Phenotypic and genotypic detection of extended spectrum β-lactamases among Escherichia coli and Klebsiella pneumoniae isolates from type 2 diabetic patients with urinary tract infections

Abstract

Background: T2DM patients are more likely to have UTIs caused by resistant organisms such as ESBLs producing bacteria. Challenging reliable identification and prompt characterization of in-vitro susceptibilities of these bacteria are the first steps of deciding the appropriate antimicrobial therapy for UTIs caused by them.
 Objectives: To isolate and identify E. coli and K. pneumoniae from urine of T2DM patients with UTIs, to determine antibiotic resistance pattern among isolates, and to identify ESBLs production phenotypically and genotypically.
 Material and method: All samples were cultured on Cystine-Lactose-Electrolyte-Deficient Agar medium (CLED) by using calibrated loop. Growth of 100 colonies or more, i.e. 105 colony forming units (CFU)/mL urine was considered as signifi- cant bacteriuria. Isolation and identification were done according to standard method. All isolates were tested for antibiotic susceptibility testing by the disc diffusion method according to CLSI guidelines. Phenotypic detection of ESBLs was done by double-disk synergy test. Genotypic detection of blaTEM, blaSHV and blaCTX-M genes by using PCR.
 Results: Results of this study showed that E. coli and K. pneumoniae were the dominant bacterial isolates, they constituted 103 (91.2%) out of 113 urine isolates. E. coli (58. 4%) K. pneumoniae (32.7%), Enterococcus spp. (4.4%), Proteus spp. (2.7%) and Pseu- domonas spp. (1.8%). About 25 (24.3%) out of 103 E. coli and K. pneumoniae isolates were ESBLs positive by DDST, and 22 (88.0%) out of them had ESBLs encoding genes by conventional PCR. The most common gene detected was blaTEM (59.1%), followed by blaSHV (27.3%). CTX-M had not been detected in any of testes isolates.
 Conclusion: blaTEM and blaSHV genes were detected in 22 out of 25 ESBLs producing E. coli and K. pneumoniae isolates phenotypically detected by DDST. blaTEM was found to be the predominant gene (59.1%), while blaCTX-Mene was not detected in any of tested isolates.
 Keywords: Extended Spectrum β-Lactamases; Type 2 diabetes mellitus; Urinary tract infections; Phenotypic; genotypic methods.

Prevalence of β-lactamase enzyme producing uropathogenic E.coli isolates amongst diabetic and non-diabetic patients

“…Prevalence of ESBL coding genes among diabetic isolates in the present investigation were higher than one of the similar study reports that 58.2% of phenotypically ESBL producers among these 26.6% harbor blaTEM and 12.5% harbor blaCTX-M among diabetic UPEC [38]. ESBL production in Type 2 diabetic UPEC (25.8%) were positively correlated with old age, urinary catheterization, discriminate use of antibiotics and acute kidney injury [24].…”

“…Current reports of ESBL production were correlated with the reports of [24] that 25.8% of ESBL prevalence in Type 2 diabetic E.coli. Findings of [17] revealed ESBL-producing E.coli isolates were significantly more resistant to antibiotics as compared to non-producers of ESBL.…”

Synergy of Plant Essential Oils in Antibiotic Therapy to Combat Klebsiella pneumoniae Infections

“…Oils were used at subinhibitory concentration dilutions (one quarter of the MIC was used), while antibiotics were used at the gradients described in the Materials and Methods section. Then, the assay was performed as previously described [ 51 ]. The fractional inhibitory concentration (FIC) for each EO and antibiotic was calculated by dividing the MIC of two drugs in combination with the MIC of each drug alone.…”

Editorial: Infectious diseases and NCDs persist despite concerted effort

“…El- Domany and colleagues 1 report carbapenem resistant genes in Klebsiella pneumonia. It is followed by an interesting paper on genotypic detection of extended spectrum β-lactamases among Escherichia coli and Klebsiella pneumoniae isolates from type 2 diabetic patients with urinary tract infections keeping the Klebsiella theme alive 2 . Number 3 goes to: yes, Klebsiella again!…”