Nghiên cứu tổng hợp vật liệu ceo2 có cấu trúc nano nhằm ứng dụng trong cảm biến ADN

BỘ GIÁO DỤC VÀ ĐÀO TẠO TRƯỜNG ĐẠI HỌC BÁCH KHOA HÀ NỘI NGUYỄN THỊ NGUYỆT NGHIÊN CỨU TỔNG HỢP VẬT LIỆU CeO2 CÓ CẤU TRÚC NANO NHẰM ỨNG DỤNG TRONG CẢM BIẾN ADN LUẬN ÁN TIẾN SĨ KHOA HỌC VẬT LIỆU Hà Nội - 2020 BỘ GIÁO DỤC VÀ ĐÀO TẠO TRƯỜNG ĐẠI HỌC BÁCH KHOA HÀ NỘI NGUYỄN THỊ NGUYỆT NGHIÊN CỨU TỔNG HỢP VẬT LIỆU CeO2 CÓ CẤU TRÚC NANO NHẰM ỨNG DỤNG TRONG CẢM BIẾN ADN Ngành: Khoa học vật liệu Mã số: 9440122 LUẬN ÁN TIẾN SĨ KHOA HỌC VẬT LIỆU NGƯỜI HƯỚNG DẪN KHOA HỌC: PGS.TS PHƯƠNG ĐÌNH TÂM GS.TS TRẦN TRUNG Hà Nội - 2020 LỜI CẢM ƠN Trải qua năm học tập nghiên cứu hết mình, tác giả hoàn thành luận án Tiến sĩ Khoa học Vật liệu Viện AIST, trường Đại học Bách Khoa Hà Nội Để hồn thành luận án này, tác giả nhận hướng dẫn tận tâm, động viên bảo hết lòng hai thầy hướng dẫn PGS TS Phương Đình Tâm GS TS Trần Trung Qua đây, tác giả xin bày tỏ lòng biết ơn sâu sắc tới hai thầy đặc biệt PGS TS Phương Đình Tâm – người ln bên cạnh, sát sao, bảo, góp ý bước thực luận án tác giả Bên cạnh đó, tác giả nhận tư vấn, giúp đỡ, động viên vô to lớn thầy PGS TS Phạm Hùng Vượng, TS Nguyễn Đức Dũng PGS TS Đặng Thị Thanh Lê tồn thể thầy cô, bạn nghiên cứu sinh học viên cao học theo học Viện AIST anh chị em, bạn bè đồng nghiệp Tác giả xin gửi lời cảm ơn sâu sắc tới tồn thể thầy anh chị em bạn! Trong thời gian theo học Viện AIST, trường Đại học Bách Khoa Hà Nội, tác giả nhận tận tình giúp đỡ tập thể lãnh đạo Viện AIST, phòng ban chức trường Đại học Bách Khoa Hà Nội Tác giả xin trân trọng cảm ơn tất giúp đỡ này! Ngoài ra, tác giả xin tỏ lòng biết ơn sâu sắc tới đại gia đình nội ngoại hai bên, cảm ơn chồng tác giả đồng hành, tiếp sức cho tác giả suốt thời gian thực luận án! Tác giả Nguyễn Thị Nguyệt i LỜI CAM ĐOAN Tơi xin cam đoan cơng trình nghiên cứu riêng Các kết nêu luận án trung thực chưa tác giả khác công bố Hà Nội, ngày TM Tập thể hướng dẫn tháng năm 2020 Tác giả PGS.TS Phương Đình Tâm Nguyễn Thị Nguyệt ii MỤC LỤC LỜI CẢM ƠN i LỜI CAM ĐOAN ii MỤC LỤC iii DANH MỤC CÁC KÝ HIỆU VÀ CHỮ VIẾT TẮT vi DANH MỤC BẢNG ix DANH MỤC HÌNH x MỞ ĐẦU Chương 1: CẢM BIẾN ADN ĐIỆN HÓA 1.1 Giới thiệu ADN 1.2 Cơ sở lý thuyết phương pháp điện hóa sử dụng nghiên cứu cảm biến ADN 1.2.1 Phương pháp quét vòng 1.2.2 Phương pháp quét xung vi phân (DPV) 11 1.2.3 Phương pháp qt sóng vng (SWV) 12 1.2.4 Phương pháp phổ tổng