First Sequence Analysis of Genes Mediating Extended-Spectrum Beta-Lactamase (ESBL) bla-TEM, SHV- and CTX-M Production in Isolates of Enterobacteriaceae in Southern Benin

Abstract

The production of extended-spectrum beta-lactamase (ESBL) by Enterobacteriaceae is a global public health problem. The present study was carried out on 156 strains of enteric bacteria isolated from urinary and cervicovaginal fluid samples. Identification of the strains was performed using MALDI-TOF MS and antibiotic susceptibility tests by disk diffusion method on Mueller Hinton agar in accordance with the recommendations of the Antibiogram Committee of the French Society for Microbiology. ESBL genes were sought by real time-polymerase chain reaction (RT-PCR) and by gel-based PCR. Gel-based PCR products were used for sequencing of the resistance genes, which were analyzed in the NCBI and Arg Annot databases. Results showed a predominance of Escherichia coli both in the urinary and cervicovaginal fluid samples. Klebsiella pneumoniae was the second most isolated bacterium in the specimens. Sensitivity to antibiotics revealed high levels of cephalosporin resistance but low resistance to carbapenem. No resistance was noted to colistine. The bla-TEM gene was present in Escherichia coli, while bla-SHV was found in Klebsiella pneumoniae and bla-CTX-M was recovered in both strains. Analysis of the sequences revealed that bla-Tem1 was predominant in bla-TEM and bla-CTX-M-15 was most represented by bla-CTX-M. This study confirms the presence of ESBL-producing Enterobacteria in Benin. This was an epidemiological study aimed at detecting cephalosporin resistance in gram-negative Bacillus isolated from urinary tract and genital infections developed by women. Since the advent of molecular biology techniques for the identification of resistance in bacteria including determination of ESBL resistance genes (i.e., TEM, SHV, CTX-M), no study has been conducted to identify the different variants that circulate in Benin by sequencing these resistance genes. This sequencing is essential in order to differentiate the non-ESBL parental enzymes, which is not possible with the commonly used PCR techniques that do not permit differentiation of the point generating different variants of the ESBL genes. The present study then helped to identify those variants, in particular Tem1, SHV1, and CTX-M15, which are most encountered in Benin and around the world.

Prevalence of Escherichia coli and Klebsiella pneumoniae, Producing Extended-Spectrum Beta-Lactamase (ESBLs) from Clinical Specimen in Khuzestan, Iran

“…pneumoniae and E. coli are the members of the Enterobacteriaceae family that cause nosocomial infections. Recently, in many parts of the world, antibiotic resistance due to the presence of beta-lactamase enzymes increased against a wide variety of antibiotics (20)(21)(22). In this study, the results indicate a high rate of antibiotic resistance, which is consistent with the results of many studies around the world, but in some studies, this rate of resistance is low, which indicates differences in the pattern of antibiotic use in different regions (23,24).…”

Characterization of Potential Pathogenic Bacteria Isolated in High-Risk Infectious Services at the University Hospital Center of Suru-Léré in Benin

“…The same finding was made by Maina et al, who found in Nairobi, Kenya, that the presence of the SHV gene could predict resistance to Ceftazidime [26]. The coexistence of different resistance genes in gram-negative bacteria may explain their MDR [27] [28]. Therefore, the hospital environment (air, soil, surfaces, equipment) contaminated by MDR bacteria can serve as a reservoir for crosstransmission of these bacteria [29].…”

Prescription of antibiotics as a source of emerging antibiotic resistance: Knowledge, attitudes, and practices of medical staff in the Dassa-Glazoué and Savalou-Bantè's health zones (Benin, West Africa)

“…In the Savalou-Bantè health zone, there is a great use of carbapenem which is normally used in case of resistance to the third-generation cephalosporins. This observation explains the emergence of resistance through the production of BLSE and carbapenem observed in Benin [13][14][15][16]. It is also well known that the use of third-generation cephalosporins is responsible for the slight reduction in carbapenem resistance [17].…”