Home About Journal AHEAD OF PRINT Current Issue Back Issues Instructions Submission Search Subscribe Blog    

Users Online: 1364 
Print this page  Email this page Small font sizeDefault font sizeIncrease font size 

 Table of Contents    
Year : 2014  |  Volume : 48  |  Issue : 5  |  Page : 533-535
Klebsiella pneumoniae carrying blaNDM-1 gene in orthopedic practice

1 Department of Microbiology, Government Medical College Hospital, Sector 32, Chandigarh, India
2 Department of Orthopedics, Government Medical College Hospital, Sector 32, Chandigarh, India

Click here for correspondence address and email

Date of Web Publication2-Sep-2014


Emergence and spread of carbapenemases in Enterobacteriaceae is a cause of concern worldwide, the latest threat being New Delhi metallo-β-lactamase (NDM-1). This report is of an orthopedic case with fracture femur managed with internal fixation and bone grafting, who subsequently developed secondary infection with Klebsiella pneumoniae harboring blaNDM-1 gene. Minimum inhibitory concentration (MIC) of imipenem was ≥8 μg/ml by E-test, suggestive of carbapenemase production. Phenotypic and further genotypic detection confirmed the presence of blaNDM-1 gene. The isolate remained susceptible only to tigecycline, colistin, and polymyxin B.

Keywords: Carbapenemase, Klebsiella pneumoniae, New Delhi metallo-β-lactamase
MeSH terms: Klebsiella, orthopedic surgery, betalactamase

How to cite this article:
Gupta V, Bansal N, Gupta R, Chander J. Klebsiella pneumoniae carrying blaNDM-1 gene in orthopedic practice. Indian J Orthop 2014;48:533-5

How to cite this URL:
Gupta V, Bansal N, Gupta R, Chander J. Klebsiella pneumoniae carrying blaNDM-1 gene in orthopedic practice. Indian J Orthop [serial online] 2014 [cited 2020 Feb 26];48:533-5. Available from:

   Introduction Top

Resistance to antimicrobials and a paucity of new antimicrobial agents are ongoing challenges, more so in orthopaedic surgery. Carbapenems are one of the very few therapeutic agents available for treating multidrug resistant infections caused by Enterobacteriaceae; however, carbapenemases are increasingly being reported, with the latest threat being New Delhi metallo-β-lactamase (NDM-1). NDM-1 was initially reported in Klebsiella pneumoniae and  Escherichia More Details coli in a Swedish patient of Indian origin, [1] and several published articles, particularly case reports, show the presence of NDM-1 containing pan-resistant Enterobacteriaceae mainly from the Indian subcontinent. [2],[3],[4],[5],[6] However, now NDM-1 carriage has spread in enterobacterial isolates the world over, which is a matter of great concern. [7] This is the report of an orthopedic case with fracture femur managed with internal fixation and bone grafting, who subsequently developed secondary infection with K. pneumoniae harboring blaNDM-1 gene.

   Case Report Top

A 32-year-old male patient, presented with a Grade IIIA open fracture of the right distal femur, following a roadside accident suffered 22 hours back. Local examination revealed two (sutured) lacerated wounds on the distal one-third of the anterior and lateral aspects of the right thigh. There were no other injuries, and the distal neurovascular function was intact.

After stabilization of the patient, he was taken up for surgery after 24 hours of admission (46 hours of injury). Through a lateral surgical approach (incorporating the existing wounds), the wounds were debrided, intercondylar fracture was stabilized with K wires and lag screws, and the distal femur fracture was stabilized with a locking plate. At the time of surgery, there was significant bone loss from the lateral and posterior parts of the distal femur, but bone grafting was not done due to the fear of infection in the contaminated wound which was being operated after a delay of 46 h. Wound was closed on a negative suction drain. The patient was put on gentamicin and cefoperazone/sulbactum. One week later, the wound was apparently healthy, so the patient underwent bone grafting at the fracture site. Intraoperatively, fluid and tissue from the fracture site were sent for culture, which came out to be sterile.

On the third day of the second surgery, the wound showed seropurulent discharge which on culture/sensitivity yielded pure growth of multidrug resistant K. pneumoniae. Since the organism was highly resistant, the patient was put on amikacin (1 g once daily), high-dose imipenem (1 g 8-hourly as infusion in normal saline over 2 h), and rifampicin (600 mg once daily). A repeat pus culture after 2 days again revealed the same organism. A decision for wound debridement and removal of bone graft was made and performed which resulted in healing of the wound. No further colonization with the same bacterium was detected by taking rectal swabs from that patient or from other patients hospitalized simultaneously in the same unit.

