Helicobacter pylori virulence genotypes in Bogotá River and wastewater treatment plants in Colombia.

BACKGROUND: Helicobacter pylori is a bacterium that infects 70%-80% of the population in Colombia, causing chronic gastritis in all those infected and gastric cancer in 1%-2% of those infected. In Colombia, some studies have identified the presence of vacA and cagA genes in environmental samples such as treated, surface, and wastewater, but they have not been evaluated in the Bogotá River. For this, the aim of this study was to identify the virulence genotypes of H. pylori present in samples from the Bogotá River and domestic wastewater treatment plants (WWTPs). MATERIALS AND METHODS: A total of 75 water samples (51 from the Bogotá River and 24 from wastewater treatment plants) were collected. The presence of H. pylori DNA and its virulence genotypes was determined by polymerase chain reaction (PCR). RESULTS: The presence of H. pylori DNA was demonstrated in 44% (33/75) of the samples, obtaining 63.6% (21/33) from the Bogotá River and 36.4% (12/33) from the WWTPs. The most prevalent H. pylori genotype was cagA (-) and vacAm1/s1/i1 being the most virulent of the vacA gene. CONCLUSIONS: This is the first study in Colombia that determines the cagA and vacA genotypes in surface water and WWTPs, indicating the circulation of virulent genotypes in the population. The presence of this pathogen in the waters can be represent a risk to the health of the surrounding population since these waters are reused by the communities for different purposes.

Bacteriophages of Helicobacter pylori.

The bacterium Helicobacter pylori colonize the stomach in approximately half of the world's population. Infection with this bacterium is associated with gastritis, peptic ulcer, adenocarcinoma, and gastric mucosa-associated lymphoid tissue lymphoma. Besides being a pathogen with worldwide prevalence, H. pylori show increasingly high antibiotic resistance rates, making the development of new therapeutic strategies against this bacterium challenging. Furthermore, H. pylori is a genetically diverse bacterium, which may be influenced by the presence of mobile genomic elements, including prophages. In this review, we analyze these issues and summarize various reports and findings related to phages and H. pylori, discussing the relationship between the presence of these elements and the genomic diversity, virulence, and fitness of this bacterium. We also analyze the state of the knowledge on the potential utility of bacteriophages as a therapeutic strategy for H. pylori.

Helicobacter pylori: Multiple resistance in patients from Bogotá, Colombia / Helicobacter pylori: resistencia múltiple en pacientes de Bogotá, Colombia

Introduction: The main cause for Helicobacter pylori infection treatment failure is antibiotic resistance, where clarithromycin and metronidazole play the main role. In Colombia, primary resistance as a consequence of the use of these two antibiotics and excessive levofloxacin use is above the accepted limit (13.6%, 83%, and 16%, respectively). Despite this fact, empirical therapies that include the combination of these antibiotics are used in patients with previous therapeutic failure. Objective: To determine antibiotic resistance in patients previously treated for H. pylori in Bogotá, Colombia. Materials and methods: We conducted a descriptive study that included ten isolates obtained from five patients with three or four previous failed treatments for H. pylori. Antibiotic resistance to amoxicillin, clarithromycin, levofloxacin, and metronidazole was investigated by agar dilution and confirmed by DNA sequencing (Magrogen, Korea). Results: Eight isolates were resistant to two or more antibiotics. All isolates were resistant to levofloxacin. Susceptibility patterns in isolates from the gastric antrum and the body of the stomach were different in three patients. Conclusion: As far as we know, this is the first evidence of multiple H. pylori resistance in Colombia in previously treated patients. Results demonstrated the consequences of using an ineffective antibiotic scheme and the need to assess antibiotic susceptibility in different anatomical sites of the stomach. The consequences of multiple resistance decrease possible antibiotic effectiveness to eradicate H. pylori in the future.

Introducción. La resistencia a los antibióticos es la principal causa del fracaso del tratamiento contra Helicobacter pylori; la claritromicina y el metronidazol son los antibióticos que generan mayor resistencia. En Colombia, la resistencia primaria a estos dos antibióticos y el uso excesivo de levofloxacina han alcanzado los límites aceptados (13,6, 83 y 16 %, respectivamente). A pesar de ello, se usa el tratamiento empírico combinando estos antibióticos en pacientes en los que ha fallado anteriormente. Objetivo. Determinar la resistencia a los antibióticos en pacientes previamente tratados para H. pylori en Bogotá, Colombia. Materiales y métodos. Se llevó a cabo un estudio descriptivo en el que se evaluó mediante dilución en agar la resistencia a la amoxicilina, la claritromicina, la levofloxacina y el metronidazol en 10 aislamientos provenientes de 5 pacientes con tres o cuatro tratamientos fallidos para H. pylori. La resistencia a los antibióticos se confirmó mediante secuenciación de ADN (Magrogen, Korea). Resultados. Ocho de los aislamientos presentaron resistencia a dos o más antibióticos y todos fueron resistentes a la levofloxacina. Los patrones de sensibilidad de los aislamientos provenientes del antro pilórico y del cuerpo del estómago, fueron diferentes en tres de los pacientes. Conclusión. Hasta donde se sabe, esta es la primera evidencia de resistencia múltiple de H. pylori en Colombia en pacientes previamente tratados. Los resultados evidenciaron las consecuencias del uso de un esquema ineficaz de tratamiento antibiótico y la necesidad de evaluar la sensibilidad a los antibióticos en diferentes sitios anatómicos del estómago. La resistencia múltiple limita el número de antibióticos útiles para erradicar H. pylori.

