期刊
MOLECULAR BIOLOGY REPORTS
卷 47, 期 10, 页码 7341-7348出版社
SPRINGER
DOI: 10.1007/s11033-020-05789-6
关键词
2-D multiplex qPCR; Melting temperature (Tm); Fluorescence probe; Antibiotic-resistance gene; Amplicon; Melting curve
资金
- National Science and Technology Major Project of China [2017ZX10103009-002]
- One Belt One Road project of the Chinese Academy of Sciences [153831KYSB20170043]
- 133 projects of Institut Pasteur of Shanghai, CAS
Simple, multiplex qPCR methods are advantages for rapid molecular diagnosis of multiple antibiotics-resistant genes simultaneously. However, the number of genes can be detected in a single reaction tube is often limited by the fluorescence channels of a real-time PCR instrument. In this study, we developed a simple 2-D multiplex qPCR method by combining the probe colors and amplicon Tm values to overcome the mechanical limit of the machine. The principle of the novel assay was validated by detection of nine bacterial antibiotic-resistance genes (KPC, NDM, VIM, OXA-48, GES, CIT, EBC, ACC and DHA) in a single reaction tube. This assay is highly sensitive within a range of 30-3000 copies per reaction. The simplicity, rapidity, high sensitivity and specificity, and low cost of the novel method make it a promising tool for developing clinical diagnostic kits for monitoring resistance and other genetic determinants of infectious diseases.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据