In the laboratory, K. pneumoniae isolate was found to be multidrug resistant by disk diffusion method as per the Clinical Laboratory Standards Institute (CLSI) 2011 guidelines. [8] It was resistant to ceftazidime, cefotaxime, cefipime, gentamicin, amikacin, ciprofloxacin, aztreonam, amoxicillin/clavulanic acid, cefoperazone/sulbactam, piperacillin/tazobactam, ampicillin/sulbactam, cefipime/tazobactam, ertapenem, meropenem, doripenem, and imipenem. We screened the isolate for extended-spectrum β-lactamase (ESBL) production by disk potentiation test, but it was found to be negative. Evaluation of the isolate for ESBL using E-test strips containing ceftazidime at one end and ceftazidime-clavulanic acid at the other end, as well as Amp C β-lactamase by using E-test strips containing cefotetan at one end and cefotetan/cloxacillin at the other end was also negative. Minimum inhibitory concentration (MIC) of imipenem was ≥8 μg/ml by E-test, suggestive of carbapenemase production. Modified Hodge test on the isolate was negative, ruling out class A carbapenemases and pointing toward metallo-β-lactamases (MBLs). Phenotypic method with disk synergy test using imipenem and ethylenediaminetetraacetic acid (EDTA) and further genotypic detection confirmed the presence of blaNDM-1 gene [Figure 1] in hospital-acquired K. pneumoniae isolate, conferring resistance to carbapenems including doripenem. The isolate remained susceptible only to tigecycline (MIC ≤1.5 μg/ml), colistin (MIC ≤1 μg/ml), and polymyxin B (MIC ≤1 μg/ml).
Figure 1: Pulsed-field gel electrophoresis pattern of NDM-1 producing Klebsiella pneumoniae

Click here to view

Molecular analysis technique

DNA was extracted from the strain by heat boil method, and the DNA was subjected to single-target polymerase chain reaction (PCR). Two microliters of the extracted total DNA was subjected to PCR in a 50-μl reaction mixture. The PCR mixture for the detection of MBL genes contained 1 × PCR buffer [10 mM Tris-HCl (pH 8.3), 50 mM KCl], 1.5 mM MgCl2, 0.125 mM of each deoxynucleotide triphosphate, 0.1 μM of each primer, and 2 U of DNA polymerase. The primers used were F: 5′-GGGCAGTCGCTTCCAACGGT-3′ and R: 5′-GTAGTGCTCAGTGTCGGCAT-3′, which amplified an internal fragment of 475 bp for NDM-1 gene. Amplification was carried out under the following thermal cycling conditions: 10 min at 94°C; 36 cycles of amplification consisting of 30 s at 94°C, 40 s at 52°C, and 50 s at 72°C; and 5 min at 72°C for the final extension. Amplified products (475 bp) from the test strain and control strain were visualized under UV light on 3% agarose gel electrophoresis [Figure 1]. [9]

   Discussion Top

Acquired carbapenemases confer extensive antibiotic resistance in Enterobacteriaceae and represent a public health threat. A novel acquired carbapenemase, NDM-1, has now been described from several nations worldwide, [7] and was earlier being reported mostly in patients who had received treatment in the Indian subcontinent. [2],[3],[4],[5],[6] Like other acquired MBLs, NDM-1 hydrolyzes all β-lactam antibiotics except aztreonam.

NDM-1, named after the city of origin (New Delhi) and which has been recently criticized, is normally carried on a variety of plasmids along with genes conferring resistance to almost all other antibiotics. This enzyme shares very little identity with other MBLs, with the most similar MBLs being VIM-1/VIM-2, with which it has only 32.4% identity. [1] Most plasmids detected in these clinically relevant bacteria are easily transferable and capable of wide rearrangement, suggesting rapid and global dissemination and malleability among bacterial populations. [3] An association with other resistance mechanisms makes a majority of Enterobacteriaceae with blaNDM-1 extensively resistant to antibiotics and susceptible only to polymyxins and less consistently to tigecycline; similar was the finding in our isolate.

K. pneumoniae is a well known hospital-acquired pathogen, notorious for being multidrug resistant. Castanheira et al. [10] reported that isolates producing NDM-1 were disseminated in Indian health care facilities as early as 2006, but reports of a clinical association of K. pneumoniae carrying blaNDM-1 gene responsible for wound infections are rare. Recently there has been a report on diabetic foot ulcer patients. [11] To treat these infections, combining drugs from different classes of antibiotics is to be resorted to or the pus/foreign agent should be removed as was done in the present case.