Correlation among fecal indicator bacteria and physicochemical parameters with the presence of Helicobacter pylori DNA in raw and drinking water from Bogotá, Colombia.

BACKGROUND: The quality of raw and drinking water is a matter of considerable concern due to the possibility of fecal contamination. To assess the quality and public health risk of different types of water, the fecal indicator bacteria (FIB) are used. However, some pathogens, such as Helicobacter pylori, may be present in water when FIB cannot be found. H pylori is recognized as the causative agent of chronic gastritis, peptic and duodenal ulcers, and gastric cancer. The aim of this study was to evaluate the relationships among physicochemical parameters, FIB concentrations, and the presence of H pylori DNA in raw and drinking water from Bogotá, Colombia. MATERIALS AND METHODS: A total of 310 water samples were collected 1 day per week from July 2015 to August 2016, and physicochemical parameters (pH, turbidity, conductivity, and residual free chlorine) were measured. Presence of H pylori DNA was determined and quantified by quantitative polymerase chain reaction (qPCR). Fecal indicator bacteria (total coliforms, Escherichia coli, and spores of sulfite-reducing Clostridia) were enumerated by using standard culture techniques. RESULTS: Thirty of 155 (31%) raw water samples and forty-eight of 155 (38.7%) drinking water samples were positive for the presence of H pylori. No statistically significant relationships were found between physicochemical parameters or FIB with the presence or absence of H pylori in any sample (P < 0.05). CONCLUSIONS: This study provides evidence of the presence of H pylori DNA in raw and drinking water in Bogotá, and shows that the detection and enumeration of FIB and physicochemical parameters in water do not correlate with the risk of contamination with H pylori.

Detection of Helicobacter pylori in drinking water treatment plants in Bogotá, Colombia, using cultural and molecular techniques.

Helicobacter pylori is one of the most common causes of chronic bacterial infection in humans, and a predisposing factor for peptic ulcer and gastric cancer. The infection has been consistently associated with lack of access to clean water and proper sanitation. H. pylori has been detected in surface water, wastewater and drinking water. However, its ability to survive in an infectious state in the environment is hindered because it rapidly loses its cultivability. The aim of this study was to determine the presence of cultivable and therefore viable H. pylori in influent and effluent water from drinking water treatment plants (DWTP). A total of 310 influent and effluent water samples were collected from three drinking water treatment plants located at Bogotá city, Colombia. Specific detection of H. pylori was achieved by culture, qPCR and FISH techniques. Fifty-six positive H. pylori cultures were obtained from the water samples. Characteristic colonies were covered by the growth of a large number of other bacteria present in the water samples, making isolation difficult to perform. Thus, the mixed cultures were submitted to Fluorescent in situ Hybridization (FISH) and qPCR analysis, followed by sequencing of the amplicons for confirmation. By qPCR, 77 water samples, both from the influent and the effluent, were positive for the presence of H. pylori. The results of our study demonstrate that viable H. pylori cells were present in both, influent and effluent water samples obtained from drinking water treatment plants in Bogotá and provide further evidence that contaminated water may act as a transmission vehicle for H. pylori. Moreover, FISH and qPCR methods result rapid and specific techniques to identify H. pylori from complex environmental samples such as influent water.

Eficacia de la terpia triple y cuadruple en helicobacter pylori y descripción de la resistencia a los principales antimicrobianos / Efficacy of triple and quadruple therapy in helicobacter pylori and description of resistance to antimicrobials

Helicobacter pylori es una bacteria gram negativa que infecta a más del 50 % de la población mundial. Su erradicación sigue siendo un reto, sobre todo por el aumento significativo de la prevalencia de la resistencia a los antibióticos como causa fundamental de fracaso terapéutico. Aunque el protocolo ideal de tratamiento no ha sido bien definido, se considera que el umbral de respuesta a la terapia de erradicación no debe ser inferior al 80 %. Es por esto que el presente estudio tiene como objetivo revisar literatura a nivel mundial desde el año 2010 al 2015, para evaluar la eficacia de la terapia triple y cuádruple para la erradicación del H. pylori en relación a la duración del tratamiento y la resistencia de la prevalencia a los principales antimicrobianos.

