probes in diagnostic pathology. Am J Clin Pathol. 90, 340–344
Mac Kenzie, W. R., Hoxie, N. J., Proctor, M. E., Gradus, M. S., Blair, K. A., Peterson, D. E., Kazmierczak, J. J., Addiss, D. G., Fox, K. R., Rose, J. B., and et al. (1994) A massive outbreak in Milwaukee of crypt-osporidium infection transmitted through the public water supply. N Engl J Med. 331, 161–167
Fayer, R., Trout, J. M., Lewis, E. J., Santin, M., Zhou, L., Lal, A. A., and Xiao, L. (2003) Contamination of Atlantic coast commercial shellfish with Cryptosporidium. Parasitol Res. 89, 141–145
Nime, F. A., Burek, J. D., Page, D. L., Hols-cher, M. A., and Yardley, J. H. (1976) Acute enterocolitis in a human being infected with the protozoan Cryptosporidium. Gastroenter-ology. 70, 592–598
Brown, T. A. (2002) Genomes, Wiley, New York
Somer, L., and Kashi, Y. (2003) A PCR method based on 16S rRNA sequence for simultaneous detection of the genus Listeria and the species Listeria monocytogenes in food products. J Food Prot. 66, 1658–1665
Fuchs, B. M., Glockner, F. O., Wulf, J., and Amann, R. (2000) Unlabeled helper oligo-nucleotides increase the in situ accessibility to 16S rRNA of fluorescently labeled oligonu-cleotide probes. Appl Environ Microbiol. 66, 3603–3607
Ban, N., Nissen, P., Hansen, J., Capel, M., Moore, P. B., and Steitz, T. A. (1999) Placement of protein and RNA structures into a 5 A-resolution map of the 50S ribosomal subu-nit. Nature. 400, 841–847
Nugen, S. R., Leonard, B., and Baeumner, A. J. (2006) Application of a unique server-based oligonucleotide probe selection tool toward a novel biosensor for the detection of Streptococcus pyogenes. Biosens Bioelectron. 22, 2442–2448
Small, J., Call, D. R., Brockman, F. J., Straub, T. M., and Chandler, D. P. (2001) Direct detection of 16S rRNA in soil extracts by using oligonucleotide microarrays. Appl Environ Microbiol. 67, 4708–4716
Chaney, R., Rider, J., and Pamphilon, D. (1999) Direct detection of bacteria in cellular blood products using bacterial ribosomal RNA-directed probes coupled to electrochem-iluminescence. Transfus Med. 9, 177–188
Chen, C. S., Baeumner, A. J., and Durst, R. A. (2005) Protein G-liposomal nanovesicles as universal reagents for immunoassays. Talanta. 67, 205–211
Plant, A. L., Brizgys, M. V., Locasio-Brown, L., and Durst, R. A. (1989) Generic liposome reagent for immunoassays. Anal Biochem. 176, 420–426
Edwards, K. A., and Baeumner, A. J. (2007) DNA-oligonucleotide encapsulating lipo-somes as a secondary signal amplification means. Anal Chem. 79, 1806–1815
Zuker, M. (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31, 3406–3415
Altschul, S. F., Gish, W., Miller, W., Myers, E. W., and Lipman, D. J. (1990) Basic local alignment search tool. J Mol Biol. 215, 403–410
Benson, D. A., Karsch-Mizrachi, I., Lipman, D. J., Ostell, J., and Wheeler, D. L. (2004) Gen-Bank: update. Nucleic Acids Res. 32, D23–D26
Koppel, D. E. (1972) Analysis of macromo-lecular polydispersity in intensity correlation spectroscopy – method of cumulants. J Chem Phys. 57, 4814–4820
Frisken, B. J. (2001) Revisiting the method of cumulants for the analysis of dynamic light-scattering data. Appl Opt. 40, 4087–4091
Fiske, C. H., and Subbarow, Y. (1925) The colorimetric determination of phosphorus. J Biol Chem. 66, 375–400
(2001) RNeasy Mini Handbook, Qiagen
Boom, R., Sol, C. J., Salimans, M. M., Jansen, C. L., Wertheim-van Dillen, P. M., and van der Noordaa, J. (1990) Rapid and simple method for purification of nucleic acids. J Clin Micro-biol. 28, 495–503
Bartlett, G. R. (1959) Phosphorus assay in column chromatography. J Biol Chem. 234, 466–468
Coyne, V., James, D., Reid, S., and Rybicki, E. (1998) Detection of nucleic acids by hybridization. http://www.mcb.uct.ac.za/hybridn.htm Accessed March 24 2007
Boutet, V., Delaunay, V., De Oliveira, M. C., Boquet, D., Grognet, J. M., Grassi, J., and Deverre, J. R. (2000) Real-time monitoring of the hybridization reaction: application to the quantification of oligonucleotides in biological samples. Biochem Biophys Res Commun. 268, 92–98
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The authors gratefully acknowledge the review of this chapter by Kit Meyers and Barbara Leonard.
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Edwards, K.A., Baeumner, A.J. (2009). Liposome-Enhanced Lateral-Flow Assays for the Sandwich-Hybridization Detection of RNA. In: Rasooly, A., Herold, K.E. (eds) Biosensors and Biodetection. Methods in Molecular Biology™, vol 504. Humana Press. https://doi.org/10.1007/978-1-60327-569-9_13
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