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標題: 建立磁珠-奈米微脂球檢測技術平臺並應用於快速偵測牛奶中仙人掌桿菌之污染
Development and comparison of three rapid immunomagnetic-bead separation systems for detecting Bacillus cereus in milk, based on different surface-modified immunoliposomal nanovesicles
作者: 朱霈慈
Chu, Pei-Tzu
關鍵字: Bacillus cereus;仙人掌桿菌;immunoliposomal nanovesicles;immunomagnetic beads;免疫奈米微脂體;免疫磁珠
出版社: 食品暨應用生物科技學系所
引用: 1. Hauge S. 1955. Food poisoning caused by aerobic spore-forming bacilli. J Appl Bacterial. 18(3):591-595. 2. Taiwan Department of Health. 2008. Statistic data of food poisoning in Taiwan, Available at Accessed March, 2008. (In Chinese) 3. Schoeni JL, Wong AC. 2005. Bacillus cereus food poisoning and its toxins. J Food Prot. 68(3):636-648. 4. Ehling-Schulz M, Fricker M, Scherer S. 2004. Bacillus cereus, the causative agent of an emetic type of food-borne illness. Mol Nutr Food Res. 48(7):479-487. 5. Agata N, Mori M, Ohta M, Suwan S, Ohtani I, Isobe M. 1994. A novel dodecadepsipeptide, cereulide, isolated from Bacillus cereus causes vacuole formation in HEp-2 cells. FEMS Microbiol Lett. 121:31-34. 6. Granum PE, Lund T. 1997. Bacillus cereus and its food poisoning toxins. FEMS Microbiol Lett. 157:223-228. 7. Granum PE. 1994. Bacillus cereus and its toxins. J Appl Bacteriol Symp Suppl. 23:61S-66S. 8. Beecher DJ, Wong ACL. 1994. Improved purification and characterization of hemolysin BL, a hemolytic dermonecrotic vascular permeability factor from Bacillus cereus. Infect Immun. 62:980-986. 9. Lindback T, Fagerlund A, Rodland MS, Granum PE. 2004. Characterization of the Bacillus cereus Nhe entertoxin. Microbiology. 150:3959-3967. 10. Lund T, De Buyser ML, Granum PE. 2000. A new cytotoxin from Bacillus cereus that may cause necrotic enteritis. Mol Microbiol. 38:254-261. 11. Holmes JR, Plunkett T, Pate P, Roper WL, Alexander WJ. 1981. Emetic food poisoning caused by Bacillus cereus. Arch Intern Med. 141(6):766-767. 12. Suksuwan M. 1983. The incidence of Bacillus cereus in foods in Central Thailand. Southeast Asian J Trop Med Public Health. 14(3):324-329. 13. Morgan SM, Galvin M, Ross RP, Hill C. 2001. Evaluation of a spray-dried lacticin 3147 powder for the control of Listeria monocytogenes and Bacillus cereus in a range of food systems. Lett Appl Microbiol. 33(5):387-391. 14. Nichols GL, Little CL, Mithani V, de Louvois J. 1999. The microbiological quality of cooked rice from restaurants and take-away premises in the United Kingdom. J Food Prot. 62(8):877-882. 15. Kim HJ, Lee DS, Paik HD. 2004. Characterization of Bacillus cereus isolates from raw soybean sprouts. J Food Prot. 67(5):1031-1035. 16. Bartoszewicz M, Hansen BM, Swiecick I. 2008. The members of the Bacillus cereus group are commonly present contaminants of fresh and heat-treated milk. Food Microbiol. 25:588-596. 17. Janntova B, Drabkova M, Vorlova L. 2006. Effect of Bacillus cereus Enzymes on the Milk Quality following Ultra High Temperature Processing. Acta Vet Brno. 75:601-609. 18. Wong HC, Chang MH, Fan JY. 1988. Incidence and characterization of Bacillus cereus isolates contaminating dairy products. Appl Environ Microbiol. 54(3):699-702. 19. Lancette GA, Harmon SM. 1980. Enumeration and confirmation of Bacillus cereus in foods: collaborative study. J Assoc Off Anal Chem. 63(3):581-586. 20. Spira WM, Goepfert JM. 1972. Bacillus cereus-induced fluid accumulation in rabbit ileal loops. Appl Microbiol. 24(3):341-348. 