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Related Experiment Video

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Screening Foodstuffs for Class 1 Integrons and Gene Cassettes
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Two-dimensional PCR for detecting class 1, 2 and 3 integrons.

Wenwen Zhu1, Tong Wang2, Yu Zhu2

  • 1School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangdong Province, Guangzhou 510515, China.

Heliyon
|December 1, 2022
PubMed
Summary

A new two-dimensional PCR (2D-PCR) method efficiently detects three major integrons simultaneously. This rapid technique aids in large-scale screening of antibiotic resistance genes in clinical bacterial isolates.

Keywords:
2D-PCRAntibiotic resistanceGene cassetteIntegraseIntegron

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Integrons are crucial genetic elements facilitating the spread of antibiotic resistance genes via site-specific recombination.
  • Detecting and characterizing integrons is vital for understanding and combating bacterial antibiotic resistance.

Purpose of the Study:

  • To develop and validate a novel two-dimensional PCR (2D-PCR) method for simultaneous detection of three major integron classes (intI1, intI2, intI3).
  • To assess the sensitivity, specificity, and applicability of the 2D-PCR method in clinical settings.

Main Methods:

  • Development of a 2D-PCR assay utilizing base quenching probe technology for simultaneous integron detection.
  • Evaluation of minimum detection limits using plasmids with varying integron concentrations.
  • Assessment of specificity through screening 105 clinical *Proteus* isolates and comparison with traditional PCR.

Main Results:

  • The 2D-PCR method successfully distinguished between the three integron classes based on a ~10°C melting temperature difference.
  • Minimum detection limits for intI1, intI2, and intI3 were below 102 copies/μl.
  • Detection results from clinical isolates showed high concordance with traditional PCR methods.

Conclusions:

  • The developed 2D-PCR method is rapid, economical, and high-throughput for simultaneous detection and typing of three major integron classes.
  • This method is suitable for large-scale integron screening and typing in clinical bacterial isolates, aiding in antibiotic resistance surveillance.