Abstract
Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the most prominent bacterial species resistant to almost all commonly used antibiotics. Carbapenem is one of the last resort drugs for treating such emerging multidrug-resistant bacteria. This study aimed to detect carbapenem-resistant blaNDM-1 gene in ESBL producing E. coli and K. pneumoniae isolates. A total of 190 E. coli and 350 K. pneumoniae isolates were screened for extended spectrumβ-lactamase (ESBL), carbapenemase and metallo β-lactamase (MBL) production via double-disk synergy test (DDST), modified Hodge test and combined-disk diffusion method. The blaNDM-1 gene was detected by PCR and confirmed via Sanger sequencing method. Of the 540 isolates tested, 71.8% were found to be multidrug-resistant. Overall rate of ESBL-positive isolates were 57.89% E. coli and 31.42% K. pneumoniae. Among ESBL positive isolates, 49.09% E. coli and 40% K. pneumoniae were positive for carbapenemase production whereas MBL production was detected in 29% E. coli and 22% K. pneumoniae isolates. In MBL positive isolates, (37%) E. coli and (40%) K. pneumoniae isolates harboured blaNDM-1 gene. The pair-wise DNA was aligned with the NDM-1 sequence from GenBank. The alignment score was 243 and the blast nucleotide sequencing results showed 97% sequence similarity with the sequences in GenBank for the blaNDM-1 gene. The blaNDM-1 gene was found to be the most prevalent in urine samples. There is a dire need to conduct screening tests in hospitals and communities to find out the exact prevalence of the blaNDM-1 spread in our population.
Highlights
Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the most prominent bacterial species resistant to almost all commonly used antibiotics
Out of 540 clinical isolates collected from different hospitals in Peshawar and Islamabad, 190 (35.2%) were identified as E. coli and 350 (64.8%) as K. pneumoniae being the most predominant species
Antimicrobial susceptibility In E. coli isolates, highest resistance was observed towards ceftriaxone (74.7%), followed by ampicillin (74.2%) and cefotaxime (69.4%), whereas imipenem (94.7%), doripenem (88.4%), and nitrofurantoin (86.8%) are found to be the most effective drugs in the present study
Summary
Enterobacteriaceae such as Escherichia coli and Klebsiella pneumoniae are the most prominent bacterial species resistant to almost all commonly used antibiotics. This study aimed to detect carbapenem-resistant blaNDM-1 gene in ESBL producing E. coli and K. pneumoniae isolates. In MBL positive isolates, (37%) E. coli and (40%) K. pneumoniae isolates harboured blaNDM-1 gene. New Delhi Metallo-β-lactamase (NDM-1) type metallo-beta lactamase (MBL) is one of the most potent and prevalent enzymes of MBL in Escherichia coli, Klebsiella pneumoniae and other Gram-negative rods [1]. The gene was identified in a 180kb plasmid [3,4], but later on, it was discovered that other plasmids ranged from 50-500 kb of various Gram-negative species, harbored this gene [5]. The blaNDM-1 gene is present on various plasmids, containing other resistant genes such as ESBL genes (CTX-M-15, SHV-12), carbapenem-resistant genes or carbapenemase-encoding genes (OXA-48, OXA-181, VIM), plasmid-associated cephalosporinase genes (CMY-16, CMY-58), aminoglycoside resistance genes (16s rRNA methylase), macrolide resistance genes (esterase), rifampin resistance gene, qnr genes (qnrAB, qnrB1, qnrB2) and sulfamethoxazole resistance genes, reported from other parts of the world including Pakistan [6,7,8,9]
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