21. Glatz BA, Spira WM, Goepfert JM. 1974. Alteration of vascular permeability in rabbits by culture filtrates of Bacillus cereus and related species. Infect Immun. 10(2):299-303. 22. Melling J, Capel BJ, Turnbull PC, Gilbert RJ. 1976. Identification of a novel enterotoxigenic activity associated with Bacillus cereus. J Clin Pathol. 29(10):938-940. 23. Bergdoll, MS. 1988. Ileal loop fluid accumulation test for diarrheal toxins. Methods Enzymol. 165:306-323. 24. Beecher DJ, Wong AC. 2000. Tripartite haemolysin BL: isolation and characterization of two distinct homologous sets of components from a single Bacillus cereus isolate. Microbiology. 146 (6):1371-1380. 25. Andersson MA, Mikkola R, Helin J, Andersson MC, Salkinoja-Salonen M. 1998. A novel sensitive bioassay for detection of Bacillus cereus emetic toxin and related depsipeptide ionophores. Appl Environ Microbiol. 64(4):1338-1343. 26. Buchanan RL, Schultz FJ. 1994. Comparison of the Tecra VIA kit, Oxoid BCET-RPLA kit and CHO cell culture assay for the detection of Bacillus cereus diarrhoeal enterotoxin. Lett Appl Microbiol. 19(5):353-356. 27. Hsieh YM, Sheu SJ, Chen YL, Tsen HY. 1999. Enterotoxigenic profiles and polymerase chain reaction detection of Bacillus cereus group cells and B. cereus strains from foods and food-borne outbreaks. J Appl Microbiol. 87(4):481-490. 28. Haggblom MM, Apetroaie C, Andersson MA, Salkinoja-Salonen MS. 2002. Quantitative analysis of cereulide, the emetic toxin of Bacillus cereus, produced under various conditions. Appl Environ Microbiol. 68(5):2479-2483. 29. Beecher DJ, Wong AC. 1994. Identification and analysis of the antigens detected by two commercial Bacillus cereus diarrheal enterotoxin immunoassay kits. Appl Environ Microbiol. 60(12):4614-4616. 30. Candish AAG. 1991. Immunological methods in food microbiology. Food Microbiol. 8:1-14. 31. Ramsay G. 1998. DNA chips: State-of-the art. Nature Biotechnol. 16:40-44. 32. Chen CH, Ding HC, Chang TC. 2001. Rapid identification of Bacillus cereus based on the detection of a 28.5-kilodalton cell surface antigen. J Food Prot. 64(3):348-354. 33. Charni N, Perissol C, Le Petit J, Rugani N. 2000. Production and characterization of monoclonal antibodies against vegetative cells of Bacillus cereus. Appl Environ Microbiol. 66(5):2278-2281. 34. Mikami T, Hiraoka K, Murakami T, Boon-Long J, Matsumoto T, Suzuki M. 1990. Detection of common flagella antigen in Bacillus cereus by monoclonal antibody. Microbiol Immunol. 34(8):709-714. 35. Moravek M, Wegscheider M, Schulz A, Dietrich R, Burk C, Martlbauer E. 2004. Colony immunoblot assay for the detection of hemolysin BL enterotoxin producing Bacillus cereus. FEMS Microbiol Lett. 238(1):107-113. 36. Chen CH, Ding HC. 2004. A colony blot immunoassay for the rapid identification of Bacillus cereus. J Food Prot. 67(2):387-390. 37. Malorny B, Tassios PT, Radstrom P, Cook N, Wagner M, Hoorfar J. 2003. Standardization of diagnostic PCR for the detection of foodborne pathogens. Int J Food Microbiol. 83(1):39-48. 38. Nakano S, Maeshima H, Matsumura A, Ohno K, Ueda S, Kuwabara Y, Yamada T. 2004. A PCR assay based on a sequence-characterized amplified region marker for detection of emetic Bacillus cereus. J Food Prot. 67(8):1694-1701. 39. Yang IC, Shih DY, Huang TP, Huang YP, Wang JY, Pan TM. 2005. Establishment of a novel multiplex PCR assay and detection of toxigenic strains of the species in the Bacillus cereus group. J Food Prot. 68(10):2123-2130. 