Detection of blaNDM-1 gene among the carbapenem resistant Escherichia coli and Klebsiella pneumoniae isolates from a children’s hospital in Nepal

Binod G.c ,Nim Raj Sapkota,Santosh Khanal,Pramod Poudel,Binod Rayamajhee,Sunil Lekhak,Jayram Lamichhane,Suraj Bahadur Thapa

doi:10.21608/nrmj.2018.17862

Abstract

This study was undertaken to detect the presence of blaNDM-1 gene among the carbapenem resistant Escherichiacoli and Klebsiella pneumoniae isolates, confirmed through using Modified Hodge Test (MHT). During the sixmonths of this study (November, 2016 – April, 2017), a total of 1503 clinical samples including urine, blood,wound swabs and Cerebrospinal fluid (CSF) were collected and processed at Microbiology Laboratory,International Friendship Children’s Hospital, Kathmandu, Nepal. Among the E. coli and K. pneumoniae isolates,14.1%, 17.1% were resistant to meropenem, whereas 11.28%, 17.1% were resistant to imipenem, respectively.From the pool of 94 E. coli and 35 K. pneumoniae isolates, 34 E. coli and 18 K. pneumoniae were screened aspossible carbapenemase producers. For screening of carbapenemase enzyme production among these isolates,resistances to third generation cephalosporins and carbapenems were taken into consideration. Two isolates of E.coli and 3 isolates of K. pneumoniae were confirmed as carbapenemase producers by MHT. Furthermore,Polymerase chain reaction (PCR) was carried out among the MHT positive bacteria for detection of blaNDM-1 gene.Genetic analysis of MHT positive isolates showed that 12 E. coli and 23 K. pneumoniae isolates were New Delhimetallo-β-lactamase producers (NDM-1). Results of this study demonstrated the presence of blaNDM-1 gene amongthe carbapenemase producing E. coli and K. pneumoniae isolates, and were recorded to be 50% and 66.6%,respectively.

Highlights

Carbapenems have been used for many years as the antibiotics of choice for treatment of nosocomial infections caused by Enterobacteriaceae
Since its first discovery at 2008 in a K. pneumoniae isolate recovered from a patient at a hospital, New Delhi, India, it has been transmitted to many species of Enterobacteriaceae in different countries (Yong et al, 2009)
Out of these samples processed, 270 (17.96%) showed bacterial growth. 151 bacterial cultures were isolated from urine samples, 65 from blood, 41 from pus, 13 from wound swabs, no bacterial isolates were recovered from Cerebrospinal fluid (CSF) (Table 1)

Summary

Introduction

Carbapenems have been used for many years as the antibiotics of choice for treatment of nosocomial infections caused by Enterobacteriaceae. Resistance to these drugs in Enterobacteriaceae has emerged worldwide mostly driven by the production of carbapenemases enzymes K. pneumoniae carbapenemase 2 (KPC-2) and more recently (NDM-1) (Padmini and Appalaraju, 2004). NDM-1 is one of the most important carbapenemases of Carbapenem Resistant Enterobacteriaceae (CRE). Yong et al, (2009); Deshpande et al, (2010) reported that NDM-1 attracted significant attention because the gene encoding this metallo beta lactamase (MBL) is located on a very mobile genetic element, the pattern of its spread proved to be more complex and apparently more unpredictable than the gene encoding K. pneumoniae carbapenemase (KPC)

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