TỐI ƯU HÓA QUY TRÌNH PHÁT HIỆN VI KHUẨN Salmonella spp. GÂY BỆNH TRÊN THỰC PHẨM BẰNG KỸ THUẬT LAMP (LOOP-MEDIATED ISOTHERMAL AMPLIFICATION)

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Tác giả: Thị Quỳnh Hương Đoàn, Phạm Trúc Phương Nguyễn, Văn Lượng Nguyễn

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học và Công nghệ - Đại học Thái Nguyên, 2022

Mô tả vật lý: tr.92-101

Bộ sưu tập: Metadata

ID: 332078

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Method for early and rapid detection of foodborne pathogens caused by Salmonella bacteria will help control contaminated food in the market. Therefore, in this study, a procedure for detecting Salmonella in foods by LAMP technique has been developed to quickly and accurately detect strains of Salmonella spp. based on the invA2 primer set specifically for the invA gene region of Salmonella. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity. The optimized LAMP protocol include concentrations of primers F3B3/FIPBIP 200Nm/1600nM (1:8) and Betaine 0,8M, 60 minutes heating at 63oC, the specificity of LAMP protocol for the detection of Salmonella spp. was 100% while sensitivity was 5pg/reaction (on DNA template), 4.1x101CFU/ml (on bacterial strain). The results were compared with those obtained by realtime PCR.Việc phát hiện sớm và nhanh các mầm bệnh trên thực phẩm do vi khuẩn Salmonella gây ra sẽ giúp kiểm soát được thực phẩm bẩn trên thị trường. Vì vậy, trong nghiên cứu này đã xây dựng quy trình phát hiện Salmonella trong các loại thực phẩm bằng kỹ thuật LAMP để phát hiện nhanh và chính xác các chủng vi khuẩn Salmonella spp. dựa trên bộ mồi invA2 chuyên biệt cho vùng gen invA của vi khuẩn Salmonella. Sau quá trình tối ưu các thành phần và điều kiện cần thiết như tỉ lệ mồi F3B3/FIPBIP 200Nm/1600nM (1:8), nồng độ Betaine 0,8M, nhiệt độ 63oC và thời gian là 45 phút, quy trình LAMP phát hiện Salmonella spp. có độ đặc hiệu kỹ thuật 100%, độ nhạy kỹ thuật trên nồng độ DNA là 5pg/phản ứng
độ nhạy kỹ thuật của LAMP trên chủng khuẩn là 4,1x101CFU/ml. Độ nhạy của xét nghiệm LAMP được so sánh với phản ứng realtime PCR.

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