Higher Prevalence of Extended Spectrum β-Lactamase Producing Uropathogenic Escherichia coli Among Patients with Diabetes from a Tertiary Care Hospital of Kathmandu, Nepal

Abstract

This study aimed to determine the occurrence of antibiotic resistance genes for β-lactamases; blaTEM and blaCTX-M in uropathogenic Escherichia coli isolates from urinary tract infection (UTI) suspected diabetic and nondiabetic patients. A hospital-based cross-sectional study was conducted in Kathmandu Model Hospital, Kathmandu, in association with the Department of Microbiology, GoldenGate International College, Kathmandu, Nepal, from June to December 2018. A total of 1,267 nonduplicate midstream urine specimens were obtained and processed immediately for isolation of uropathogens. The isolates were subjected to antibiotic susceptibility testing and extended spectrum β-lactamase (ESBL) confirmation. In addition, blaTEM and blaCTX-M genes were detected using specific primers. The overall prevalence of UTI was 17.2% (218/1,267), of which patients with diabetes were significantly more infected; 32.3% (31/96) as compared with nonpatients with diabetes, 15.9% (187/1,171). A total of 221 bacterial isolates were obtained from 218 culture-positive specimens in which E. coli was the most predominant; 67.9% (150/221). Forty-four percent (66/150) of the total E. coli was multidrug resistant and 37.3% (56/150) were ESBL producers. Among 56 isolates, 92.3% (12/13) were from patients with diabetes, and 83.0% (44/53) were from nondiabetics. Furthermore, 84.9% of the screened ESBL producers were confirmed to possess either single or both of blaTEM and blaCTX-M genes. The blaTEM and blaCTX-M genes were detected in 53.6% and 87.5% of the phenotypically ESBL confirmed E. coli, respectively. Higher rates of ESBL producing uropathogenic E. coli are associated among patients with diabetes causing an alarming situation for disease management. However, second-line drugs with broad antimicrobial properties are still found to be effective drugs for multidrug resistance strains.

One Health Assessment of an Urban Temporary Settlement Reveals Gut Microbiome Serving as Antimicrobial Resistance Gene Reservoir

“…Interestingly, phages and bacterial diversity were very similar in most of the samples, with human samples containing highest diversity (Table 1 and Supplementary table 1 and 2). This indicates that presence of any bacteria in an ecosystem, most likely harbors associated phages as well (Nayaju et al, 2021).…”

“…Some of these genes have been previously detected in Nepal while others like inuB, catQ, ksgA, floR , and bla EC (Table 3) were detected for the first time (Acharya et al, 2017; Joshi et al, 2017; Manandhar et al, 2020; Thakali et al, 2021a; Timsina et al, 2021). bla CTX_M and bla TEM were formerly detected from hospital samples (Manandhar et al, 2020)(Nayaju et al, 2021) as well as environmental samples (Thakali et al, 2020, 2021a). Similarly, qnrS, sul1 , and tetB (Thakali et al, 2021a; Young et al, 2022) were detected in samples of animal and environmental samples whereas ermB (Timsina et al, 2021) was detected from samples taken from school children in Nepal.…”

blaCTX-M Mediated Extended Spectrum Beta-lactamase Production in Escherichia coli from Urine Samples

“…PCR was done by initial denaturation at 95ºC for 15 minutes; followed by 35 cycles of denaturation at 94ºC for 45 seconds, annealing at 56ºC for 30 seconds, extension at 72ºC for 3 minutes and a fi nal extension at 72ºC for 10 minutes. The PCR products were then analyzed on 2% agarose gel electrophoresis (80V, 60 minutes) with 0.2 μg/mL concentration of ethidium bromide and then visualized by UV-transilluminator (Nayaju et al 2021).…”

“…After refrigeration, 16 μL of 1M of Tris-HCl was added and centrifugation was done at 8000 rpm for 2 minutes. The supernatant was collected and stored at -20°C (Gopal et al 2016).Conventional PCR technique was used to amplify CTX-M gene using primers F: 5' -TTTGCGATGTGCAGTACCAGTAA-3'; R: 5' -CTCCGCCTGCCGGTTTTAT-3'(Nayaju et al 2021).…”