Patent SG-11201908594V-A (SG)
Title: Nanomaterial-based bacterial sensors
Inventor: SUTARLIE LAURA
IPC: C12Q1/10
Filed: 2018-03-27
Abstract: Negativelycharged risDNA-AuNPs (assay component ) Positively .charged luminescent mates - Eels (essay comps ::ent 2) Without bacteria Legend: (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (43) International Publication Date 04 October 2018 (04.10.2018) WIPO I PCT onion °nolo OH 1101 H 010 11011111° oimIE (10) International Publication Number WO 2018/182510 Al (51) International Patent Classification: C12Q 1/06 (2006.01) GO1N 21/64 (2006.01) C12Q 1/10 (2006.01) G01N 33/569 (2006.01) (21) International Application Number: PCT/SG2018/050137 (22) International Filing Date: 27 March 2018 (27.03.2018) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 10201702528R 28 March 2017 (28.03.2017) SG (71) Applicant: AGENCY FOR SCIENCE, TECHNOLO- GY AND RESEARCH [SG/SG]; 1 Fusionopolis Way, #20-10, Connexis North Tower, Singapore 138632 (SG). (72) Inventors: SUTARLIE, Laura; c/o Institute of Materials Research and Engineering, 2 Fusionopolis Way, #08-03 In- novis, Singapore 138634 (SG). OW, Sian Yang; c/o Insti- tute of Materials Research and Engineering, 2 Fusionopo- lis Way, #08-03 Innovis, Singapore 138634 (SG). CHEN, Shilin; c/o Institute of Materials Research and Engineering, 2 Fusionopolis Way, #08-03 Innovis, Singapore 138634 (SG). SU, Xiaodi; c/o Institute of Materials Research and Engineering, 2 Fusionopolis Way, #08-03 Innovis, Singa- pore 138634 (SG). (74) Agent: SPRUSON & FERGUSON (ASIA) PTE LTD; P.O. Box 1531, Robinson Road Post Office, Singapore 903031 (SG). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, (54) Title: NANOMATERIAL-BASED BACTERIAL SENSORS dsDNA-AuNPs Fluorescence materials Quenched f u ore sce ce materials Bacteria cell (57) : Nanomaterial-based Bacterial Sensors This invention relates to a sensor that detects bacteria cells comprising (a) a primary negatively charged, nanoparticulate sensing material; (b) a secondary positively charged, fluorescent sensing material; (c) a housing; and (d) at least one illuminator; wherein said housing contains said primary negatively charged, nanoparticulate sensing ma- terial, said secondary positively charged fluorescent sensing material and a sample potentially comprising bacteria in aqueous medium, wherein said illuminator provides light of at least one pre-specified wavelength to excite at least said secondary positively charged, ?d fluorescent material, wherein said secondary positively charged, fluorescent material electrostatically attached to bacteria cells provides at least one fluorescent response at a second different wavelength kn wherein both i and n are integers, wherein said negatively charged, nanoparticulate sensing material electrostatically attached to said fluorescent material suppresses fluorescing of said fluorescent mate- [Continued on next page] WO 2018/182510 Al MIDEDIM011010EIREEM0MOHE11111111111011111111111111111 DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: — of inventorship (Rule 4.17 (iv)) Published: with international search report (Art. 21(3)) in black and white; the international application as filed contained color or greyscale and is available for download from PATENTSCOPE rial at said second wavelength kn; and wherein said housing permits illumination of the contents of said housing by said illuminator and wherein said housing further permits the detection of a fluorescent response at a second wavelength kn. The negatively charged material includes (dsDNA coated) spherical AuNPs and graphene oxide (GO). The positively charged fluorescent material includes water soluble cationic conjugated polyelectrolytes (CCPE) or positively charged peptide/polymer labeled with fluorescence dye. The sensor makes use of the FRET phenomenon between the primary and secondary sensing materials. The sensor allows making a dis- tinction between living and dead bacteria and can measure the total bacteria count. A method for detecting bacteria utilizing the sensor is another part of the invention.