Patent SG-11201809927X-A (SG)
Title: Improvements in or relating to nucleic acid amplification processes
Inventor: PROVINS JARROD
IPC: C12Q1/68
Filed: 2017-06-30
Abstract: INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property :::` , MD 1101111 0 DOI HIM 011101 0 101111111111111111111111111111111111111 0111111 Organization International Bureau (10) International Publication Number 03 (43) International Publication Date .....•\"\" WO 2018/002649 Al 04 January 2018 (04.01.2018) W I PO I PCT (51) International Patent Classification: (GB). SHEN, Daiwei; c/o LumiraDx UK Ltd, 3 More Lon- C12Q 1/68 (2006.01) BOIL 7/00 (2006.01) don Riverside, London Greater London SE1 2AQ (GB). KRAYNACK, Bryan; c/o LumiraDx UK Ltd, 3 More Lon- (21) International Application Number: PCT/GB2017/051927 don Riverside, London Greater London SE1 2AQ (GB). (22) International Filing Date: (74) Agent: LIPSCOMBE, Martin John et al.; Nash Matthews 30 June 2017 (30.06.2017) LLP, 24 Hills Road, Cambridge Cambridgeshire CB2 1JP (GB). (25) Filing Language: English (81) Designated States (unless otherwise indicated, for every (26) Publication Language: English kind of national protection available): AE, AG, AL, AM, (30) Priority Data: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, 1611469.6 30 June 2016 (30.06.2016) GB CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, (71) Applicant: LUMIRADX UK LTD [GB/GB]; 3 More Lon- HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, don Riverside, London Greater London SE1 2AQ (GB). 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, (72) Inventors: PROVINS, Jarrod; c/o LumiraDx UK Ltd, 3 OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, More London Riverside, London Greater London SE1 2AQ (54) : steps of: a duplex structure by; (b) causing a nick at each of said nicking sites in the strands of the duplex; (c) using a polymerase to extend so as and (c) as desired so as to cause the production of multiple copies of the newly synthesised nucleic acid; characterised (57) 71' hybridisation C temperature Title: IMPROVEMENTS IN OR RELATING TO NUCLEIC ACID AMPLIFICATION PROCESSES comprising permit a the formation amplification the nicked steps in that the amplification = = Average STAR -0.5C Rate, Chlamydia trachomatis gDNA 55000 — = 45000 —:: 8 . 2.3 35000 —:: 13 .. 8 . 5.-z 25000 —:: x, - 60 ______ ------- ----------------- 55 — = = 50 = • ••••• • lt Temperature 'Z.) • • .\" il. , :. .— 40 . . : - E : 35 ' _ • : 30 • — 25 = Target 100 copies M=18) Target 10 copies (n=17) —No Target Control (n=12) = _ = F, \" s • • 2 15000 3 —,:•:•:•:. . D7 5000 —i: ... . . .. _ = _ = = — -5000 —** 0 ----- • - ••••••••.•-• -• . . .....,.... ... — ..............„.........................„................••.................... 4 5 6 7 8 9 10 20 Time (minutes) _ = at FIG. 8 Disclosed is a method of performing a non-isothermal nucleic acid amplification reaction, the method (a) mixing a target sequence with one or more complementary single stranded primers in conditions which event in which the primers hybridise to the target, which hybridisation event, directly or indirectly, leads to comprising two nicking sites disposed at or near opposite ends of the duplex; and performing an to form newly synthesised nucleic acid, which extension with the polymerase recreates nicking sites; (d) repeating which the method is performed is non-isothermal, and subject to a reduction of at least 2° C during the ,-, 01 the ei of © process --..... strands * 1-1 1:) (b) ei C [Continued on next page] WO 2018/002649 Al 0 111 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). Published: — with international search report (Art. 21(3)) — with amended claims (Art. 19(1)) process of steps (b)-(d).