Susana López Pineda Daniela Gallón Estrada

1. GENOTYPING OF ESBL
PRODUCING UROPATHOGENIC
Escherichia coli
IN WEST OF IRAN
SUSANA LÓPEZ PINEDA
DANIELA GALLÓN ESTRADA
MOLECULAR BIOLOGY
LINA MARTINEZ
III SEMESTER
PONTIFICIA BOLIVARIANA UNIVERSITY

2. INTRODUCTION
Escherichia
Escherichia is a genus of gram-negative,
non-spore
forming, facultative
anaerobic, rod-shaped
bacteria from the
family Enterobacteriaceae. In
those species which are
inhabitants of the gastrointestinal
tracts of warm-blooded
animals, Escherichia
provides a portion of the
production of vitamin K to its host.

3. INTRODUCTION
A number of the species
of Escherichia
are uropathogenic. The
genus is named
after Theodor Escherich
who
discovered Escherichia
coli.

4. INTRODUCTION
Although most species are harmless commensal
Escherichia some strains are human pathogens.
Some cause urinary tract infections and
gastrointestinal disease ranging from simple diarrhea
to dysentery. It is also responsible for a wide range of
disease states.

5. INTRODUCTION
Is a member of the
normal commensal
bowel flora of
humans and
colonization takes
place soon after
birth
E. coli

6. INTRODUCTION
Although Escherichia coli is
responsible for the majority
of disease in humans, other
members of the genus have
also been implicated in
human infections.

7. INTRODUCTION
It could have
pathogenic effects to
humans, because it can
live at intestines.
Although it is a normal
intestinal flora bacteria,
sometimes specific
strains have harmful
effects.
The diseases that result
from infection with E.coli
may be classified into two
groups:
Specific: Sign and
symptoms are related to
the site of colonization.
Non-specific: Signs and
symptoms are non related
to a site of mucosal
colonization.

8. INTRODUCTION
ESBL
B-LACTAMASES
Extended-Spectrum Beta-
Lactamases (ESBLs) are
enzymes that can be produced by
bacteria making them resistant to
cephalosporins e.g. cefuroxime,
cefotaxime and ceftazidime -
which are the most widely used
antibiotics in many hospitals

9. INTRODUCTION
ESBL infections
often affect the
urinary tract and
intestine, and
sometimes the
wounds and blood
are also infected

10. INTRODUCTION
B-Lactamases are
enzymes which
hydrolyze amide bond
of B-lactam ring .

11. INTRODUCTION
Antibiotic Resistance
Bacterial resistance is a
growing phenomenon
characterized by a partial
or total refractoriness of
microorganisms to the
antibiotic effect mainly due
to the indiscriminate and
irrational use of these.

12. INTRODUCTION
Bacterial antibiotic resistance
is acquired at times by
mutation of a gene that
modifies the structure of the
cell target.
Resistance may also be
acquired as a result of
bacterial cell infection by a
plasmid.

13. OBJECTIVE
The aim of the study
was to determine
genotyping of ESBL
producing UPECs

14. MATERIALES Y METODOS
Aislamientos
Personas con infección del tracto urogenital no
hospitalizadas
Oeste de Irán
Entre febrero de 2012 y febrero de
2013
Síntomas del tracto urinario inferior: Disuria, fiebre, frecuencia,
urgencia, y el crecimiento> 10 a la 5 colonias
Bioquímica y API20E

15. MATERIALES Y METODOS
API
Sistema de identificación multiprueba
Identificación rápida para bacterias de la
familia Enterobacteriaceae y otras bacterias Gram
21 Test bioquímicos estandarizados y miniaturizados, y una
base de datos.
Rápido, eficaz y permite realizar numerosas
pruebas a la vez.
20 Microtubos o pocillos con distintos sustratos
deshidratados.

16. MATERIALES Y METODOS
Susceptibilida
d
La meta principal del estudio de susceptibilidad es
proveer al clínico algunas recomendaciones sobre
la terapia que puede ser más apropiada en
pacientes con una infección específica.
Cefoperazona, ceftriaxona, cefuroxima, ciprofloxacina ,
piperacilina, gentamicina, amikacina , imipenem,
ampicilina, aztreonam, trimetoprim-sulfametoxazo,
carbenicilina , nitrofurantoína, y la ofloxacina.

17. MATERIALES Y METODOS
PCR
Amplificación enzimática de secuencias especificas
de ADN.
Polimeras
a
Síntesis Reparación Extiende o acorta ADN

18. MATERIALES Y METODOS
PFGE
Pulsed-Field Gel
Electrophoresis.
Electroforesis en gel de
campo pulsado.
Permite separar piezas
mucho más grandes de ADN
que la electroforesis en gel
de agarosa convencional.
PFGE se basa en la variable
de la migración de los
fragmentos de ADN grandes
en un campo eléctrico de

19. RESULTADOS
Figura 1

20. Figura 2
CHEF profiles of
UPEC strains
isolated. Lateral
lanes contain
Lambda Ladder
PFG Marker. Size
range: 48.5 kb to
679 kb (14
fragments).

21. Author Comment Yes or No
A.Davoodabadi,
A. Farahani, M.
Ranjbaran.(22)
UTIs are regarded as a health problem
around deferent
regions of the world. These isolates
are risk for public health such as in
both inpatient and outpatient specially
in some regions of Iran.
Yes
K. J. Sambrook,
D. W. Russel
(23)
PFGE analysis has been used in
epidemiological and molecular studies
of numerous bacterial and is gold
standard for molecular epidemiologic
in many bacteria such as E. coli
isolates.
This technique is better to identify the
source of infection and spread.
Yes
DISCUSSION

22. M. Anvarinejad,
Farshad Sh, R.
Ranjbar, G. M.
Giammanco,
A. Alborzi, A.
Japoni (24)
In study of Anvarinejad et al., the
lowest similarity was observed and
failed to find an association
between spread and colonization of
UTI isolates.
Yes

23. CONCLUSIONS
In this bacterial antibiotics
resistance era physicians
should know that now that is
not enough to know what is the
bacteria that is causing my
patient’s infection, we have to
deeper and use the different
tools that there are to figure
out what are the DNA
characteristics of this bacteria
and what could be the best
treatment for my patient.

24. Electrophoresis is a tool that
help us to know some DNA’s
bacterial characteristics and that
is a way figurate out why some
microorganisms are more
resistant than other ones.
Knowing about the virulence
factors of bacteria allow us to
fine other alternative
treatment’s sites and It could
impact in treatment of a
patient with and infection by a
resistance bacteria.
Urinary track infection is one of
the most common infections
around the world producing in
90% by E.coli bacteria.

27. BIBLIOGRAFÍA
• Sussman Max. Escherichia coli:
mechanisms of virulence. New York
• Electroforesis de proteínas y ácidos
nucleicos [sede web] Disponible en:
http://biomodel.uah.es/tecnicas/elfo/inicio.
htm