Helicobacter pylori is a gram-negative bacterium that directly infects more than 50 % of world population Eradicate it or diminish its increasing prevalence is still a challenge because its presence is still related with failed antibiotic treatments that create resistance. Despite there isn't an ideal protocol for its treatment, the threshold of response to eradication therapy should not be less than 80%. This study compilates world bibliographic information from 2010 to 2015; and it was made to determinate triple and quadruple therapy efficiency in H. pylori eradication, relating, treatment length, prevalence and resistance for the principal antimicrobial agents.

Frecuencia de mutaciones de la nitrorreductasa RdxA de Helicobacter pylori para la activación del metronidazol en una población del departamento del Cauca, Colombia / Frequency of Helicobacter pylori nitroreductase RdxA mutations for metronidazole activation in a population in the Cauca Department, Colombia

RESUMEN Introducción. La resistencia al metronidazol es un factor clave relacionado con el fracaso del tratamiento contra la infección por Helicobacter pylori asociada, principalmente, con mutaciones en la nitrorreductasa RdxA. A pesar de su importancia, los estudios sobre esta proteína son aún incipientes en Popayán, Colombia. Objetivo. Evaluar la frecuencia de las mutaciones en la nitrorreductasa RdxA en una población de pacientes con enfermedad gastrointestinal por H. pylori. Materiales y métodos. El ADN de 170 biopsias gástricas se amplificó mediante reacción en cadena de la polimerasa (PCR) para detectar las mutaciones en la nitrorreductasa RdxA. Se analizaron las secuencias traducidas a aminoácidos y se compararon con la cepa de referencia 26695. Resultados. La frecuencia de mutaciones de la nitrorreductasa RdxA en la población de estudio fue de 78 %. Su distribución más frecuente se detectó en las posiciones D59N (en 153 muestras), R131K (en 101 muestras), R90K (en 97 muestras), A118T (en 42 muestras), I160F (en 32 muestras), H97T (en 26 muestras) y en los codones de parada Q50*; D59*; E75*; C159* y I160* en cinco, una, tres, diez y seis muestras, respectivamente. El genotipo de virulencia más frecuente fue el vacAs1/m1 negativo para cagA (48,6 %). Conclusiones. La gran frecuencia de mutaciones en la nitrorreductasa RdxA en aislamientos de H. pylori en Popayán sugiere que los tratamientos empíricos con metronidazol no serían una opción válida para su erradicación en pacientes de la población estudiada.

ABSTRACT Introduction: Resistance to metronidazole is a key factor associated with Helicobacter pylori treatment failure. Even though resistance is mostly associated with RdxA nitroreductase mutations, studies of this H. pylori protein in Popayán (Colombia) are still incipient. Objective: To evaluate the frequency of mutations in the RdxA nitroreductase in a population of patients with H. pylori-positive gastrointestinal disease. Materials and methods: We amplified the DNA of 170 gastric biopsies by PCR to detect mutations in the RdxA nitroreductase. An analysis of DNA sequences translated into amino acid sequences was done and then compared to the reference strain 26695. Results: The frequency of RdxA nitroreductase mutations in this study population was 78%. Its most frequent distribution was found in positions D59N (153 samples), R131K (101 samples), R90K (97 samples), A118T (42 samples), I160F (32 samples) and H97T (26 samples), and meaningful stop codons Q50*, D59*; E75*, C159* and I160* in five, one, three, ten and six samples, respectively. The most common virulence genotype was vacAs1/m1 cagA negative (48.6 %). Conclusions: The high frequency of RdxA nitroreductase mutations in H. pylori isolates in Popayán (Colombia) indicates that empirical therapy with metronidazole may not be a valid option for the eradication of H. pylori in patients of the studied population.

[Frequency of Helicobacter pylori nitroreductase RdxA mutations for metronidazole activation in a population in the Cauca Department, Colombia].

