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.
Navon-Venezia S, Kondratyeva K, Carattoli A. Klebsiella pneumoniae: A major worldwide source and shuttle for antibiotic resistance. FEMS Microbiol Rev 2017; 41(3): 252-75.
Effah CY, Sun T, Liu S, Wu Y. Klebsiella pneumoniae: An increasing threat to public health. Ann Clin Microbiol Antimicrob 2020; 19(1): 1.
Sathyavathy K, Madhusudhan BK. Review on clinical diseases caused by Klebsiella. J Pharm Res Int 2020; 32(21): 12-9.
Talebi Bezmin Abadi A, Rizvanov AA, Haertle T, Blatt NL. World Health Organization Report: Current crisis of antibiotic resistance. BioNanoScience 2019; 9(4): 778-88.
Canton R, Coque TM. The CTX-M beta-lactamase pandemic. Curr Opin Microbiol 2006; 9(5): 466-75.
Tuckman M, Petersen PJ, Howe AY, Orlowski M, Mullen S, Chan K, et al. Occurrence of tetracycline resistance genes among Escherichia coli isolates from the phase 3 clinical trials for tigecycline. Antimicrob Agents Chemother 2007; 51(9): 3205-11.
Bubpamala J, Khuntayaporn P, Thirapanmethee K, Montakantikul P, Santanirand P, Chomnawang MT. Phenotypic and genotypic characterizations of extended-spectrum beta-lactamase-producing Escherichia coli in Thailand. Infect Drug Resist 2018; 11: 2151-7.
MohammadAlipour Z, Asadpour L, Ranji N. Fluoroquinolone resistance and mutation in gyrA gene in clinical isolates of Klebsiella pnemoniae. Iran J Med Microbiol 2016; 10(5): 31-7. [In Persian].
Ding L, Yang Z, Lu J, Ma L, Liu Y, Wu X, et al. Characterization of phenotypic and genotypic traits of klebsiella pneumoniae from lung cancer patients with respiratory infection. Infect Drug Resist 2020; 13: 237-45.
Paknejad Z, Momtaz H, Tajbakhsh E. Determination of antibiotic resistance pattern in different serotypes of Klebsiella pneumoniae strains isolated from hospital infections in Zarinshahr. Applied Biology 2018; 8(29): 21-30. [In Persian].
Shu LB, Lu Q, Sun RH, Lin LQ, Sun QL, Hu J, et al. Prevalence and phenotypic characterization of carbapenem-resistant Klebsiella pneumoniae strains recovered from sputum and fecal samples of ICU patients in Zhejiang Province, China. Infect Drug Resist 2019; 12: 11-8.
Yu F, Lv J, Niu S, Du H, Tang YW, Pitout JDD, et al. Multiplex PCR analysis for rapid detection of Klebsiella pneumoniae carbapenem-resistant (Sequence Type 258 [ST258] and ST11) and hypervirulent (ST23, ST65, ST86, and ST375) strains. J Clin Microbiol 2018; 56(9).
Tajoadini M, Kheyrkhah B, Amini K. Detection of blaPER, blaGES and blaVEB genes in Shigella sonnei isolates from patients with diarrhea and determination of antibiotic susceptibility pattern. J Ardabil Univers Med Sci 2018; 18(1): 52-61. [In Persian].
Compain F, Babosan A, Brisse S, Genel N, Audo J, Ailloud F, et al. Multiplex PCR for detection of seven virulence factors and K1/K2 capsular serotypes of Klebsiella pneumoniae. J Clin Microbiol 2014; 52(12): 4377-80.
Naji-Hamad, M., & Nakhaei-Moghaddam, M. (2021). Detection of Tetracycline-Resistant Genes by Multiplex Polymerase Chain Reaction in Clinical Isolates of Klebsiella Pneumonia. Journal of Isfahan Medical School, 39(647), 815-820. doi: 10.22122/jims.v39i647.14366
MLA
Manar Naji-Hamad; Mahboobeh Nakhaei-Moghaddam. "Detection of Tetracycline-Resistant Genes by Multiplex Polymerase Chain Reaction in Clinical Isolates of Klebsiella Pneumonia", Journal of Isfahan Medical School, 39, 647, 2021, 815-820. doi: 10.22122/jims.v39i647.14366
HARVARD
Naji-Hamad, M., Nakhaei-Moghaddam, M. (2021). 'Detection of Tetracycline-Resistant Genes by Multiplex Polymerase Chain Reaction in Clinical Isolates of Klebsiella Pneumonia', Journal of Isfahan Medical School, 39(647), pp. 815-820. doi: 10.22122/jims.v39i647.14366
VANCOUVER
Naji-Hamad, M., Nakhaei-Moghaddam, M. Detection of Tetracycline-Resistant Genes by Multiplex Polymerase Chain Reaction in Clinical Isolates of Klebsiella Pneumonia. Journal of Isfahan Medical School, 2021; 39(647): 815-820. doi: 10.22122/jims.v39i647.14366
)