trở điện hoá (EIS) 13 1.3 Các phương pháp cố định chuỗi ssADN 15 1.3.1 Hấp phụ vật lý 15 1.3.2 Cố định liên kết cộng hóa trị 16 1.3.3 Cố định thông qua tương tác Avidin/Streptavidin -Biotin 18 1.3.4 Cố định phương pháp điện hóa 19 1.4 Phương pháp phát lai hóa ADN 21 1.4.1 Phương pháp đánh dấu 21 1.4.2 Phương pháp không đánh dấu 25 1.5 Ứng dụng cảm biến sinh học 29 Kết luận chương 29 Chương 2: NGHIÊN CỨU TỔNG HỢP VẬT LIỆU THANH NANO CeO2 & NANO COMPOSIT CeO2 NR@Ppy 31 2.1 Đặt vấn đề 31 2 Thực nghiệm 34 2.2.1 Hóa chất, thiết bị 34 2.2.2 Tổng hợp vật liệu nano CeO2 35 iii 2.2.3 Tổng hợp vật liệu nano composit CeO2 NR@Ppy 36 2.3 Kết thảo luận 37 2.3.1 Kết tổng hợp vật liệu nano CeO2 37 2.3.2 Kết tổng hợp vật liệu nano composit CeO2 NR@Ppy 47 Kết luận chương 53 Chương 3: PHÁT TRIỂN CẢM BIẾN ADN TRÊN CƠ SỞ CÁC THANH NANO CeO2 54 3.1 Đặt vấn đề 54 Thực nghiệm 57 3.2.1 Hóa chất 57 3.2.2 Cố định ssADN 58 3.2.3 Các phép đo điện hóa 60 3.3 Kết thảo luận 61 3.3.1 Kết cố định chuỗi ssADN 61 3.3.2 Đặc trưng cảm biến ADN 64 3.3.3 Tối ưu hóa điều kiện thực nghiệm 68 3.3.4 Độ lặp lại, độ ổn định, tính chọn lọc khả tái sử dụng 72 Kết luận chương 73 Chương 4: CẢM BIẾN SINH HỌC ADN TRÊN CƠ SỞ VẬT LIỆU NANO COMPOSIT CeO2 NR@Ppy 75 4.1 Đặt vấn đề 75 4.2 Thí nghiệm 76 4.2.1 Hóa chất 76 4.2.2 Cố định chuỗi ssADN dò lên bề mặt điện cực 77 4.2.3 Phân tích điện hóa 78 4.3 Kết thảo luận 78 4.3.1 Kết cố định ssADN 78 4.3.2 Đặc trưng điện hóa cảm biến sinh học ADN 80 4.3.3 Nghiên cứu ảnh hưởng số thơng số đến tín hiệu cảm biến 83 4.3.4 Độ chọn lọc, độ lặp lại độ ổn định cảm biến sinh học ADN 85 iv 4.3.5 Xác định vi khuẩn Salmonella phương pháp PCR so sánh với số liệu phát cảm biến ADN 87 Kết luận chương 88 KẾT LUẬN 89 KIẾN NGHỊ 90 DANH MỤC CÁC CƠNG TRÌNH ĐÃ CƠNG BỐ CỦA LUẬN ÁN 91 TÀI LIỆU THAM KHẢO 92 v DANH MỤC CÁC KÝ HIỆU VÀ CHỮ VIẾT TẮT Ký hiệu, TT chữ viết Tên Tiếng Anh Tên Tiếng Việt tắt ADN Acid deoxyribonucleic A xít deoxyribonucleic APTES 3-Aminopropyl-triethoxy- 3-Aminopropyl-triethoxy-silan silane ASV Adsortive Stripping Vơn-ampe hịa tan hấp phụ Voltammetry BSA Bovine Serum Albumin Bovine Serum Albumin CE Counter electrode Điện cực đối CPs Conducting Polyme(s) (Các) Polyme dẫn điện CTAB Cetyltrimethyl ammonium cetyltrimetyl amoni bromit bromide CV Cyclic Voltammetry Vơn - ampe vịng DAO Diamin Oxidase Enzym DAO 10 DIG Digoxigenin Digoxigenin 11 DPV