40. Fukushima H, Tsunomori Y, Seki R. 2003. Duplex real-time SYBR green PCR assays for detection of 17 species of food- or waterborne pathogens in stools. J Clin Microbiol. 41(11):5134-5146. 41. Priha O, Hallamaa K, Saarela M, Raaska L. 2004. Detection of Bacillus cereus group bacteria from cardboard and paper with real-time PCR. J Ind Microbiol Biotechnol. 31(4):161-169. 42. Peng H, Ford V, Frampton EW, Restaino L, Shelef LA, Spitz H. 2001. Isolation and enumeration of Bacillus cereus fromfoods on a novel chromogenic plating medium. Food Microbiology. 18(3):231-238. 43. Fermanian C, Wong ACL. 2000. Improved in vitro detection of hemolysin BL from Bacillus cereus. Int J Food Micro. 57(10):1-8. 44. te Giffel MC, Beumer RR, Leijendekkers S, Rombouts FM. 1996. Incidence of Bacillus cereus and Bacillus subtilis in foods in the Netherlands. Food Microbiology. 13(1):53-58. 45. Gray KM, Banada PP, O''Neal E, Bhunia AK. 2005. Rapid Ped-2E9 Cell-Based Cytotoxicity Analysis and Genotyping of Bacillus Species. J Clin Microbiol. 43(12):5865-5872. 46. Day TL, Tatani SR, Notermans S, Bennett RW. 2008. A comparison of ELSA and RPLA for detection of Bacillus cereus diarrhoeal enterotoxin. Journal of Applied Microbiology. 77(1):9-13. 47. Moravek M, Dietrich R, Buerk C, Broussolle V, Guinebretiere MH, Granum PE, Nguyen-The C, Martlbauer E. 2006. Determination of the toxic potential of Bacillus cereus isolates by quantitative enterotoxinanalyses. FEMS Microbiology Letters. 257(2):293-298. 48. Mantynen V, Lindstrom K. 1998. A Rapid PCR-Based DNA Test for Enterotoxic Bacillus cereus. Appl Environ Microbiol. 64(5):1634-1639. 49. Nakano S, Maeshima H, Matsumura A, Ohno K, Ueda S, Kuwabara Y, Yamada T. 2004. A PCR assay based on a sequence-characterized amplified region marker for detection of emetic Bacillus cereus. J Food Prot. 67(8):1694-1701. 50. Bangham AD, Standish MM, Watkins JC. 1965. Diffusion of univalent ions across the lamellae of swollen phospholipids. J Mol Biol. 13(1):238-252. 51. Patel GB, Sprott GD. 1999. Archaeobacterial ether lipid liposomes (archaeosomes) as novel vaccine and drug delivery systems. Crit Rev Biotechnol. 19(4):317-357. 52. Matteucci ML, Thrall DE. 2000. The role of liposomes in drug delivery and diagnostic imaging: a review. Vet Radiol Ultrasound. 41(2):100-107. 53. Banerjee R. 2001. Liposomes: applications in medicine. J Biomater Appl. 16(1):3-21. 54. Bombardelli E. 1991. Phytosome: new cosmetic delivery system. Boll Chim Farm. 130(11):431-438. 55. Weiner N, Lieb L, Niemiec S, Ramachandran C, Hu Z, Egbaria K. 1994. Liposomes: a novel topical delivery system for pharmaceutical and cosmetic applications. J Drug Target. 2(5):405-410. 56. Cevc G. 1996. Transfersomes, liposomes and other lipid suspensions on the skin: permeation enhancement, vesicle penetration, and transdermal drug delivery. Crit Rev Ther Drug Carrier Syst. 13(3-4):257-388. 57. Gibbs BF, Kermasha S, Alli I, Mulligan CN. 1999. Encapsulation in the food industry: a review. Int J Food Sci Nutr. 50(3):213-224. 58. Nelson G. 2002. Application of microencapsulation in textiles. Int J Pharm. 242(1-2):55-62. 59. Vuillemard JC. 1991. Recent advances in the large-scale production of lipid vesicles for use in food products: microfluidization. J Microencapsul. 8(4):547-62. 60. Singh AK, Kilpatrick PK, Carbonell RG. 1996. Application of antibody and fluorophore-derivatized liposomes to heterogeneous immunoassays for d-dimer. Biotechnol Prog. 12(2):272-280. 61. Rongen HA, Bult A, van Bennekom WP. 1997. Liposomes and immunoassays. J Immunol Methods. 