seminario biologia molecular syd
TRANSCRIPT
GENOTYPING OF ESBL PRODUCING UROPATHOGENIC Escherichia coli
IN WEST OF IRAN
SUSANA LÓPEZ PINEDADANIELA GALLÓN ESTRADA
MOLECULAR BIOLOGYLINA MARTINEZ
III SEMESTERPONTIFICIA BOLIVARIANA UNIVERSITY
INTRODUCTION
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.
Escherichia
A number of the species of Escherichia are uropathogenic. The genus is named after Theodor Escherich who discovered Escherichia coli.
INTRODUCTION
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.
Is a member of the normal commensal bowel flora of humans and colonization takes place soon after birth
INTRODUCTION
E. coli
Although Escherichia coli is responsible for the majority of disease in humans, other members of the genus have also been implicated in human infections.
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.
INTRODUCTION
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.
ESBLB-LACTAMASES
INTRODUCTION
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
ESBL infections often affect the urinary tract and intestine, and sometimes the wounds and blood are also infected
INTRODUCTION
INTRODUCTION
B-Lactamases are enzymes which hydrolyze amide bond of B-lactam ring .
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.
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.
The aim of the study was to determine genotyping of ESBL producing UPECs
OBJECTIVE
MATERIALES Y METODOS
Aislamientos
Personas con infección del tracto urogenital no hospitalizadas
Oeste de Irán
Síntomas del tracto urinario inferior: Disuria, fiebre, frecuencia, urgencia, y el crecimiento> 10 a la 5 colonias
Entre febrero de 2012 y febrero de 2013
Bioquímica y API20E
MATERIALES Y METODOS
API
Sistema de identificación multiprueba
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.
Identificación rápida para bacterias de la familia Enterobacteriaceae y otras bacterias Gram
Susceptibilidad
MATERIALES Y METODOS
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.
MATERIALES Y METODOS
PCR
Amplificación enzimática de secuencias especificas de ADN.
Polimerasa
Síntesis Reparación Extiende o acorta ADN
PFGE
MATERIALES Y METODOS
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 polaridad alterna.
Figura 1
RESULTADOS
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).
Author Comment Yes or No
A.Davoodabadi, A. Farahani, M. Ranjbaran.(22)
UTIs are regarded as a health problem around deferentregions 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
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
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.
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.
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