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Updated: Feb 18, 2026
Screening Foodstuffs for Class 1 Integrons and Gene Cassettes
Published on: June 19, 2015
Virulence and resistance on various pathogens mediated by mobile genetic integrons via high flux assays
Yukui Zhong1, Huamin Zhong1, Qiulian Deng1
1Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou 510120, China.
Background:
Recognized as a resistance mechanism responsible for the emergence and prevalence of antimicrobial resistance, integron is widely distributed and spread among clinical microorganisms and play a key role in the dissemination of such antimicrobial resistance, which may eventually contribute to the unleashing of "Super Bugs" In this study, detection assays based on loop-mediated isothermal amplification (LAMP) methodologies targeting on class 1 to class 3 integrase genes was developed and evaluated.
Methods:
LAMP methodology was employed to develop novel detection assays on class 1, 2 and 3 integrons. Firstly, this protocol was specifically designed to detect such integrons by targeting integrase genes intI1, intI2 and intI3. Development, evaluation and optimization of such LAMP assays was studied, including the reaction temperature, volumn, time, sensitivity and specificity of both primers and targets. A total of 1082 strains, including 397 integron positive and 685 integron negative microorganisms, were included for the application verification of the established LAMP assays.
Results:
The indispensability of each primer was confirmed, and the optimal amplification was obtained under 63 °C for 45 min, with 25 μl reaction found to be the most cost-efficient volume. As application was concerned, all of the 397 integron-positive isolates yielded positive amplicons and other 685 integron-negative bacteria were negative for the integron-LAMP assays, revealing totaling 100% detection rate and specificity.
Conclusions:
The established integron-LAMP assays was demonstrated to be a valid and rapid detection method for integrons screening, which may aid in both the laboratory and clinical integron screening for microorganisms.
Insights
New loop-mediated isothermal amplification (LAMP) assays rapidly detect integrons, crucial for antimicrobial resistance. This method offers 100% accuracy for screening integron-positive and negative microorganisms in clinical settings.
Area of Science:
- Microbiology
- Molecular Biology
- Antimicrobial Resistance Research
Background:
- Integrons are key genetic elements driving antimicrobial resistance (AMR) and the spread of
Purpose of the Study:
- Develop and validate novel loop-mediated isothermal amplification (LAMP) assays for detecting class 1, 2, and 3 integrons.
- Establish a rapid and accurate method for identifying integron-carrying microorganisms.
Main Methods:
- Loop-mediated isothermal amplification (LAMP) methodology was optimized to target integrase genes (intI1, intI2, intI3) of integrons.
- Assay parameters including temperature, time, volume, sensitivity, and specificity were evaluated.
- The developed LAMP assays were tested on 1082 bacterial strains (397 integron-positive, 685 integron-negative).
Main Results:
- Optimal LAMP conditions determined as 63°C for 45 minutes with a 25 μl reaction volume.
- The assays demonstrated 100% detection rate and specificity, correctly identifying all 397 integron-positive and 685 integron-negative isolates.
- All primers and targets were confirmed as indispensable for accurate amplification.
Conclusions:
- The developed integron-LAMP assays provide a valid, rapid, and highly accurate method for screening integrons.
- This technique can significantly aid in both laboratory and clinical settings for identifying microorganisms carrying integrons.
- The assays contribute to efforts in combating the spread of antimicrobial resistance.
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