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High-throughput Assay to Phenotype Salmonella enterica Typhimurium Association, Invasion, and Replication in Macrophages
Published on: August 11, 2014
Integron-Associated Antibiotic Resistance in Salmonella typhi
Z Mohammed Jassim1, F Mohammed Obead2, S Neama2
1College of biotechnology, Al-Qasim Green University, Al Qasim, Iraq.
Abstract:
Salmonella enteric serovar Typhi (S. typhi) and paratyphi (S. paratyphi) bacteria exclusively found in humans, cause typhoid fever, an acute, and possibly deadly systemic infection. Typhoid fever is caused by a species of rod-shaped, Gram-negative Enterobacteriaceae called S. typhi. The present study aimed to examine the intI gene and investigate the possible relation between this gene and multi-drug resistance in S. typhi. A total of 30 blood samples were obtained from patients who were suspicious of typhoid fever using the direct strategy of inoculation. Each specimen was injected into a culture of a selective medium, such as XLD and SS agar, and then incubated at 37°C for 24 h. The genomic DNA was extracted through a boiling process. Tris-EDTA was used to suspend bacterial colonies cultured on MacConkey agar plates. The suspension of bacterial colonies was centrifuged for 5 min at 8000×g and for 20 min at -20°C which lyses the organisms and extracts the DNA from the buffer. The supernatant is then transferred to a fresh Eppendorf tube. Gel electrophoresis was carried out utilizing a UV transilluminator. The intI gene for S. typhi was found using a PCR test. The antibiotic sensitivity testing showed that the S. typhi isolates were classed as multi-resistant. These results were confirmed using the polymerase chain reaction (PCR) technique using intI gene where twenty specimens isolated from typhoid patients were positive for S. typhi.
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