INTRODUCTION: Resistance to metronidazole is a key factor associated with Helicobacter pylori treatment failure. Even though resistance is mostly associated with RdxA nitroreductase mutations, studies of this H. pylori protein in Popayán (Colombia) are still incipient. OBJECTIVE: To evaluate the frequency of mutations in the RdxA nitroreductase in a population of patients with H. pylori-positive gastrointestinal disease. MATERIALS AND METHODS: We amplified the DNA of 170 gastric biopsies by PCR to detect mutations in the RdxA nitroreductase. An analysis of DNA sequences translated into amino acid sequences was done and then compared to the reference strain 26695. RESULTS: The frequency of RdxA nitroreductase mutations in this study population was 78%. Its most frequent distribution was found in positions D59N (153 samples), R131K (101 samples), R90K (97 samples), A118T (42 samples), I160F (32 samples) and H97T (26 samples), and meaningful stop codons Q50*, D59*; E75*, C159* and I160* in five, one, three, ten and six samples, respectively. The most common virulence genotype was vacAs1/m1 cagA negative (48.6 %). CONCLUSIONS: The high frequency of RdxA nitroreductase mutations in H. pylori isolates in Popayán (Colombia) indicates that empirical therapy with metronidazole may not be a valid option for the eradication of H. pylori in patients of the studied population.

Inhibición del crecimiento de Erwinia chrysanthemi a diferentes concentraciones de ácido fólico: posible uso del ácido fólico como agente bacteriostático y fortificante de la papa Solanum tuberosum / Inhibition of Erwinia chrysanthemi growth to different concentrations of folic acid: possible use of folic acid as bacteriostatic agent and fortifying of Solanum tuberosum potato

Resumen Introducción: Las enterobacterias del genero Erwinia spp producen enfermedades en la papa, un tubérculo de consumo masivo. La regulación de la metilación del DNA puede regular la proliferación de la Erwinia, de tal modo que las concentraciones del ácido fólico, pueden tener un efecto en la capacidad patógena del microorganismo. De otra parte, el ácido fólico previene la aparición de defectos del tubo neural en humanos. Objetivo: Evaluar al ácido fólico como un agente bacteriostático de la Erwinia y que a su vez sea parte de la fortificación de alimentos de consumo masivo como la papa. Materiales y métodos: Se llevó a cabo la caracterización bioquímica de la Erwinia chrysanthemi, se estudió su crecimiento frente a diferentes concentraciones de ácido fólico Resultados: Al aumentar las concentraciones de la vitamina, desde 0,3 µg/L hasta 6,8 µg/L se inhibe el crecimiento bacteriano de la Erwinia chrysanthemi. La vitamina inhibe el crecimiento en cultivo de Erwinia chrysanthemi y actúa como como agente bacteriostático, aspecto es de gran relevancia dado que teóricamente, si la papa estuviera fortificada con el micronutriente, este actuaría contra el agente infeccioso y al mismo tiempo contribuiría al consumo adecuado de la vitamina en la población general.

Abstract Introduction: The enterobacteria of the Erwinia spp genus produce disease in potatoes, which is a tuber of mass consumption. The regulation of DNA methylation can regulate the proliferation of Erwinia in such a way that the concentrations of folic acid may have an effect on the microorganism pathogenic ability. On the other hand, the folic acid prevents the appearance of neural tube defects in humans. Objective: To evaluate folic acid as a bacteriostatic agent of Erwinia and, at the same time, as part of the fortification of mass consumption food such as the potatoes. Materials and methods: The biochemical characterization of the Erwinia chrysanthemi was carried out and its growth compared to different concentrations of folic acid was studied. Results: When increasing the concentrations of the vitamin from 0.3 µg/L up to 6.8 µg/L, the bacterial growth of Erwinia chrysanthemi is inhibited. The vitamin inhibits the growth in cultivation of Erwinia chrysanthemi and acts as a bacteriostatic agent. This aspect is of great importance given that, theoretically, if potatoes were fortified with micro-nutrient, this would act against the infectious agent and, at the same time, contribute to the adequate intake of the vitamin in the general population.

Helicobacter pylori y alteraciones endoscópicas en pacientes con obesidad severa, programados para cirugía bariátrica / Helicobacter pylori and Endoscopic Alterations in Severely Obese Patients Who Have Been Scheduled for Bariatric Surgery

En los pacientes con obesidad severa, la única intervención terapéutica eficaz es la cirugía bariátrica y la más frecuentemente utilizada es el bypass gástrico con reconstrucción en “Y” de Roux (BPGYR). Helicobacter pylori es la causa de la mayoría de úlceras pépticas y de cánceres gástricos. Muchos expertos recomiendan investigar y erradicar rutinariamente la infección antes del BPGYR. En nuestro medio no se ha investigado la prevalencia de H. pylori ni las alteraciones endoscópicas en pacientes programados para BPGYR, por lo cual se decidió realizar el presente trabajo. Materiales y métodos: se incluyeron pacientes adultos con obesidad severa, sin síntomas gastroduodenales, programados para BPGYR y sometidos a endoscopia digestiva alta preoperatoria. Resultados: desde marzo de 2007 a julio de 2014, se incluyeron 83 pacientes. El 87,95% eran mujeres. La edad promedio fue de 46,9±11,4 años (17-62 años). Se encontró H. pylori en el 57,83% (IC 95%: 47,09-66,96), similar a los no obesos. El 60% tenía algún grado de esofagitis erosiva, várices esofágicas en 2 pacientes y GIST en 1 paciente. Ninguno tuvo atrofia severa (OLGA III o IV). Conclusión: todos los pacientes tenían alguna alteración en la endoscopia. De estos, 2 pacientes tenían várices esofágicas. La prevalencia de H. pylori es similar a la de los no obesos. Se recomienda endoscopia alta de rutina en todos los pacientes antes BPGYR.

