Prevalence of ESBL and MBL antibiotic resistance genes in Klebsiella
pneumoniae in Pretoria Academic Hospital
Maningi NE, Ehlers MM, Hoosen AA, Makgothlo E, Omar S and Kock MM
Department of Medical Microbiology, Faculty of Health Sciences, University of Pretoria/NHLS
INTRODUCTION
• Extended spectrum-β-lactamase (ESBL) are group of βlactamases that are plasmid mediated that have the ability to
RESULTS AND DISCUSSION
• Multiplex PCR detected the presence of CTX-M, SHV and TEM
genes in the K. pneumoniae isolates from the Pretoria
hydrolyse the newer cephalosporins and aztreonam and are
inhibited by clavulanic acid
• Extended spectrum-β-lactamase producing K. pneumoniae is
the cause of most of the multi-drug resistant nosocomial
Academic Hospital (Figure 1)
• No VIM genes were detected with the multiplex PCR in the K.
pneumoniae isolates included in this study
L
1
2
3
4
5
6
7
8
9
10 11
12 13
14
15
C
L
infections
• The ESBL-producing K. pneumoniae can be identified by the
747 bp
blaSHV
593 bp
blaCTX-M
presence of the genes blaCTX-M, blaSHV and blaTEM coding
100 bp DNA
ladder
for the β-lactamases cef enzymes
• The transmission of ESBLs is due to clonal spread, plasmid
dissemination or both mechanisms among or between
445 pb
blaTEM
species
• Metallo-β-lactamases has recently emerged as a resistant
Figure 1:
class of broad-spectrum β-lactamases in K. pneumoniae and
it can be identified by the presence of blaVIM gene
• These ESBLs and MBL enzymes can be detected
AIM PCR assay
simultaneously using a multiplex
• This study investigated the prevalence of ESBL and MBL
antibiotic resistance genes in K. pneumoniae isolates from
Multiplex PCR simultaneously amplified the blaCTXM (593 bp),
blaSHV (747 bp)
and blaTEM (445 bp). In lanes 2, 5, 6 7, 8, 9 10, 11, 12,13 and 14 ESBL genes were
detected while lanes 1, 3, 4, and 15 were negative for all three genes. Negative control
was included in lane C and lanes L represent the molecular weight marker (100 bp
DNA ladder, Promega, Medison, USA)
•The ESBL genes were detected in different combinations as only
one gene, two genes or all three genes in one isolate (Figure 2)
• Most of the ESBL-producing K. pneumoniae isolates
harboured three ESBL genes (Figure 2)
Pretoria Academic Hospital
MATERIALS AND METHODS
SHV, CTX, TEM, 36%
SHV, TEM, 3%
CTX, TEM, 18%
SHV, 16%
TEM, 4%
N, 13%
VIM 0
CTX, SHV 3%
CTX-M, 3%
• This study was a prospective descriptive study in which
ninety seven K. pneumoniae isolates were obtained from
clinical specimens sent for analysis to the Microbiology
laboratory, National Health Laboratory Services, Pretoria
Academic Hospital
• The VITEK 2 (bioMérieux, France) was used for phenotypic
• identification of the isolates as ESBL-producing K.
Figure 2:
Pie chart representation of Multiplex PCR results of three ESBL genes blaCTX-M,
blaSHV and blaTEM detected in bacterial isolates
• Multiplex PCR results obtained in this study of K. pneumoniae
pneumoniae
• The MagnaPure LC Compact (Roche Applied Science,
Germany) was used for the automated extraction of DNA
according to manufacturer’s instructions
• The multiplex PCR assay amplifying the blaCTX-M, blaSHV,
blaTEM and blaVIM genes was performed using the QIAGEN
Multiplex PCR kit (QIAGEN, USA) following the manufacture’s
isolates revealed that 89% (87/97) were positive for ESBL genes
• This prevalence of ESBL-producing K. pneumoniae genes
is alarming compared to the 22% prevalence of these gene in
SA 1998-1999 (Michel et al., 2006)
• Carbapenems are the preferred antibiotic for the treatment of
serious infections caused by ESBLs, since they are resistant to
the newer expanded-spectrum cephalosporins
instructions
REFERENCES
• Aranzazu Valverde, Teresa MC, Lucia GM, Fernando B and Rafael C (2008) Complex molecular epidemiology of
extended-spectrum β-lactamases in Klebsiella pneumoniae: a long-term perspective from a single intitution in
Madrid. Journal of antimicrobial Chemotherepy 61:64-72
• Franceco L, Jean-Denis D, Celine C, Adrea E, Gianluigi L, Gianfranco A, Gian M and Antoniolo (2004) Emergence
in K. pneumoniae and E. cloacae clinical isolates of the VIM-4 Mettallo-β-Lactamase encoded by a conjugative
plasmid. Antimicrobial agents and chemothetapy 48:648-650
• Michel A, Pfaller and John S (2006) Overview of the epidemiological profile and laboratory detection of extendedspectrum β-lactamases. Clinical ifectiouse diseases 42:153-163
CONCLUSION
• There is a high prevalence of ESBL-producing K. pneumonia
isolates in Pretoria Academic Hospital
• This leads to a need for improved, adequate infection control
measures to be instituted and more rational use of
antibiotics in Pretoria Academic Hospital