204(2):105-133. 62. Lim SJ, Kim CK. 1997. Homogeneous liposome immunoassay for insulin using phospholipase C from Clostridium perfringens. Anal Biochem. 247(1):89-95. 63. Wink T, van Zuilen SJ, Bult A, van Bennekom WP. 1998. Liposome-mediated enhancement of the sensitivity in immunoassays of proteins and peptides in surface plasmon resonance spectrometry. Anal Chem. 70(5):827-832. 64. Meer RR, Park DL. 1995. Immunochemical detection methods for Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes in foods. Rev Environ Contam Toxicol. 142:1-12. 65. Awasthi VD, Garcia D, Klipper R, Goins BA, Phillips WT. 2004. Neutral and Anionic Liposome-Encapsulated Hemoglobin: Effect of Postinserted Poly(ethylene glycol)-distearoylphosphatidylethanolamine on Distribution and Circulation Kinetics. JPET. 309:241-248. 66. Allen TM, Hansen C, Martin F, Redemann C, Yau-Young A. 1991. Liposomes containing synthetic lipid derivatives of poly(ethylene glycol show prolonged circulation half-lives in vivo. Biochim Biophys Acta. 1066(1):29-36. 67. Maruyama K, Takizawa T, Takahashi N, Tagawa T, Nagaike K, Iwatsuru M. 1997. Targeting efficiency of PEG-immunoliposome-conjugated antibodies at PEG terminals. Adv Drug Delivery Rev. 24:235-242. 68. Zrein M, Burckard J, Van Regenmortel MH. 1986. Use of the biotin-avidin system for detecting a broad range of serologically related plant viruses by ELIZA. J Virol Methods. 13(2):121-128. 69. Sakahara H, Saga T. 1999. Avidin-biotin system for delivery of diagnostic agents Adv Drug Deliv Rev. 37(1-3):89-101. 70. Kaasgaard T, Mouritsen OG, Jorgensen K. 2001. Screening effect of PEG on avidin binding to liposome surface receptors. Int J Pharm. 214(1-2):63-65. 71. Hirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP. 2002. Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation. Anal Biochem. 308(2):343-57. 72. Wen HW, Decory TR, Borejsza-Wysocki W, Durst RA. 2006. Investigation of NeutrAvidin-tagged liposomal nanovesicles as universal detection reagents for bioanalytical assays. Talanta. 68(4):1264-1272. 73. Green NM. 1990. Avidin and streptavidin. Methods Enzymol. 184:51-67. 74. Elo HA, Korpela J. 1984. The occurrence and production of avidin: a new conception of the high-affinity biotin-binding protein. Comp Biochem Physiol B. 78(1):15-20. 75. Hendrickson WA, Pahler A, Smith JL, Satow Y, Merritt EA, Phizackerley RP. 1989. Crystal structure of core streptavidin determined from multiwavelength anomalous diffraction of synchrotron radiation. Proc Natl Acad Sci U S A. 86(7):2190-2194. 76. Chaiet L, Wolf FJ. 1964. The properties of streptavidin, a biotin-binding protein produced by streptomycetes. Arch Biochem Biophys. 106:1-5. 77. Green NM. 1975. Avidin. Adv Protein Chem. 29:85-133. 78. Anderson Borge G.I, Skeie M, Sorhaug T, Langsrud T, Granum PE. 2001. Growth and toxin profiles of Bacillus cereus isolated from different food sources. J. Food Microbiology. 69:237-246. 79. Hughes BJ, Kennel S, Lee R, Huang L. 1989. Monoclonal antibody targeting of liposomes to mouse lung in vivo. Cancer Res. 49: 6214-6220. 80. Wang CY, Hiang L. 1987. pH-sensitive immunoliposomes mediate target-cell-specific delivery and controlled expression of a foreign gene in mouse. Proc Natl Acad Sci. U.S.A. 84, pp. 7. 81. Ho RJ, Huang L. 1985. Interactions of antigen-sensitized liposomes with immobilized antibody: a homogeneous solid-phase immunoliposome assay. J. Immunol. 134 :4035- 4040. 