Establishment of loop-mediated isothermal amplification technique for rapid detection of NDM-1 gene.
- Author:
Yuanyi ZHANG
1
;
Na WU
;
Baoli ZHU
;
Lei CHEN
;
Yuzhuo ZHU
Author Information
1. Laboratory of Medical Genetics, Jilin University, Changchun 130021, China.
- Publication Type:Journal Article
- MeSH:
Bacteria;
enzymology;
genetics;
Bacteriological Techniques;
methods;
DNA, Bacterial;
genetics;
Escherichia coli;
enzymology;
genetics;
Klebsiella pneumoniae;
enzymology;
genetics;
Nucleic Acid Amplification Techniques;
methods;
Sensitivity and Specificity;
Staphylococcus aureus;
enzymology;
genetics;
Streptococcus pneumoniae;
enzymology;
genetics;
beta-Lactamases;
genetics
- From:
Chinese Journal of Biotechnology
2011;27(8):1232-1238
- CountryChina
- Language:Chinese
-
Abstract:
We established a rapid detection method of New Delhi Metallo-beta-Lactamase Gene (NDM-1) based on Loop-mediated Isothermal Amplification (LAMP). With the application of LAMP, we designed four sets of LAMP premiers, using NDM-1 gene as the target sequence, and selected the set of optimal primers. Meanwhile, we established optimal reaction systems and conditions to carry out the sensitivity and specificity experiments. The experiment results showed that the whole detection process took only one hour and could be observed visually. In the experiment of sensitivity, NDM-1 gene had a detection limit of 6 copies in each reaction. In the experiment of specificity, we detected NDM-1 gene in 4 pathogen strains (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae), and the total DNA from intestinal microbes and the total DNA from soil microbes. We had not detected the amplification reactions. The detection method established could rapidly detect NDM-1 gene and visualize the experiment result. The method is easy to operate and has high sensitivity and specificity and thus has great application value in basic research laboratories, emergent detection and spot detection.
