detection of clarithromycin-resistance helicobacter pylori strains in a dyspeptic patient population...
TRANSCRIPT
Manuela Guzmán CastañedaJesús Sebastián Gómez
INTRODUCTION The research article was realized in Sri Lanka, where has been reported a high prevalence of infection of Helicobacter Pylori. Scientist made a study to identify clarithromycin resistance of H. pylori strands.
Clarithromycin resistance is increasing, results in the use of empirical antibiotics for treatment.
HELICOBACTER PYLORI
Member of Campylobacter genus
Helicobacter genus 1989.
Gram-negative spiral-shape bacteria.
Infects up the 50% of human population.
Leading cause of peptic ulcers and gastriccancer.
Urease activity
HELICOBACTER PYLORI
HELICOBACTER PYLORI
BACTERIA RIBOSOME
SYMPTOMS OF H. PYLORI INFECTION
Epigastric pain with burning
sensation.
Pain get worse with
empty stomach
Poor appetite
Weight loss Dyspepsia Vomiting Heart burn
CLARITHROMYCIN
Clarithromycin binds to the subunit 23S
Bacteria ribosome
The bacteria cytoskeleton
collapse
PCR METHOD
Polymerase Chain Reaction
Kary Mullis
Simple test to multiply DNA
Millions of copies
PCR METHOD
Know partial sequence of the región of DNA
Enzymatic in vitro amplification
Increase the number of copies
Cycle 3 stages
RFLP
Restriction Fragment Long Polymorphism
Restriction enzymes
H. Pylori
Gastric ulcere
Epigastric pain
Illness
Clarithromycin
It binds to 23S
H. Pylori dies
Gastric mucose in perfect
conditions
OBJECTIVE
“Investigate the proportion of common claritromycin-resistance mutation types present in the 23S rRNA gene of H. pylori strains in Sri Lanka.”
METODOLOGÍA
Hospital de atención terciaria en Sri Lanka
Departamento de microbiología y de patología de la Universidad del estado de Sri Lanka.
Se realizó en 76 pacientes.
Aprobación ética por comité ético de la universidad del estado.
METODOLOGÍA
METODOLOGÍA
Endoscopia 2 Biopsias gástricas Prueba
de ureasa
Extracción de ADN
Amplificación con PCR glmM – 23S rRNA
Confirmación con histopatología
MÉTODOS
Desnaturalización
MÉTODOS
2. Alineamiento: glmM1-F - glmM2-R Cla18 - Cla21
MÉTODOS
3.Síntesis
MÉTODOS
MÉTODOS
RFLP
MboII A2142G
Bsa I A2143G
¿PARA QUE LA PCR?
Clonar fragmentos de ADN.
Detectar secuencias
Establecer polimorfismos
Rastreo de mutaciones.
Tipificación de DNA para trasplantes.
¿PARA QUÉ LA RFLP?
Detectar mutaciones genéticas
RESULTADOS
DISCUSIÓN
AUTHOR WHAT HE SAID ¿YES OR NO?RAYMOND J.(France)
Raymondet al.[20] in 2007 had found a 90% prevalence of A2143Gmutation
Yes
ABDOHALLI H.(Iran)
Abdollahi et al.,[18] in 2011 had shown a55% prevalence of A2142G mutation
Yes
AUTHOR WHAT HE SAID ¿YES OR NO?KIM KS. Kim KS
et al.[19] in 2002 had reported 100% mutation in A2143G orT2182C sites in Korea
Yes
FRANCESCVO VD. Published studies showedthat although A2142G mutation is present it sometimesmay not be phenotypically expressed.[14]
Yes
CONCLUSIONS
• Indiscriminated use of antibiotics increase emerging infections.
• Use of PCR is really important for the diagnostic of several patologies
• We concluded that the study of the molecular biology is now leading the investigations in Health sciences as a indispensable tool for the diagnosis and treatment of diseases.
• We recognize the importance of read and study in english even more in fields of science such as the Human Health knowing that the latest developments are published in this lenguaje.
Jesús Sebastián Gómez
Manuela Guzmán Castañeda
BIBLIOGRAFÍA
Yamamoto, Yoshimasa. Friedman, Herman. Hoffman, Paul. Helicobacter pylori Infection and Immunity.
David Martínez, Julían. Consuelo Henao, Sandra. Iván Lizarazo, Jorge. Resistencia antibiótica del Helicobacter pylori en América Latina y el Caribe. Revista Colombiana de gastroenterología. Vol.29 no.3 Bogotá Sept. 2014
Martínez Sánchez, Lina María. Vargad Grisales, Natalia. Osorio Ospina, Felipe. Ramírez Pulgarín, Sergio. Biología molecular. Octava edición. Universidad Pontificia Bolivariana.