Molecular detection of β-lactamase and integron genes in clinical strains of Klebsiella pneumoniae by multiplex polymerase chain reaction

Abstract

Introduction:Infections caused by β-lactamase-producing gram-negative bacteria, such as Klebsiella pneumoniae, are increasing globally with high morbidity and mortality. The aim of the current study was to determine antimicrobial susceptibility patterns and the prevalence of antibiotic resistance genes (β-lactamase and integron genes) using multiplex PCR. Methods: One-hundred K. pneumoniae isolates were collected from different clinical samples. Antibiotic susceptibility testing was performed with thirteen different antibiotics. Multiplex-PCR was used to detect β-lactamase (bla TEM , bla CTX-M, bla SHV, bla VEB , bla PER , bla GES , bla VIM, bla IMP, bla OXA , and bla KPC ) and integron genes (int I, int II, and int III). Results: The highest and lowest rate of resistance was exhibited against amikacin (93%) and imipenem (8%), respectively. The frequency of β-lactamase-positive K. pneumoniae was 37%, and the prevalence of the bla TEM , bla CTX-M, bla SHV, bla VEB , bla PER , bla GES , bla VIM, bla IMP, bla OXA , and bla KPC genes was 38%, 24%, 19%, 12%, 6%, 11%, 33%, 0%, 28%, and 23%, respectively. Of the 100 isolates, eight (8%) were positive for class I integrons; however, class II and III integrons were not detected in any of the strains. Conclusions: These results indicate co-carriage of a number of β-lactamase genes and antibiotic resistance integrons on the same plasmids harboring multi-drug resistance genes. It seems that these properties help to decrease treatment complications due to resistant bacterial infections by rapid detection, infection-control programs and prevention of transmission of drug resistance.

Prevalence of extended spectrum beta lactamase and plasmid mediated quinolone resistant genes in strains of Klebsiella pneumonia, Morganella morganii, Leclercia adecarboxylata and Citrobacter freundii isolated from poultry in South Western Nigeria

“…TEM gene was the most prevalent ESBL gene detected followed by SHV and CTX-M gene in clinical isolates ( Machado et al, 2007 ; Sedighi et al, 2017 ). Fortini et al (2011) has also reported dominance of bla (TEM) gene in Enterobacteriaceae isolated from Nigeria and only one strain was positive for the CTX-M gene.…”

Characterization of β-lactam resistance in K. pneumoniae associated with ready-to-eat processed meat in Egypt

“…A study conducted by Sedighi et al . showed that Int I was present in 8% of isolates while Int II and Int III were absent from all isolates [ 47 ]. Rezai et al .…”

“…Both Int I and Int III were co-founded in 20% of isolates and 10% of isolate co-carried Int II and Int III. A study conducted by Sedighi et al showed that Int I was present in 8% of isolates while Int II and Int III were absent from all isolates [47]. Rezai et al reported that Int I was detected in 79.3% while Int II was harbored by 10.3% of isolates [48].…”

Occurrence of Extended Spectrum Beta Lactamase Encoding Genes among Urinary Pathogenic <i>Escherichia coli</i> and <i>Klebsiella pneumoniae</i> Isolates Obtained from a Tertiary Hospital in Gombe Nigeria

“…Multiplex-PCR was performed using PTC-100 thermal cycler (USA) to detect beta lactamase genes (blaTEM, blaCTX-M and blaSHV) using PCR conditions as described by [39]. The M-PCR was carried out using 1.5 µl of extracted genomic DNA in a 20 µl PCR reaction mixture consisting of 2.5 µl 10× PCR buffer, 1.5 µl MgCl 2 (50 mM), 0.5 µl dNTPs (10 mM), 1.5 µl of each primer, 0.5 µl of Taq DNA polymerase, and 9 µl sterile distilled water.…”