Invasive infections by carbapenem-resistant strains are associated with high morbidity and mortality rates and such case reports alert the surgeons and physicians about the threat of such multidrug-resistant strains. To conclude, infections by carbapenem-resistant bacteria are difficult to treat and rapid identification of MBL-producing Gram-negative species is crucial both for appropriate treatment and for timely implementation of infection control measures.

   Acknowledgments Top

We are thankful to Dr. Pallab Ray, Dr. Vikas Gautam, and their team from the Department of Medical Microbiology, PGIMER, Chandigarh for helping us with the molecular work and providing us the control strain for molecular analysis.

   References Top

1.Yong D, Toleman MA, Giske CG, Cho HS, Sundman K, Lee K, et al. Characterization of a new metallo-beta-lactamase gene, blaNDM-1 and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother 2009;53:5046-54.  Back to cited text no. 1
2.Wu HS, Chen TL, Chen IC, Huang MS, Wang FD, Fung CP, et al. First Identification of a patient colonized with Klebsiella pneumonia carrying blaNDM-1 in Taiwan. J Chin Med Assoc 2010;73:596-8.  Back to cited text no. 2
3.Chihara S, Okuzumi K, Yamamoto Y, Oikawa S, Hishinuma A. First case of New Delhi metallo- b- lactamase 1 producing Escherichia coli infection in Japan. Clin Infect Dis 2011;52:152-5.  Back to cited text no. 3
4.Mulvey MR, Grant JM, Plewes K, Roscoe D, Boyd DA. New Delhi metallo-beta-lactamase in Klebsiella pneumonia and Escherichia coli, Canada. Emerg Infect Dis 2011;17:103-6.  Back to cited text no. 4
5.Zarfel G, Hoenigl M, Leitner E, Salzer HJ, Feierl G, Masoud L, et al. Emergence of New Delhi metallo- beta- lactamase, Austria. Emerg Infect Dis 2011;17:129-30.  Back to cited text no. 5
6.Sidjabat H, Nimmo GR, Walsh TR, Binotto E, Htin A, Hayashi Y, et al. Carbapenem resistance in Klebsiella pneumonia due to New Delhi metallo- beta-lactamase. Clin Infect Dis 2011;52:481-4.  Back to cited text no. 6
7.Notes from the field: Hospital outbreak of carbapenem-resistant Klebsiella pneumoniae producing New Delhi metallo-beta-lactamase--Denver, Colorado, 2012. Centers for Disease Control and Prevention (CDC). MMWR Morb Mortal Wkly Rep 2013;62:10.  Back to cited text no. 7
8.Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. Twentieth informational supplement. Document M100-S20. Wayne, PA: CLSI; 2010.  Back to cited text no. 8
9.Deshpande P, Rodrigues C, Shetty A, Kapadia F, Hedge A, SomanR. New Delhi Metallo-beta lactamase (NDM-1) in Enterobacteriaceae: Treatment options with carbapenems compromised. J Assoc Physicians India 2010;58:147-9.  Back to cited text no. 9
10.Castanheira M, Deshpande LM, Mathai D, Bell JM, Jones RN, Mendes RE. Early dissemination of NDM-1- and OXA-181- producing Enterobacteriaceae in Indian hospitals: Report from the SENTRY Antimicrobial Surveillance Program, 2006-2007. Antimicrob Agents Chemother 2011;55:1274-8.  Back to cited text no. 10
11.Khan AU, Nordmann P. NDM-1 producing Enterobacter cloacae and Klebsiella pneumoniae from diabetic foot ulcer patients in India. J Med Microbiol 2012;61:454-6.  Back to cited text no. 11

Correspondence Address:
Varsha Gupta
Department of Microbiology, Government Medical College Hospital, Sector 32 B, Chandigarh - 160 030
Login to access the Email id

Source of Support: Department of Microbiology, Government Medical College Hospital, Chandigarh, Conflict of Interest: None

DOI: 10.4103/0019-5413.139888

Rights and Permissions


  [Figure 1]

This article has been cited by
1 Anti-infective therapy without antimicrobials: Apparent successful treatment of multidrug resistant osteomyelitis with hyperbaric oxygen therapy
Elsa Goerger,Estelle Honnorat,Hélène Savini,Mathieu Coulange,Eric Bergmann,Fabrice Simon,Piseth Seng,Andreas Stein
IDCases. 2016; 6: 60
[Pubmed] | [DOI]


    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Email Alert *
    Add to My List *
* Registration required (free)  

   Case Report
    Article Figures

 Article Access Statistics
    PDF Downloaded90    
    Comments [Add]    
    Cited by others 1    

Recommend this journal