Quy trình duplex pcr phát hiện đồng thời Vibrio parahaemolyticus và white spot syndrome virus (wssv) gây bệnh trên tôm biển

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Tác giả: Mộng Huyền Hồng, Thị Kim Ngọc Lê, Quí Nhân Nguyễn, Quốc Khánh Nguyễn, Minh Nhựt Phạm, Thị Tuyết Hoa Trần

Ngôn ngữ: vie

Ký hiệu phân loại: 664.94 Fishes and shellfish

Thông tin xuất bản: Khoa học (Đại học Hạ Long), 2022

Mô tả vật lý: 89-109

Bộ sưu tập: Metadata

ID: 500823

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The intensification of farming areas and climate change have made the shrimp farming industry suffer from various obstacles, especially the loss of productivity due to pathogens. White Spot Syndrome Virus(WSSV) and Vibrio parahaemolyticus(VpAHPND) were reported to cause huge economic losses to marine shrimp farmers. When causing diseases or outbreaks, they are extremely hard to be controlled. Therefore, detecting and screening out the pathogens are crucial. This study was conducted to optimize a duplex PCR (dPCR) protocol, which is able to detect WSSV and VpAHPND simultaneously. Benefits of the protocol are noticeable, such as resulting fast, and reducing the concentration of primer, dNTPs, and MgCl2 while providing substantial sensitivity and specificity. The results showed that (i) The PCR protocol was successfully optimized to detect WSSV and VpAHPND simultaneously and its specificity was determined. (ii) For the DNA template of WSSV and V. parahaemolyticus, the protocol required 100 pg/µL and 10 ng/µl, respectively. (iii) The duplex PCR was successfully applied to detect WSSV and V. parahaemolyticus in marine shrimp samples.Với việc tăng cường diện tích nuôi cùng với biến đổi khí hậu, ngành nuôi tôm hiện đang gặp nhiều trở ngại, đặc biệt là thiệt hại do mầm bệnh. Sự cần thiết phải phát hiện và sàng lọc các mầm bệnh là một trong những nhiệm vụ quan trọng nhất vì rất khó để kiểm soát khi nào dịch bùng phát. Nghiên cứu này được thực hiện nhằm tối ưu hóa quy trình duplex PCR (dPCR) phát hiện đồng thời vi-rút gây bệnh đốm trắng (WSSV) và Vibrio parahaemolyticus, hai mầm bệnh gây thiệt hại lớn về mặt kinh tế cho ngành nuôi tôm biển. Mục tiêu chính của nghiên cứu này là chuẩn hóa giảm nồng độ mồi, dNTPs và MgCl2 nhằm phát triển quy trình có độ nhạy và độ đặc hiệu cao và khả năng phát hiện thời gian ngắn. Kết quả cho thấy: (i) Phương pháp PCR được tối ưu hóa phát hiện đồng thời vi-rút gây bệnh đốm trắng (WSSV) và Vibrio parahaemolyticus (VpAHPND)
(ii) Cụ thể của PCR tối ưu hóa là 100 pg/µL đối với WSSV và 10 ng/µl đối với V. parahaemolyticus
(iii) Xác định tính đặc hiệu và ứng dụng của PCR duplex để phát hiện đồng thời vi-rút gây bệnh đốm trắng (WSSV) và V. parahaemolyticus (VpAHPND) trên tôm biển.

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