82. DeCory TR, Durst RA, Zimmerman SJ, Garringer LA, Paluca G, DeCory HH, Montagna RA. 2005. Development of an immunomagnetic bead-immunoliposome fluorescence assay for rapid detection of Escherichia coli O157:H7 in aqueous samples and comparison of the assay with a standard microbiological method. Appl Environ Microbiol. 71(4):1856-1864. 83. Lasch L, Weissing V, Brandl M. 2003. In: Torchilin VP, Weissig V (eds) Liposomes: a practical approach. Oxford University Press, New York, pp 3-29. 84. Plant AL, Brizgys MV, Locasio-Brown L, Durst RA. 1989. Generic liposome reagent for immunoassays. Anal Biochem. 176(2):420-426. 85. Olsvik O, Popovic T, Skjerve E, Cudjoe KS, Hornes E, Ugelstad J, Uhlen M. 1994. Magnetic separation techniques in diagnostic microbiology. Clin Microbiol Rev. 7(1):43-54. 86. Park S, Oh S, Durst RA. 2004. Immunoliposomes sandwich fluorometric assay (ILSF) for detection of Escherichia coli O157:H7. J Food Sci. 69:M151-M156. 87. Ho JA, Zeng SC, Huang MR, Kuo HY. 2006. Development of Liposomal Immunosensor for the Measurement of Insulin with Femtomole Detection. Anal Chim Acta. 556:127-132. 88. Chen L, Deng L, Liu L, Peng Z. 2007. Immunomagnetic separation and MS/SPR end-detection combined procedure for rapid detection of Staphylococcus aureus and protein A. Biosensors and Bioelectronics. 22: 1487-1492. 89. Murphy M, Carroll A, Walsh C, Whyte P, O''Mahony M, Anderson W, McNamara E, Fanning S. 2007. Development and assessment of a rapid method to detect Escherichia coli O26, O111 and O157 in retail minced beef. Int. J. Hyg. Environ.-Health. 210:155-161. 90. Safarik I, Safarikova M, Forsythe SJ. 1995. The application of magnetic separations in applied microbiology. J Appl Bacteriol. 78:575-585. 91. Liu YJ, Yao DJ, Chang HY, Liu CM, Chen C. 2008. Magnetic bead-based DNA detection with multi-layers quantum dots labeling for rapid detection of Escherichia coli O157:H7. Biosens Bioelectron. 24(4):558-565. 92. Lee HJ, Kim BC, Kim KW, Kim YK, Kim J, Oh MK. 2009. A sensitive method to detect Escherichia coli based on immunomagnetic separation and real-time PCR amplification of aptamers. Biosens Bioelectron. 24(12):3550-3555. 93. de Leon L, Siverio F, Rodriguez A. 2006. Detection of Clavibacter michiganensis subsp. michiganensis in tomato seeds using immunomagnetic separation. J Microbiol Methods. 67(1):141-149. 94. Alefantis T, Grewal P, Ashton J, Khan AS, Valdes JJ, Del Vecchio VG. 2004. A rapid and sensitive magnetic bead-based immunoassay for the detection of staphylococcal enterotoxin B for high-through put screening. Mol Cell Probes. 18(6):379-382. 95. Gessler F, Hampe K, Schmidt M, Bohnel H. 2006. Immunomagnetic beads assay for the detection of botulinum neurotoxin types C and D. Diagn Microbiol Infect Dis. 56(3):225-232. 96. Meyer MH, Stehr M, Bhuju S, Krause HJ, Hartmann M, Miethe P, Singh M, Keusgen M. 2007. Magnetic biosensor for the detection of Yersinia pestis. J Microbiol Methods. 68(2):218-24. 97. Liu GM, Yali H, Li X, Song S. 2006. Applicability of a rapid metod based on immunomagnetic captunt PCR assay for Campylobacter jejuni. Food Control. 17(7):527-532. 98. Yang SY, Lien KY, Huang KJ, Lei HY, Lee GB. 2008. Micro flow cytometry utilizing a magnetic bead-based immunoassay for rapid virus detection. Biosens Bioelectron. 24(4):861-868. 99. Lee WC, Lien KY, Lee GB, Lei HY. 2008. An integrated microfluidic system using magnetic beads for virus detection. Diagn Microbiol Infect Dis. 60(1):51-58. 