detection of clarithromycin-resistance helicobacter pylori strains in a dyspeptic patient population...
TRANSCRIPT
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Manuela Guzmán CastañedaJesús Sebastián Gómez
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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.
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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
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HELICOBACTER PYLORI
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HELICOBACTER PYLORI
BACTERIA RIBOSOME
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SYMPTOMS OF H. PYLORI INFECTION
Epigastric pain with burning
sensation.
Pain get worse with
empty stomach
Poor appetite
Weight loss Dyspepsia Vomiting Heart burn
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CLARITHROMYCIN
Clarithromycin binds to the subunit 23S
Bacteria ribosome
The bacteria cytoskeleton
collapse
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PCR METHOD
Polymerase Chain Reaction
Kary Mullis
Simple test to multiply DNA
Millions of copies
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PCR METHOD
Know partial sequence of the región of DNA
Enzymatic in vitro amplification
Increase the number of copies
Cycle 3 stages
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RFLP
Restriction Fragment Long Polymorphism
Restriction enzymes
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H. Pylori
Gastric ulcere
Epigastric pain
Illness
Clarithromycin
It binds to 23S
H. Pylori dies
Gastric mucose in perfect
conditions
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OBJECTIVE
“Investigate the proportion of common claritromycin-resistance mutation types present in the 23S rRNA gene of H. pylori strains in Sri Lanka.”
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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.
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METODOLOGÍA
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METODOLOGÍA
Endoscopia 2 Biopsias gástricas Prueba
de ureasa
Extracción de ADN
Amplificación con PCR glmM – 23S rRNA
Confirmación con histopatología
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MÉTODOS
Desnaturalización
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MÉTODOS
2. Alineamiento: glmM1-F - glmM2-R Cla18 - Cla21
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MÉTODOS
3.Síntesis
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MÉTODOS
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MÉTODOS
RFLP
MboII A2142G
Bsa I A2143G
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¿PARA QUE LA PCR?
Clonar fragmentos de ADN.
Detectar secuencias
Establecer polimorfismos
Rastreo de mutaciones.
Tipificación de DNA para trasplantes.
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¿PARA QUÉ LA RFLP?
Detectar mutaciones genéticas
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RESULTADOS
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DISCUSIÓN
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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
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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
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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.
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Jesús Sebastián Gómez
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Manuela Guzmán Castañeda
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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.