Extended Spectrum Beta-Lactamases (ESBLs) encoding genes (TEM, SHV and OXA) were amplified from multidrug resistance E. coli. The multidrug resistance E. coli isolates from different clinical sources were documented t...Extended Spectrum Beta-Lactamases (ESBLs) encoding genes (TEM, SHV and OXA) were amplified from multidrug resistance E. coli. The multidrug resistance E. coli isolates from different clinical sources were documented to be plasmid encoded and resistance against β-lactam and cephalosporin. Conventional laboratory analysis showed that seventy percent (70%) of the selected multidrug resistant clinical isolates were ESBLs positive, showing a ≥5 mm increase in zone diameter for either antibiotics compared to its zone when tested alone. The antibiotic susceptibility result showed that 100% of the isolates were resistant to amoxicillin-clavulanic acid, amoxicillin, cefuroxime and ampicillin-sulbactam while 90% of the isolates were resistant to ceftazidine and tetracycline, 80% to ofloxacin, 70% to ceftriazon, nalidixic acid, cefalexin, 60% to ciprofloxacin, 50% to nitrofurantoin, 40% to chloramphenicol and 20% to gentamicine. The multiplex PCR with primers TEM (931bp), SHV (868), OXA-2 (478), aac(3)-IIa (900) and rmtA (634), which are genes responsible for extended spectrum β-lactamase and aminoglycoside resistance in E. coli shows that: isolate W15 comprises of three (3) resistant gene, which corresponds with TEM resolving as a 931 base pair, SHV 868 base pair, and a 478 bp indicating OXA-2 that is faint probably indicating a low concentration of the gene. Isolate B2 comprises single resistant gene, which is interpreted as OXA-2 with 478 base pair while isolate URO2, U64 and S45 comprises of two resistance genes which resolve as 868 and 478 base pair indicating SHV and OXA-2 respectively. However, isolates S57, U58 and B7 showed no gene amplification despite the various degree of resistance in MIC and antibiotic susceptibility profile test obtained with conventional detection analysis. We assume that their resistant genes are not coded for by the primers used in this study as these isolates are likely to contain other resistant genes, which are also expressed at a molecular level. This study stands to show that molecular characterization has a great correlation with analytical methods.展开更多
This study was conducted to assess the occurrence and nature of extended-spectrum beta lactamase (ESBL) producing <em>Escherichia coli</em> and <em>Klebsiella pneumoniae</em> isolates from pati...This study was conducted to assess the occurrence and nature of extended-spectrum beta lactamase (ESBL) producing <em>Escherichia coli</em> and <em>Klebsiella pneumoniae</em> isolates from patients who presented with urinary tract infection at Federal Teaching Hospital Gombe. Isolates collected were recovered on MacConkey agar at 35<span style="white-space:nowrap;">°</span>C and were identified as members of Enterobacteriaceae, and further screened for antimicrobial susceptibility and resistance by disc diffusion method. Isolates resistant to oxyimino-cephalosporins were confirmed as ESBL producers using Double Disks Synergy Test (DDST). The study shows 66% resistance to ceftriaxone (30 μg) in <em>K. pneumoniae</em>, which was the highest value recorded and a 51% resistance to cefpodoxime (10 <em>μ</em>g) in <em>E. coli</em>. The sensitivity of <em>E. coli </em>and <em>K. pneumoniae</em> isolates to cefpodoxime (10 <em>μ</em>g) were 49% and 33.9% respectively. ESBLs were detected among 40% (40/100) of <em>E. coli</em> and 54.13% (59/109) of <em>K. pneumoniae</em> isolates. Molecular characterization of ESBL encoding genes among <em>E. coli</em> isolates using multiplex-PCR showed 10% prevalence of SHV gene and 5% prevalence for CTX-M gene while TEM gene was not detected. In <em>K. pneumoniae</em> isolates, 5% prevalence was recorded for each of the three genes screened. The study revealed a co-occurrence of SHV and CTX-M in 75% of the <em>E. coli</em> and 70% of the <em>K. pneumoniae</em> isolates;the occurrence of all the three genes was seen in 10% and 5% of <em>K. pneumoniae</em> and <em>E. coli</em> respectively. Multiplex-PCR method provided an efficient and rapid detection of ESBL related genes, hence could be used in epidemiological studies among ESBL isolates. Monitoring dissemination and transmissions of ESBL producers are highly recommended for optimum patient care and preventing the spread of multidrug resistant (MDR) pathogens.展开更多
