Extensive Expression of the Virulome Related to Antibiotic Genotyping in Nosocomial Strains of <i>Klebsiella pneumoniae</i>
The emergence of hyper-virulent and multidrug-resistant (MDR) strains of <i>Klebsiella pneumoniae</i> isolated from patients with hospital- and community-acquired infections is a serious health problem that increases mortality. The molecular analysis of virulome expression related to ant...
Main Authors: | , , , , , , , , , |
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Format: | Article |
Language: | English |
Published: |
MDPI AG
2023-09-01
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Series: | International Journal of Molecular Sciences |
Subjects: | |
Online Access: | https://www.mdpi.com/1422-0067/24/19/14754 |
Summary: | The emergence of hyper-virulent and multidrug-resistant (MDR) strains of <i>Klebsiella pneumoniae</i> isolated from patients with hospital- and community-acquired infections is a serious health problem that increases mortality. The molecular analysis of virulome expression related to antimicrobial-resistant genotype and infection type in <i>K. pneumoniae</i> strains isolated from patients with hospital- and community-acquired infections has been poorly studied. In this study, we analyzed the overall expression of the virulence genotype associated with the antimicrobial resistance genotype and pulse field gel electrophoresis (PFGE) type (PFtype) in <i>K. pneumoniae</i>. We studied 25 strains of <i>K. pneumoniae</i> isolated from patients who developed bacteremia and pneumonia during their hospital stay and 125 strains from outpatients who acquired community-acquired infections. Susceptibility to 12 antimicrobials was determined by Kirby–Bauer. The identification of <i>K. pneumoniae</i> and antibiotic-resistance genes was performed using polymerase chain reaction (PCR). To promote the expression of the virulence genes of <i>K. pneumoniae</i>, an in vitro infection model was used in human epithelial cell lines A549 and A431. Bacterial RNA was extracted with the QIAcube robotic workstation, and reverse transcription to cDNA was performed with the Reverse Transcription QuantiTect kit (Qiagen). The determination of the expression of the virulence genes was performed by real-time PCR. In addition, 57.3% (n = 86) of the strains isolated from patients with hospital- and community-acquired infections were multidrug-resistant (MDR), mainly to beta-lactam antibiotics (CB, AM, CFX, and CF), aminoglycosides (GE), quinolones (CPF and NOF), nitrofurantoin (NF), and sulfamethoxazole/trimethoprim (SXT). The most frequently expressed genes among strains isolated from hospital- and community-acquired infections were adhesion-type, <i>ycfm</i> (80%), <i>mrkD</i> (51.3%), and <i>fimH</i> (30.7%); iron uptake, <i>irp2</i> (84%)<i>, fyuA</i> (68.7%), <i>entB</i> (64.7%), and <i>irp1</i> (56.7%); and protectins, <i>rpmA</i> (26%), which were related to antibiotic-resistance genes, <i>bla<sub>TEM</sub></i> (96%), <i>bla<sub>SHV</sub></i> (64%), <i>bla<sub>CITM</sub></i> (52.6%), <i>bla<sub>CTXM-1</sub></i> (44.7%), <i>tetA</i> (74%), <i>sul1</i> (57.3%), <i>aac(3)-IV</i> (40.7%), and <i>aadA1</i> (36%). The results showed the existence of different patterns of expression of virulome related to the genotype of resistance to antimicrobials and to the PFtypes in the strains of <i>K. pneumoniae</i> that cause hospital- and community-acquired infections. These findings are important and may contribute to improving medical treatment strategies against infections caused by <i>K. pneumoniae</i>. |
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ISSN: | 1661-6596 1422-0067 |