ASSOCIATION OF EFFLUX PUMP GENES WITH

MULTI DRUG RESISTANCE OF KLEBSIELLA PNEUMONIA

IN NOSOCOMIAL INFECTIONS

Name of Student: Samman

L1F21MSMR0004
Supervisor: Dr. Saba Kabir
Co-supervisor: Dr. Farhan Rasheed
 Gram negative rod

INTRODUCTION lactose-fermenting

Oxidase negative

Non-motile

Facultative anaerobes

opportunistic human pathogen

Nosocomial infections
CONT…
Klebsiella infections commonly occur among sick patients
who are receiving treatment for other conditions.
Patients on ventilators or with intravenous catheters

Patients who are taking long courses of antibiotics

CONT..

(Gilles van Wezel 2018)

DISEASE CAUSED BY KLEBSIELLA

Urinary tract infection

Pneumonia

Septicemia

Soft tissue infections

ROLE OF EFFLUX PUMP

Efflux pumps are transport proteins involved in the

extrusion of toxic substances

Another factor that mediates antibiotic resistance

in bacteria is the possession of active efflux pumps

Efflux pumps are dominant in mediating resistance to

diverse classes of drugs and biofilm structures.
CONT..
Efflux pump systems expel a large array of chemical
compounds including antibiotics from bacteria cells
that possess them in an energy dependent manner

It not only extrude the given antibiotics out of the

bacterial cell but also act as virulence factors

adaptive responses that bestow antimicrobial resistance

during infections
(Kumar and Varela, 2013)
KNOWLEDGE GAP
In this era of antibiotic resistance, Klebsiella pneumoniae
represents one of the most concerning pathogens that causes
hospital acquired infection

According to WHO, multidrug resistant bacteria are emerging worldwide

which is a big challenge to healthcare

There is increasing evidence for the significant role of efflux pumps in

antibiotic resistance in bacteria
OBJECTIVE

To isolate and
Association of
identify Klebsiella To check the multi Molecular
efflux pump genes
pneumoniae from drug resistance of detection of efflux
with multi drug
different clinical Klebsiella pump genes in
resistance of
samples of pneumonia Klebsiella
Klebsiella
hospitalized isolates pneumonia
pneumoniae
patients
PROPOSED METHODOLOGY
• SAMPLE COLLECTION

pus swab urine

bronchial
blood catheters
washings

CSF
SAMPLE PROCESSING
Catalase
test TSI
With a straight inoculation needle,
touch the top of a well-isolated colony.

Inoculate TSI by first stabbing through the center of the

medium to the bottom of the tube.

Leave the cap on loosely and incubate the tube at 35°-37°C in

ambient air for 18 to 24 hours.
CONT…
citrate indole
Take a sterilized test tubes containing 4
Streak the slant back and forth with a ml of tryptophan broth. Inoculate the
light inoculum picked from the center tube aseptically by
of a well-isolated colony
taking the growth from 18 to 24 hrs culture.
Incubate the tube at 37°C for 24-28 hours.

Incubate aerobically at 35 to 37C for up to 4-7

days. Observe a color change from green to blue Add 0.5 ml of Kovac's reagent to the broth
along the slant. culture.

Observe for the presence or absence of ring

CONT..

DNA EXTRACTION

• DNA extraction will be carried out using Thermo scientific

DNA extraction kit™ as per manufacturer’s guidelines.
PCR
• The mmpl-7 and mmpl-11 primers will be used to determine
the efflux pump genes in Klebsiella pneumonia strains
REFERENCES
• Bergey, D. H. 1., and Holt, J. G. (2000). Bergey's manual of determinative bacteriology. 9th ed.
Philadelphia, Lippincott Williams & Wilkins)
• Davies J, Davies D. Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev
2010; 74 (03) 417-433
• Gniadkowski M. Evolution of extended-spectrum β-lactamases by mutation. Clin Microbiol
Infect 2008; 14 (Suppl. 01) 11-32
• Hou XH, Song XY, Ma XB, Zhang SY, Zhang JQ. Molecular characterization of multidrug-
resistant Klebsiella pneumoniae isolates. Braz J Microbiol 2015;46:759–68.