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.