文摘Extended Spectrum Beta-Lactamases (ESBLs) encoding genes (TEM, SHV and OXA) were amplified from multidrug resistance E. coli. The multidrug resistance E. coli isolates from different clinical sources were documented to be plasmid encoded and resistance against β-lactam and cephalosporin. Conventional laboratory analysis showed that seventy percent (70%) of the selected multidrug resistant clinical isolates were ESBLs positive, showing a ≥5 mm increase in zone diameter for either antibiotics compared to its zone when tested alone. The antibiotic susceptibility result showed that 100% of the isolates were resistant to amoxicillin-clavulanic acid, amoxicillin, cefuroxime and ampicillin-sulbactam while 90% of the isolates were resistant to ceftazidine and tetracycline, 80% to ofloxacin, 70% to ceftriazon, nalidixic acid, cefalexin, 60% to ciprofloxacin, 50% to nitrofurantoin, 40% to chloramphenicol and 20% to gentamicine. The multiplex PCR with primers TEM (931bp), SHV (868), OXA-2 (478), aac(3)-IIa (900) and rmtA (634), which are genes responsible for extended spectrum β-lactamase and aminoglycoside resistance in E. coli shows that: isolate W15 comprises of three (3) resistant gene, which corresponds with TEM resolving as a 931 base pair, SHV 868 base pair, and a 478 bp indicating OXA-2 that is faint probably indicating a low concentration of the gene. Isolate B2 comprises single resistant gene, which is interpreted as OXA-2 with 478 base pair while isolate URO2, U64 and S45 comprises of two resistance genes which resolve as 868 and 478 base pair indicating SHV and OXA-2 respectively. However, isolates S57, U58 and B7 showed no gene amplification despite the various degree of resistance in MIC and antibiotic susceptibility profile test obtained with conventional detection analysis. We assume that their resistant genes are not coded for by the primers used in this study as these isolates are likely to contain other resistant genes, which are also expressed at a molecular level. This study stands to show that molecular characterization has a great correlation with analytical methods.
文摘This study was conducted to assess the occurrence and nature of extended-spectrum beta lactamase (ESBL) producing <em>Escherichia coli</em> and <em>Klebsiella pneumoniae</em> isolates from patients who presented with urinary tract infection at Federal Teaching Hospital Gombe. Isolates collected were recovered on MacConkey agar at 35<span style="white-space:nowrap;">°</span>C and were identified as members of Enterobacteriaceae, and further screened for antimicrobial susceptibility and resistance by disc diffusion method. Isolates resistant to oxyimino-cephalosporins were confirmed as ESBL producers using Double Disks Synergy Test (DDST). The study shows 66% resistance to ceftriaxone (30 μg) in <em>K. pneumoniae</em>, which was the highest value recorded and a 51% resistance to cefpodoxime (10 <em>μ</em>g) in <em>E. coli</em>. The sensitivity of <em>E. coli </em>and <em>K. pneumoniae</em> isolates to cefpodoxime (10 <em>μ</em>g) were 49% and 33.9% respectively. ESBLs were detected among 40% (40/100) of <em>E. coli</em> and 54.13% (59/109) of <em>K. pneumoniae</em> isolates. Molecular characterization of ESBL encoding genes among <em>E. coli</em> isolates using multiplex-PCR showed 10% prevalence of SHV gene and 5% prevalence for CTX-M gene while TEM gene was not detected. In <em>K. pneumoniae</em> isolates, 5% prevalence was recorded for each of the three genes screened. The study revealed a co-occurrence of SHV and CTX-M in 75% of the <em>E. coli</em> and 70% of the <em>K. pneumoniae</em> isolates;the occurrence of all the three genes was seen in 10% and 5% of <em>K. pneumoniae</em> and <em>E. coli</em> respectively. Multiplex-PCR method provided an efficient and rapid detection of ESBL related genes, hence could be used in epidemiological studies among ESBL isolates. Monitoring dissemination and transmissions of ESBL producers are highly recommended for optimum patient care and preventing the spread of multidrug resistant (MDR) pathogens.