Differential Pulse Quét xung vi sai Voltammetry 12 dsADN Double strand ADN Chuỗi ADN kép 13 EDC 1-Ethyl-3-3-Dimethyl- 1-Etyl-3-3-Dimetyl- aminopropyl Carbodiimide aminopropyl Cacbodiimit 14 EDS Energy dispersive X-ray Phổ tán sắc lượng tia X spectroscopy 15 EDTA Ethylene Diamine Tetracetic Etylen Diamin Tetracetic A xít Acid 16 EIS Electrochemical Impedance Phổ tổng trở điện hóa Spectrocopy 17 ELISA Enzym-linked Xét nghiệm miễn dịch liên kết Immunosorbent assay với enzym vi 18 FESEM Field Scaning Hiển vi điện tử quét phát xạ Emission trường Electron Microscopy 19 FTIR Fourier Transform Infrared Phổ Spectrocopy 20 hCG hồng ngoại biến đổi Fourier Human Chorionic hooc môn gonadotropin Gonadotropin 21 HRP Horseradish peroxide Horseradish Perô xít 22 IEP Isoelectric Point Điểm đẳng điện 23 ISFET Ion-Sensitive Field-Effect Trasistor hiệu ứng trường nhạy Transistor ion 24 ITO Indium Tin Oxide Ơ xít thiếc indi 25 LOD Limit of Detection Giới hạn phát 26 LOQ Limit of Quantification Giới hạn định lượng 27 MIA 1-Methylimidazole 1-Metyl imidazol 28 NRs Nanorods Các nano 29 PABA Acid Para-Aminobenzoic A xít 4-Aminobenzoic 30 PANAM Poly Amidoamine Poly amidoamin 31 PANi Polyaniline Polyanilin 32 PBS Phosphat Buffer Saline Đệm phốt phát 33 PCR Polymerase Chain Reaction Phản ứng chuỗi Polyme 34 Ppy Polypyrrole Poly pyrol 35 Py Pyrrole Pyrol 36 RE Reference electrode Điện cực so sánh 37 SCE Saturated Calomel Electrode Điện cực Calomen bão hòa 38 SMOs Solid Metal Oxides xít kim loại bán dẫn 39 ssADN Single strand AND Chuỗi ADN đơn 40 SWV Square Wave Voltammetry Qt sóng vng 41 TEM Transmission electron Hiển vi điện tử truyền qua microscopy 42 TGA Thermogravimetric Analysis vii Phân tích nhiệt 43 TMC N-trimetyl chitosan N-trimetyl chitosan 44 WE Working electrode Điện cực làm việc 45 XRD X-ray Diffraction Nhiễu xạ tia X viii DANH MỤC CÁC CƠNG TRÌNH ĐÃ CƠNG BỐ CỦA LUẬN ÁN A Tạp chí quốc tế Nguyen Thi Nguyet, Le Thi Hai Yen, Pham Hung Vuong, Phuong Dinh Tam, Tran Trung (2018), 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NR@Ppy Các kỹ thuật sử dụng để tổng hợp vật liệu nano thường áp dụng để tổng hợp vật liệu có cấu trúc lõi vỏ Vật liệu lõi vỏ tổng hợp từ tiền... CeO2 vật liệu nano composit CeO2 NR@Ppy có cấu trúc lõi vỏ  Phát triển cảm biến ADN hiệu suất cao sở vật liệu nano chiều CeO2 chế tạo Nội dung nghiên cứu luận án - Nghiên cứu tổng hợp vật liệu nano