ĐÁNH GIÁ MỘT SỐ SINH PHẨM VÀ PHƯƠNG PHÁP TÁCH CHIẾT ADN VI KHUẨN MYCOBACTERIA TRÊN MẪU BỆNH PHẨM THU THẬP TỪ BỆNH NHÂN NHIỄM LAO
Trung tâm Nhiệt đới Việt - Nga
Số 63, Nguyễn Văn Huyên - Nghĩa Đô - Cầu Giấy - Hà Nội
Số điện thoại: 0963122607; Email: leanhbio@gmail.com
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Tóm tắt
EVALUATION SOME DNA EXTRACTION KITS AND METHODS FOR MYCOBACTERIA FROM SPUTUMS OF TUBERCULOSIS INFECTED CASES
Tuberculosis is still burden in developing countries, including Viet Nam. Molecular techniques such as PCR, Real-time PCR play an important role in tuberculosis diagnostics. Specimens for tuberculosis testing are complex such as sputums, gastric fluids. DNA extraction are commonly performed by commercial kits, with little study about efficacy for tuberculosis testing assays. This study describes efficacy of three kits G-Spin (Intron-Korea), RIBO- sorb (Amplisens-Russia), DNA/RNA Prep (Sacace-Italy), and Salting-out method using basic reagents. Samples were 15 sputums collected from tuberculosis infected cases. As the result, recovered ADN was highest in DNA/RNA Prep, folowed by Salting-out, G-Spin, and last by RIBO - sorb. Oder of purification value was RIBO - sorb, G-Spin, Salting-out, and DNA/RNA Prep. About PCR for 16S gene, 15/15 samples were succeed in G-Spin and DNA/RNA Prep, 14/15 from RIBO-prep, 13/15 from Salting-out. Real-time PCR Mycobacterium tuberculosis (Sacace-Italy) detected 14/15 samples from G-Spin, DNA/RNA Prep, and RIBO-sorb, but Salting-out detected 13/15 samples. Therefore, G-Spin and DNA/RNA Prep show higher efficacy in amplication of 16S gene and Real-time PCR assays for tuberculosis. RIBO- sorb has equal efficacy in Realtime PCR, but more priority in purification. Salting-out perform equally in recovery and purification with commercial kits, but limited in efficacy of Mycobacteria genome amplification.
Từ khóa
DNA extraction, Mycobacteria, Salting-out, 16S, Realtime PCR, tách chiết ADN
Chi tiết bài viết
Tài liệu tham khảo
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