Gene expression of katB and oxyR using qRT-PCR in multidrug-resistant Pseudomonas aeruginosa isolates from burn and wound patients | ||
| Microbes and Infectious Diseases | ||
| Articles in Press, Accepted Manuscript, Available Online from 15 October 2025 | ||
| Document Type: Original Article | ||
| DOI: 10.21608/mid.2025.425184.3237 | ||
| Authors | ||
| Ziyad Kadhem Dahil Alburki* 1; Samira Gjir Jremich2 | ||
| 1Department of Biology, College of Education , University of AL-Qadyssia, Iraq | ||
| 2Department of Pathological Analysis, College of Science, University of AL-Qadyssia, Iraq | ||
| Abstract | ||
| Background: Pseudomonas aeruginosa is one of the most common hospital-acquired pathogens, particularly in burn units and intensive care units, because it can establish acute or chronic infections that can be deadly. Aims of study: This current study aimed to examine peroxidase gene expression in MDR P. burn isolates from burn and wound patients, and using quantitative (qPCR) to elucidate expression patterns and correlate them to resistance profiles. Methods: Nine isolates were selected; eight were treated with hydrogen peroxide (0.5 mM H₂O₂) for the induction of oxidative stress, one untreated isolate served as control. Cultures were incubated in liquid medium at 37 °C with shaking until the logarithmic phase. The cells were harvested by centrifugation, and the harvested cells were stored at –70°C for RNA extraction. Results: the results showed that the amplification curves of the katB and oxyR genes in Pseudomonas aeruginosa isolates exposed to 0.5 mM hydrogen peroxide. The Ct values varied amongst isolates suggesting variation in the genetic response. The melting curve of the qRT-PCR products for all the qRT-PCR runs without any additional peaks we had a single sharp peak at Tm = 84.38°C. katB expression increased up to 2.6-fold under oxidative stress. The data shows high specificity of the amplification and rules out nonspecific products and primer-dimers. Therefore, additional evidence supporting credibility of the data has been provided. Conclusions: PCR amplification of a species-specific gene was performed for 11 of the 30 isolates. The oxidative stress response of nine isolates following exposure to 0.5 mM H₂O₂ was assessed using qRT-PCR analysis. isolates 2,3, and 5 exhibited low induction capacity for katB genes (≤1.6 fold) and approximately baseline expression for oxyR genes. | ||
| Keywords | ||
| P. aeruginosa; (MDR); PCR (qPCR); burns and wound | ||
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