100. Sakudo A, Baba K, Tsukamoto M, Sugimoto A, Okada T, Kobayashi T, Kawashita N, Takagi T, Ikuta K. 2009. Anionic polymer, poly(methyl vinyl ether-maleic anhydride)-coated beads-based capture of human influenza A and B virus. Bioorg Med Chem. 17(2):752-757. 101. Barletta J, Bartolome A, Constantine NT. 2009. Immunomagnetic quantitative immuno-PCR for detection of less than one HIV-1 virion. J Virol Methods. 157(2):122-132. 102. Schopf E, Fischer NO, Chen Y, Tok JB. 2008. Sensitive and selective viral DNA detection assay via microbead-based rolling circle amplification. Bioorg Med Chem Lett. 18(22):5871-5874. 103. Luke S, Kaul K. 1998. Detection of breast cancer cells in blood using immunomagnetic bead selection and reverse transcription-polymerase chain reaction. Mol Diagn. 3(3):149-155. 104. Chung TH, Chang JY, Lee WC. 2009. Application of magnetic poly(styrene-glycidyl methacrylate) microspheres for immunomagnetic separation of bone marrow cells. J Mag Mag Mater. 321(10):1635-1638. 105. Sun Y, Bi N, Song DQ, Bai Y, Wang LY, Zhang HQ. 2008. Preparation of titania sol-gel matrix for the immunoassay by SPR biosensor with magnetic beads. Sen Actu B. 134(2):566-572. 106. Centi S, Messina G, Tombelli S, Palchetti I, Mascini M. 2008. Different approaches for the detection of thrombin by an electrochemical aptamer-based assay coupled to magnetic beads. Biosens Bioelectron. 23(11):1602-1609. 107. Kim JI, Wang C, Kuizon S, Xu J, Barengolts D, Gray PC, Rubenstein R. 2005. Simple and specific detection of abnormal prion protein by a magnetic bead-based immunoassay coupled with laser-induced fluorescence spectrofluorometry. J Neuroimmunol. 158(1-2):112-119. 108. Nagasaki Y, Kobayashi H, Katsuyama Y, Jomura T, Sakura T. 2007. Enhanced immunoresponse of antibody-mixed-PEG co-immobilized surface construction of high-performance immunomagnetic ELISA system. J Colloid Interface Sci. 309(2):524-530. 109. Centi S, Laschi S, Mascini M. 2007. Improvement of analytical performances of a disposable electrochemical immunosensor by using magnetic beads. Talanta. 73(2):394-399. 110. Lin YY, Liu G, Wai CM, Lin Y. 2007. Magnetic beads-based bioelectrochemical immunoassay of polycyclic aromatic hydrocarbons. Electro Comm. 9(7):1547-1552. 111. S Helali, C Martelet, A Abdelghani, MA Maaref, N Jaffrezic-Renault. 2006. A disposable immunomagnetic electrochemical sensor based on functionalised magnetic beads on gold surface for the detection of atrazine. Anal Chim Acta. 24 (15):5182-5186. 112. Tudorache M, Tencaliec A, Bala C. 2008. Magnetic beads-based immunoassay as a sensitive alternative for atrazine analysis. Talanta. 2(15):839-843. 113. Li H, Li Z, Liu T, Xiao X, Peng Z, Deng L. 2008. A novel technology for biosorption and recovery hexavalent chromium in wastewater by bio-functional magnetic beads. Bioresour Technol. 99(14):6271-6279. 114. Wu PC, Su CH, Cheng FY, Weng JC, Chen JH, Tsai TL, Yeh CS, Su WC, Hwu JR, Tzeng Y, Shieh DB. 2008. Modularly assembled magnetite nanoparticles enhance in vivo targeting for magnetic resonance cancer imaging. Bioconjug Chem. 19(10):197-1979. 114. Bjork I, Petersson BA, Sjoquist J. 1972. Some physiochemical properties of protein A from Staphylococcus aureus. Eur J Biochem. 29(3):579-584. 116. Wen HW, Borejsza-Wysocki W, DeCory TR, Durst RA. 2005. Applications of NeutrAvidin-tagged liposomal nanovesicles in bioassays. Anal Bioanal Chem. 382:1217-1226. 117. Bartlett GR. 1959. Phosphorous assay in column chromatoraphy. J Biol Chem. 234:466-468. 