When the only effective therapeutic intervention for severely obese patients is bariatric surgery, the most frequently used method is a Roux-en-Y gastric bypass (RYGB). Helicobacter pylori (H. pylori) is the cause of most peptic ulcers and gastric cancers. Many experts recommend routine investigation and eradication of the infection before RYGB. The prevalence of H. pylori and endoscopic alterations among patients scheduled for RYGB had not been studied before in our environment, so we decided to conduct this research. Materials and methods: Severely obese adult patients with no gastric symptoms who were scheduled for RYGB and preoperative upper endoscopy were included in this study. Results: From March 2007 to July 2014, 83 patients were included. 87.95% were women. Mean age was 46.9 +/- 11.4 years (17-62 years). H. pylori was found in 57.83% (95% CI 47.09 - 66.96%) which is similar to the percentage of infections found in people who are not obese. 60% had some degree of erosive esophagitis including esophageal varices in two patients, and GIST in one patient. None had severe atrophy (OLGA III or IV). Conclusion: All patients had some alterations found during endoscopy. Two patients had esophageal varices. The prevalence of H. pylori is similar to that among people who are not obese. Routine upper endoscopy is recommended for all patients prior to RYGB.

[Determination of single nucleotide mutations in the 23S rRNA gene of Helicobacter pylori related to clarithromycin resistance in a population from Cauca, Colombia]. / Determinación de mutaciones de un solo nucleótido en el gen 23S rRNA de Helicobacter pylori relacionadas con resistencia a claritromicina en una población del departamento del Cauca, Colombia.

INTRODUCTION: Antibiotic combination therapy for the eradication of Helicobacter pylori should be based on local resistance patterns. OBJECTIVE: To d etermine the resistance of H. pylori to clarithromycin in a population from Cauca province, through the identification of mutations in the 23S rRNA gene in DNA from gastric biopsies. MATERIALS AND METHODS: A total of 162 patients with functional dyspepsia were included in the study. The 23S rRNA gene and the DNA from 162 gastric specimens were amplified by PCR, and the mutation pattern was identified by direct sequencing. RESULTS: The frequency of clarithromycin resistance was 4%. A2143G mutation was found in four patients (2.46%) and A2142G mutation was found in three patients (1.85%). CONCLUSIONS: Our study shows that the most frequent genotype in H. pylori -positive specimens was A2143G, followed by A2142G. The observed resistance prevalence of H. pylori was low; thus, we consider that clarithromycin treatment is a valid option for H. pylori eradication in the study population.

Determinación de mutaciones de un solo nucleótido en el gen 23S rRNA de Helicobacter pylori relacionadas con resistencia a claritromicina en una población del departamento del Cauca, Colombia / Determination of single nucleotide mutations in the 23S rRNA gene of Helicobacter pylori related to clarithromycin resistance in a population from Cauca, Colombia

Introducción. La terapia antibiótica combinada para la erradicación de Helicobacter pylori debería basarse en los patrones locales de resistencia. Objetivo. Determinar la resistencia de H. pylori a claritromicina en una población del departamento del Cauca mediante la identificación de mutaciones en el gen 23S r RNA en ADN obtenido de biopsias gástricas. Materiales y métodos. Se incluyeron en el estudio 162 pacientes con dispepsia funcional. El gen 23S rRNA se amplificó por PCR y el patrón de mutaciones se identificó por secuenciación directa. Resultados. La frecuencia de resistencia a claritromicina fue de 4 %. La mutación A2143G del gen se encontró en cuatro pacientes (2,46 %) y la mutación A2142G, en tres pacientes (1,85 %). Conclusiones. El estudio encontró que el genotipo más frecuente en los especímenes positivos para H. pylori fue 2143G, seguido por A2142G. La prevalencia observada de resistencia de H. pylori fue baja; por lo tanto, se considera que el tratamiento con claritromicina es una opción válida para la erradicación de H. pylori en la población objeto de estudio.

