NGHIÊN CỨU TẠO ARMORED RNA LÀM CHỨNG DƯƠNG CHO REAL-TIME RT-PCR PHÁT HIỆN VIRUS MARBURG

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Tác giả: Liêm Chính Bùi, Quang Hòa Lê, Minh Anh Nguyễn, Thị Nga Nguyễn, Thị Thu Hoài Nguyễn, Thị Thùy Linh Nguyễn, Thị Thu Hương Phạm, Huyền Nhung Phùng, Khánh Ngọc Trần, Thanh Hương Vương

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Phân tích Hóa, Lý và Sinh học, 2022

Mô tả vật lý: tr.158

Bộ sưu tập: Metadata

ID: 337848

Marburg virus is the causative agent of a severe viral hemorrhagic fever in human with high mortality rate. This study aims to prepare virus-modified particles, based on MS2 bacteriophage, to be used as a positive control for the diagnosis of Marburg virus by Real-time RT-PCR. To this aim, the target region of Real-time RT-PCR assay was cloned into the pMC037-HisMS2_PLP_pac plasmid, generating expression vector pMS2-MARV-NP. MS2-like particles were then produced in E. coli BL21(DE3) and were purified by Ni-NTA affinity chromatography. Analysis by SDS-PAGE and agarose gel electrophoresis, transmission electron microscopy, and real-time RT-PCR showed that the His-tagged Armored RNA particles carrying the target region were successfully assembled and purified at high concentration (3.9 × 108 copies/µL by RT-qPCR quantification) while DNA plasmid contamination was negligible. Most importantly, these armored RNA particles were found to be resistant to RNase A. These findings reveal the potential of Armored RNA technology for rapid preparation of ribonucleaseresistant viral RNA controls.
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