118. Shpigel E, Goldlust A, Eshel A, Ber IK, Efroni G, Singer Y, Levy I, Dekel M, Shoseyov O. 2000. Expression, purification and applications of staphylococcal Protein A fused to cellulose-binding domain. Biotechnol Appl Biochem. 31:197-203. 119. Edwards KA, Baeumner AJ. 2006. Optimization of DNA-tagged liposomes for use in microtiter plate analyses. Anal Bioanal Chem. 386:1613-1623. 120. Du H, Chandaroy P and Hui SW. 1997. Grafted poly-(ethylene glycol) on lipid surfaces inhibits protein adsorption and cell adhesion. Biochim Biophys Acta. 1326: 236-248. 121. Bradley AJ, Devine DV, Ansell SM, Janzen J, Brooks DE. 1998. Inhibition of liposome-induced complement activation by incorporated poly(ethylene glycol)-lipids. Arch Biochem Biophys. 357:185-194. 122. Luppa PB, Sokoll LJ, Chan DW. 2001. Immunosensors-principles and applications to clinical chemistry. Clin Chim Acta. 314(1-2):1-26. 123. Gizeli E, Lowe CR. 1996. Immunosensors. Curr Opin Biotechnol. 7(1):66-71
Bacillus cereus is a major food-born pathogen in Taiwan and its major syndromes include vomiting, fever and diarrhea. To minimize the possibility of exposing consumers to pathogenic B. cereus, this study develops three rapid and sensitive assays that utilize immunomagnetic beads (IMBs) and different immunoliposomal nanovesicles (IMLNs) systems. In these assays, anti-B. cereus antibody-conjugated IMBs were applied to capture B. cereus in samples; fluorescent dyes-loaded IMLNs were employed to increase the detection signal. Hence, a sandwich complex was formed as “IMBs-B. cereus-IMLNs”. In the first system, antibodies were directly conjugated to the liposomal surface. The optimal IMLNs had a diameter of 300 nm conjugated 0.25 mol% antibodies. The limit of detection (LOD) of this first system was 5 CFU/mL of B. cereus in milk samples. For increasing the distance between antibodies and liposomal nanovesicles, the second system antibody was tagged on the far end of polyethylene glycol (PEG) chains on the liposomal surface. The optimal PEG-IMLN had a diameter of 200 nm conjugated 0.5 mol% antibodies. The LOD of this second assay was closed to the LOD of the first assay as 5 CFU/mL of B. cereus in milk. In order to decrease the LOD in milk, we further designed the third system, a new type of PEG-IMLN, which surface was modified by 1 mol% PEG chains and 0.5 mol% biotins. Successfully, the Biotin-PEG-IMLN detection system could reach to the 2 CFU/mL of B. cereus about 6 hours in milk. To evaluate the specificity of these assays, nine Gram positive and negative bacteria were tested and no significant interference was found. Conclusively, this study elucidates the feasibility of using a novel IMB/IMLN assay to detect less than 5 CFU/mL of B. cereus within one working day.

仙人掌桿菌是台灣地區引起食物中毒的主要病原菌之一,感染時造成的中毒症狀包括嘔吐,發燒和腹瀉。為了保障消費者食的安全,本研究利用不同表面修飾的免疫奈米微脂體(IMLNs),建立了三種針對仙人掌桿菌快速且靈敏的免疫磁珠(IMBs)偵測平台。分析樣品時,先藉由在磁珠表面修飾上仙人掌桿菌抗體的免疫磁珠從樣品中分離出仙人掌桿菌,接著加入包埋螢光染劑的免疫奈米微脂體增加偵測訊號。在第一個檢測系統中,抗體直接地修飾在奈米微脂體表面;最適化結果為粒徑 300 nm 免疫微脂體上修飾 0.25 mol%的抗體。在牛奶樣品中,最低偵測極限(LOD)為 5 CFU/mL 仙人掌桿菌。而在第二個檢測系統中,將抗體標定在微脂體表面PEG的尾端上,以增加抗體和奈米微脂體的間距並增強其擺動性;而系統的最適化條件為 200 nm 的 PEG-IMLN上修飾0.5 mol% 抗體。在牛奶樣品中,第二個檢測平臺的LOD為 5 CFU/mL 的仙人掌桿菌。而為了降低檢測平臺在牛奶的LOD,我們進一步的設計了第三個檢測系統,以一種新型態的 PEG-IMLN,其表面修飾了1 mol% 的PEG和0.5 mol% 生物素。我們利用生物素標定的免疫微脂體檢測系統可成功的在 6小時內檢測出牛奶中的仙人掌桿菌,其LOD達到 2 CFU/mL。 為評估此三種檢測平臺的專一性,藉由測試九株革蘭氏陽性和陰性菌,結果顯示本實驗所建立的檢測平臺具有良好的專一性。本研究藉由不同表面修飾的免疫奈米微脂球發展出一種快速檢測牛奶當中仙人掌桿菌之方法。
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