Introduction: Antibiotic combination therapy for the eradication of Helicobacter pylori should be based on local resistance patterns. Objective: To d etermine the resistance of H. pylori to clarithromycin in a population from Cauca province, through the identification of mutations in the 23S rRNA gene in DNA from gastric biopsies. Materials and methods: A total of 162 patients with functional dyspepsia were included in the study. The 23S rRNA gene and the DNA from 162 gastric specimens were amplified by PCR, and the mutation pattern was identified by direct sequencing. Results: The frequency of clarithromycin resistance was 4%. A2143G mutation was found in four patients (2.46%) and A2142G mutation was found in three patients (1.85%). Conclusions: Our study shows that the most frequent genotype in H. pylori -positive specimens was A2143G, followed by A2142G. The observed resistance prevalence of H. pylori was low; thus, we consider that clarithromycin treatment is a valid option for H. pylori eradication in the study population.

Resistencia de Helicobacter pylori a metronidazol, claritromicina y amoxicilina en pacientes colombianos / Helicobacter pylori resistance to metronidazole, clarithromycin and amoxicillin in Colombian patients

Helicobacter pylori (H. pylori) es un patógeno universal, que infecta a más de la mitad de la población mundial. En las últimas dos décadas, el tratamiento recomendado para su erradicación, como esquema de primera línea, es la triple terapia estándar, constituida por un inhibidor de la bomba de protones, amoxicilina y claritromicina o metronidazol. En los últimos años la eficacia de esta terapia ha declinado, debido especialmente a la resistencia de la bacteria a metronidazol y a claritromicina. Objetivos: En este estudio, se evaluó la prevalencia de resistencia primaria de cepas colombianas de H. pylori a metronidazol, claritromicina, amoxicilina. Además, se analizaron los genotipos de vacA y cagA de las cepas aisladas y la correlación entre los marcadores de virulencia y la resistencia a claritromicina, amoxicilina y metronidazol. Métodos: La resistencia a metronidazol, amoxicilina y claritromicina fue determinada por el método de E-test. Se extrajo el ADN genómico y variantes alélicas de vacA y cagA fueron identificadas por la técnica de reacción en cadena de la polimerasa (PCR). Resultados: La resistencia a metronidazol fue de 81,01% (IC 95% 70,3%-88,6%), a amoxicilina de 3,8% (IC 95% 0-8,6%) y a claritromicina de 17,72% (IC 95% 10,37-28,29). No se encontró asociación significativa entre el genotipo de patogenicidad y la resistencia o susceptibilidad a los antimicrobianos cuando los valores de CIM de cada antibiótico se compararon con los diferentes genotipos cagA y vacA. Conclusión: Encontramos una alta tasa de resistencia a los tres principales antibióticos utilizados en la mayoría de los esquemas exitosos de erradicación de la infección, lo cual implica la necesidad de investigar, con prioridad, nuevos esquemas de tratamiento para la erradicación de la infección en Colombia.

Helicobacter pylori (H. pylori), is a universal pathogen that infects more than half the world population. In the last two decades, the recommended treatment for its eradication, as first-line scheme is the standard triple therapy, consisting of an inhibitor of the proton pump, clarithromycin and amoxicillin or metronidazole. In recent years the effectiveness of this therapy has declined, especially due to the resistance of bacteria to metronidazole and clarithromycin. Objectives: In this study, we evaluated the prevalence of primary resistance of Colombian H. pylori isolates to metronidazole, clarithromycin, amoxicillin. In addition, the vacA and cagA genotypes of strains isolated were determined and associated to correlate the virulence markers and antibiotic resistance. Methods: Minimum inhibitory concentration (MIC) for metronidazole, clarithromycin and amoxicillin were determined by E-test method. Genomic DNA was extracted, and allelic variants of vacA and cagA were identified by the polymerase chain reaction (PCR). Results: Resistance to metronidazole was 81.01 % (IC95% 70.3%-88.6%), to amoxicillin 3,8% (IC 95% 0-8,6%), and to clarithromycin 17.72% (IC95% 10.37-28.29). No significant correlation between pathogenicity and resistance or susceptibility was detected when MIC values for each antibiotic were compared with different vacA and cagA genotypes. Conclusion: We find a high rate of resistance to three principal antibiotics used in the majority of the successful schemes of eradication of the infection, which implies the need to investigate with priority new schemes of treatment for the eradication of the infection in Colombia.

