Journal of Isfahan Medical School

Journal of Isfahan Medical School

Detection of Tetracycline-Resistant Genes by Multiplex Polymerase Chain Reaction in Clinical Isolates of Klebsiella Pneumonia

Document Type : Original Article (s)

Authors
Manar Naji-Hamad 1
Mahboobeh Nakhaei-Moghaddam 2
1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
2 Associate Professor, Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
10.22122/jims.v39i647.14366
Abstract
Background: Klebsiella pneumoniae (K. pneumonia) is an opportunistic pathogen and one of the most common causes of nosocomial infections, some of which are now resistant to antibiotics and make infection difficult to treat. The main purpose of this study was to use multiplex polymerase chain reaction (PCR) to detect tetracycline-resistance genes in clinical isolates of K. pneumonia.Methods: After collecting clinical samples from hospitals in Mashhad City, Iran, in 2021, K. pneumoniae was isolated and identified by biochemical and molecular methods (Kp16S rRNA specific primer). Antibiotic-resistance pattern was determined for 30 isolates by disk diffusion method. After designing primer and its blast, detection of tet genes was optimized by PCR. Finally, multiplex PCR was set up using three pairs of primers and positive control.Findings: The highest and lowest antibiotic resistance of the isolates was for ampicillin (90%) and ceftriaxone (26.66%), respectively, and 53.33% of Klebsiella isolates were phenotypically resistant to tetracycline. Multiplex PCR was set up by annealing temperature of 55°C. Simultaneously with detection of Kp16S rRNA gene, tetA and tetB genes were detected in 87.5% and 68.75% of tetracycline-resistant Klebsiella isolates, respectively. 56.25% of the isolates carried both tetA and tetB genes, simultaneously, and 12.5% of the isolates transferred one of the tetA or tetB genes.Conclusion: The results show that the multiplex PCR technique designed in this study is a fast, accurate, and sensitive method for identifying antibiotic-resistant K. pneumoniae that can be used for clinical samples or in epidemiological studies.
Keywords
Klebsiella pneumonia
Tetracycline
Multiplex polymerase chain reaction
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Journal of Isfahan Medical School
Volume 39, Issue 647 - Serial Number 647
September and October 2022
Pages 815-820

  • Receive Date 30 July 2021
  • Accept Date 06 April 2022