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Published on: June 19, 2015
Distribution and Molecular Characterization of Functional Class 2 Integrons in Clinical Proteus mirabilis Isolates
Wenjun Lu1, Quedan Qiu2, Keda Chen2
1Intensive Care Units of Ningbo Medical Centre Lihuili Hospital, Ningbo University, Ningbo, Zhejiang Province, People's Republic of China.
Background:
Integrons are the main mode of horizontal transmission of drug-resistance genes and are closely related to drug resistance in clinical bacteria. In this study, the distributions of class 1, 2, and 3 integron gene cassettes were investigated in 150 Proteus mirabilis (P. mirabilis) isolates from patients, and molecular characterization of functional class 2 integrons was further analyzed.
Methods:
Class 1, 2, and 3 integrons were screened by polymerase chain reaction (PCR) in 150 clinical P. mirabilis isolates. The variable regions of the integrons were determined by restriction analysis and sequencing. Internal stop codons mutations in class 2 integrons and their common promoters were also determined by sequencing. Enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) was used to analyze the phylogenetic relations of class 2 integron-positive isolates.
Results:
Class 1 integrons were detected in 69 (46%) of 150 P. mirabilis isolates, and six different gene cassette arrays were detected, with the most prevalent being dfrA32-aadA2. Class 2 integrons were detected in 61 (40.7%) of 150 P. mirabilis isolates, and three different gene cassette arrays were detected, including sat2-aadA1, which was detected for the first time in a class 2 integron. Nearly similar ERIC-PCR fingerprinting patterns were detected in 45 (73.8%) of 61 class 2 integron-positive isolates. The functional class 2 integron was detected in three P. mirabilis isolates having the same gene cassette, dfrA1-sat2-aadA1, in the variable region and four novel open reading frames with unknown functions. Same PintI2 and Pc promoters were detected in these three functional class 2 integron isolates, as was found in other class 2 integron isolates. However, these three strains did not totally show identical homology and drug sensitivity.
Conclusion:
Although functional class 2 integrons have low distribution and relatively conserved molecular characteristics, they can still form clinical dissemination and drug resistance expression.
Insights
Integrons are key to spreading drug resistance in bacteria. This study found class 1 and 2 integrons in Proteus mirabilis, with class 2 integrons showing conserved features despite low distribution, contributing to drug resistance.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Integrons facilitate horizontal gene transfer of drug resistance genes, impacting clinical bacteria.
- Proteus mirabilis is a significant clinical pathogen where integrons play a role in antimicrobial resistance.
Purpose of the Study:
- To investigate the distribution of class 1, 2, and 3 integron gene cassettes in clinical Proteus mirabilis isolates.
- To perform molecular characterization of functional class 2 integrons and analyze their phylogenetic relationships.
Main Methods:
- Screening of 150 P. mirabilis isolates for class 1, 2, and 3 integrons using Polymerase Chain Reaction (PCR).
- Analysis of integron variable regions via restriction analysis and sequencing.
- Sequencing of internal stop codons and promoters in class 2 integrons.
- Phylogenetic analysis of class 2 integron-positive isolates using Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR).
Main Results:
- Class 1 integrons found in 46% of isolates, with dfrA32-aadA2 being the most common array.
- Class 2 integrons detected in 40.7% of isolates, revealing the sat2-aadA1 array for the first time in this class.
- Functional class 2 integrons with conserved P(intI2) and Pc promoters were identified in three isolates, alongside novel open reading frames.
Conclusions:
- Functional class 2 integrons, despite limited distribution and conserved features, contribute to the clinical spread and expression of drug resistance.
- The study highlights the role of integrons in antimicrobial resistance within Proteus mirabilis clinical isolates.
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