Comparación de la prueba de antígenos fecales (Elisa) y test de aliento de la urea frente a histología para el diagnóstico de Helicobacter pylori: Revisión sistemática de la literatura / Comparison of the enzyme immunoassay antigen test (Elisa) and urea breath test with histological methods for diagnosis of Helicobacter pylori infection: systematic literature review

El objetivo principal de este estudio fue realizar una revisión sistemática de la literatura para comparar la prueba de antígenos fecales y la prueba del aliento de la urea, frente a la histología en pre y postratamiento para la detección de Helicobacter pylori. La búsqueda de los artículos se realizó en las bases de datos PUBMED, SCIENCE DIRECT, OVID, COCHRANE y MEDICLATINA publicados entre 2003 y 2008. Los resultados de sensibilidad (S) y especificidad (E) de las pruebas en pretratamiento fueron: Antígenos fecales S: 98%, E: 95%; pruebas del aliento, S y E de 100% y en postratamiento ambas pruebas tuvieron S y E de 100%. Las gráficas de Funnel Plot en 3 de los 4 grupos revelaron asimetría, y el test de heterogeneidad mostró en los 4 grupos que los estudios eran homogéneos para S y E. En conclusión, la histología sigue siendo la mejor alternativa para el diagnóstico de la infección antes del tratamiento; por el contrario, las pruebas de aliento de la urea y antígenos fecales son las mejores opciones para verificar la erradicación de la infección, siendo la de antígenos fecales una que puede implementarse fácilmente en laboratorios de rutina en países en vías de desarrollo como Colombia, en donde no se realizan pruebas de forma rutinaria para verificar la erradicación de la infección.

This is a systematic review of literature of the results of sensibility (S) specificity (E), (VPP) positive predictive value, and (VPN) negative predictive value, of original articles published between 2003 and 2008, of the test of fecal antigens and urea breath test with the hematoxilin and eosin and giemsa stains in histology in pre and post treatment for the detection of Helicobacter pylori. The results showed S=98%, E 95% for the test of fecal antigens in pre treatment, E=100% y S=100% for the urea breath test of in pre treatment, E=100% y S=100% for the test of fecal antigens in post treatment and finally E=100% y S=100% for the urea breath test in post treatment. The graphs of Funnel Plot in 3 of the 4 groups revealed asymmetry, and the test of heterogeneity in the studies of the 4 groups were homogeneous so much for S as for E. In conclusion the histology continues being the best alternative for the diagnostic of the infection before the treatment; on the contrary the urea breath test and the test of fecal antigens are the best options to verify the eradication of the infection. The test of fecal antigens can be implemented easily in routine laboratories in developing countries as Colombia where there are not realized tests of the routine form to verify the eradication of the infection.

Comparación de las pruebas de dilución en Agar y PCR para determinación de susceptibilidad antimicrobiana de Helicobacter pylori. Revisión sistemática de la literatura / Comparation of Agar dilution test with Polymerase Chain Reaction (PCR) to determinate antimicrobial susceptiblility of Helicobacter Pylori. A systematic literature review

Objetivo: determinar las características operativas de la prueba RFLP-PCR frente a la prueba dilución en agar para evaluar la susceptibilidad antimicrobiana a claritromicina en aislamientos clínicos de H. pylori. Metodología: la búsqueda de estudios de pruebas diagnósticas sobre resistencia antimicrobiana de H. pylori a claritromicina con técnicas de dilución en agar y RFLP-PCR se realizó en Medline, Science direct, Ovid y Cochrane. Se elaboraron tablas de contingencia para calcular las características operativas, en el programa RevMan 5. La heterogeneidad fue evaluada por la gráfica Forest Plot y el estadístico de Q. La presencia de sesgos de publicación se evaluó con Funnel Plot. Resultados: doce artículos cumplieron con los criterios de inclusión. El overall de especificidad fue 100% (IC 95% 91-100), demostrando baja probabilidad de falsos positivos. Para sensibilidad el valor fue de 91% (IC 95% 88-94) indicando una mayor probabilidad de resultados falsamente negativos. En la gráfica de “Funnel Plot” se observó asimetría para ambas características demostrando sesgo de publicación. Conclusiones: la técnica RFLP-PCR no presentó características operativas iguales o superiores al 95%, comparada con el estándar de referencia dilución en agar. Por lo anterior esta técnica de se debe considerar la prueba de elección cuando se estudie la susceptibilidad antimicrobiana de H. pylori.

Aim: Establish the available scientific evidence of the operational characteristics of PCR-RFLP test with Agar Dilution for the determination of antimicrobial susceptibility in clarithromycin clinical isolates of Helicobacter pylori. Methods: We have performed the search bibliography about diagnostic tests on antimicrobial resistance of H. pylori to clarithromycin by using Agar Dilution and PCR-RFLP techniques over Medline, Science direct, Ovid and Cochrane of studies. The information was validated by two observers who checked the inclusion criteria and quality. We obtained the operational characteristics of the studies on contingency tables; analysis was performed on the RevMan 5 program. Results: A total of 50 references were tested from those 12 has been chosen in accord with the inclusion criteria and analyzed as summary measures. The specificity “overall” was a 100 % (CI 95% 91-100) that demonstrate a low probability of positive false. The projected overall sensitivity was 91% (CI 95% 88-94%) which indicated a high probability of negative results. The test showed heterogeneity studies homogeneous in sensitivity and specificity (p = 0.78) (p = 0.99). The graphics of “Funnel Plot” revealed asymmetry for both of those characteristics that showed a publications bias. In the group analysis have not found antibiotics different from clarithromycin and was evident that the continent was more publications Europe, followed by Asia and Latin America. Conclusion: The sensitivity and specificity of PCR-RFLP technique for clarithromycin not have values equal to or higher than 95% compared proof Agar Dilution.

