Xây dựng quy trình phát hiện nhanh ngô chứa gen Cry1Ab dựa trên kỹ thuật LAMP

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Tác giả: Viết Thế Hồ, Ngọc Giàu Lê, Thị Kim Anh Ngô, Thị An Nguyễn, Thị Hương Nguyễn, Thị Mỹ Hạnh Trần

Ngôn ngữ: vie

Ký hiệu phân loại: 641.3315 Food

Thông tin xuất bản: Tạp chí khoa học công nghệ và thực phẩm - Đại học Công nghiệp Thực phẩm TP Hồ Chí Minh, 2020

Mô tả vật lý: 37-45

Bộ sưu tập: Metadata

ID: 492405

Thêm vào giỏ

Việt Nam đã cho phép trồng ngô biến đổi gen có chứa gen Cry1Ab để tăng khả năng kháng sâu đục thân. Tuy nhiên, vẫn còn nhiều ý kiến tranh cãi về mức độ an toàn của loại cây trồng mới này, vì vậy đã có nhiều phương pháp sử dụng để phát hiện sự hiện diện của gen Cry1Ab trong cây ngô. Các phương pháp hiện tại phần lớn đều phụ thuộc nhiều vào trang thiết bị phòng thí nghiệm và khó thực hiện ngoài thực địa. Gần đây, kỹ thuật khuếch đại DNA đẳng nhiệt LAMP (Loop Mediated Isothermal Amplification) đã được sử dụng để phát hiện nhanh các đối tượng gây bệnh trên người và vật nuôi một cách nhanh chóng, với độ chính xác cao và ít yêu cầu về trang thiết bị đắt tiền. Trong nghiên cứu này, quy trình phát hiện ngô biến đổi gen bằng kỹ thuật LAMP đã được xây dựng thành công thông qua khảo sát các yếu tố ảnh hưởng tới phản ứng. Kết quả cho thấy nồng độ phù hợp của các mồi FIP/BIP và F3/B3 là 0,4 μM với nhiệt độ phản ứng phù hợp ở 60 °C. Ngoài ra, qua khảo sát về ngưỡng phát hiện của phương pháp LAMP so với phương pháp Realtime PCR cũng cho thấy độ nhạy của phản ứng LAMP cao hơn phản ứng Realtime PCR. Ngoài ra, khi sử dụng thuốc nhuộm HNB ở nồng độ 80 μM, có thể phân biệt được giữa ngô không chuyển gen và ngô chứa gen Cry1Ab. Kết quả này là tiền đề để thiết kế KIT phát hiện ngô biến đổi gen ngoài thực địa.Vietnam has allowed the cultivation of gentically modified maize containing Cry1Ab gene to increase resistance to stem borer. Despite many advantages, there are still many debates about the safety of this crop so there are many methods used to detect them. However, the current methods rely heavily on laboratory equipment and are difficult to perform in the field. Recently, the LAMP (Loop Mediated Isothermal Amplification) DNA amplification technique has been used to quickly detect human and animal disease objects, with high accuracy and less requirements for expensive equipment. Within the scope of this study, we have successfully developed a process to detect gentically modified maize using LAMP technology. The results of the study have optimized parameters for LAMP reaction including concentration of FIP/BIP and F3/B3 of 0.4 μM with primer pairing temperature at 60 °C. The authors also found that the sensitivity of LAMP reaction was better than that of Realtime PCR in studied range. When using HNB dye at a concentration of 80 nM, it was possible to distinguish between the non-transgenic and Cry1ab gene containing maize. This result is a potential for designing KIT to detect gentically modified corn in the field.

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