A loop-mediated isothermal amplification method for rapid detection of NDM-1 gene
Jing Qi1, Yijun Du, Xiaoling Zhu
1Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, China.
Microbial Drug Resistance (Larchmont, N.Y.)
|March 16, 2012
Summary
A new loop-mediated isothermal amplification (LAMP) method rapidly and sensitively detects the New Delhi Metallo-β-lactamase-1 (NDM-1) gene. This accurate and visual assay is valuable for on-the-spot NDM-1 investigations.
Area of Science:
- Microbiology
- Molecular Biology
- Antimicrobial Resistance
Background:
- The New Delhi Metallo-β-lactamase-1 (NDM-1) gene confers resistance to carbapenems, a critical class of antibiotics.
- Rapid and accurate detection of NDM-1 is essential for infection control and surveillance.
- Existing methods like PCR can be time-consuming and require specialized equipment.
Purpose of the Study:
- To develop and evaluate a novel loop-mediated isothermal amplification (LAMP) method for NDM-1 detection.
- To assess the specificity, sensitivity, and speed of the developed LAMP assay.
- To compare the LAMP method with conventional polymerase chain reaction (PCR) for clinical sample analysis.
Main Methods:
- Design of six specific primers targeting eight sequences of the NDM-1 gene.
- Isothermal amplification reaction conducted at 65°C for 40 minutes.
- Evaluation of specificity and sensitivity using reference strains and clinical samples.
- Comparison of LAMP results with conventional PCR.
Main Results:
- The LAMP assay demonstrated high specificity and sensitivity, surpassing conventional PCR.
- The limit of detection was as low as 1 pg of genomic DNA for the NDM-1 gene.
- 100% concordance was observed between LAMP and PCR for 345 clinical samples.
- LAMP successfully detected NDM-1 in three samples contaminated and missed by PCR.
Conclusions:
- The developed LAMP method offers a rapid, visual, specific, accurate, and sensitive approach for NDM-1 gene detection.
- This assay is particularly suitable for on-the-spot investigations and resource-limited settings.
- LAMP presents a valuable alternative to PCR for timely NDM-1 surveillance and diagnostics.