Immune responses to isolated human skin antigens in actinic prurigo.

BACKGROUND: Actinic prurigo (AP) is a frequent photodermatosis among Amerindians, with a high incidence among women and children below ten years of age. Neither the cause of actinic prurigo nor its etiological agent have been described. Not much is known about the pathogenic mechanisms of the disease, although associations with the human leucocitary antigens (HLA) and local immune responses seem to play an important role in its expression, as is the case in other skin autoimmune disorders, such as pemphigus and psoriasis. MATERIAL/METHODS: In this paper we compare cellular and humoral immunity through in vitro proliferation studies, ELISA and immunofluorescence tests in actinic prurigo patients and healthy controls. RESULTS: Autoantibody reactivities on the skin and also proliferative responses to isolated autologous skin antigens were higher in patients than in controls. The polyclonal cellular immune response against T cell mitogens and against allogeneic stimuli was found to be diminished in patients. CONCLUSIONS: We found autoimmune reactivity in patients suffering from actinic prurigo. We postulate that AP patients may have one or more skin antigens that stimulate an autoimmune response, which causes the observed skin lesions. As AP is a pathology that affects mainly the skin, any immune response should be localized and the observed infiltrating lymphocytes in skin biopsies should be activated by these hypothetical antigens.

Virus diversity of acute diarrhea in tropical highlands.

Infectious acute diarrhea (IAD) is an important health problem affecting a large number of Latin-American children. Several reports show that bacteria, parasites and virus are involved in the burden of this disease. Most reports reveal Rotavirus A as the responsible etiological agent, at the same time, there seems to be some correlation between IAD and seasonal weather changes. To learn about the type of microbial agents associated with IAD in children during mildly changing yearly climatic conditions, as found in a high altitude tropical city, and to identify the viral agents affecting this population, stool samples from 300 children under 5 years of age were studied throughout a one-year period. Bacteria and intestinal parasites were identified by routine methods, while viruses were detected and typed by EIA and PCR. 20.6% of the IAD studied was associated with bacteria; 9% with parasites and 40% with virus. Group C Rotavirus accounted for 20.2%, group A Rotavirus for 13% and Calicivirus 10%. During November-April (p < 0.007) more virus associated IAD was found, while bacteria (p < 0.03) or parasite (p < 0.00014) related IAD was prevalent from May to October. The mild seasonal weather changes don't seem to be associated with any other microbial agent.

Comportamiento de la diarrea causada por virus y bacterias en regiones cercanas a la zona ecuatorial / Behavior of diarrhea caused by viruses and bacterias in regions located near the Equator zone

INTRODUCCIÓN: La enfermedad diarreica aguda (EDA) es un problema de salud a nivel mundial que afecta a la población infantil de distintas regiones. Casi todos los estudios epidemiológicos se han hecho en países con estaciones y poco se informa su comportamiento en países sin estaciones, donde la EDA es endémica con picos epidémicos. OBJETIVOS: Contribuir a conocer la conducta de EDA en Colombia y determinar si su comportamiento es diferente en niños menores de cinco años en dos regiones distintas entre sí en geografía y clima. MATERIALES Y MÉTODOS: Se hizo un estudio descriptivo en dos localidades colombianas. Una en la costa atlántica y otra en el centro del país. La muestra se obtuvo en menores de cinco años que consultaron por diarrea a centros asistenciales de cada región. Los microorganismos bacterianos se identificaron mediante pruebas bioquímicas y los virus con técnicas inmunoenzimáticas. En el análisis estadístico se siguieron un ensayo bivariado y pruebas Z de normalidad para verificar si el clima modifica el comportamiento de EDA y si se presenta de manera distinta en las dos regiones. RESULTADOS: En ambas zonas (Cartagena, Bolívar y Facatativa, Cundinamarca) predominó la diarrea viral, frente a la EDA bacteriana. También en ambas el rotavirus fue prevalente. Fue mucho más baja la presencia de astrovirus y adenovirus. No hubo datos con significación estadística para demostrar que las condiciones ambientales y las propias de los niños, alteran el comportamiento de la EDA, pero sí se observó que la EDA por rotavirus se comporta de manera diferente al analizar en forma comparativa las dos regiones del estudio