Validation of the T86I mutation in the gyrA gene as a highly reliable real time PCR target to detect Fluoroquinolone-resistant Campylobacter jejuni.

BACKGROUND: Campylobacter jejuni is a leading cause of bacterial diarrhea worldwide, and increasing rates of fluoroquinolone (FQ) resistance in C. jejuni are a major public health concern. The rapid detection and tracking of FQ resistance are critical needs in developing countries, as these antimicrobials are widely used against C. jejuni infections. Detection of point mutations at T86I in the gyrA gene by real-time polymerase chain reaction (RT-PCR) is a rapid detection tool that may improve FQ resistance tracking. METHODS: C. jejuni isolates obtained from children with diarrhea in Peru were tested by RT-PCR to detect point mutations at T86I in gyrA. Further confirmation was performed by sequencing of the gyrA gene. RESULTS: We detected point mutations at T86I in the gyrA gene in 100% (141/141) of C. jejuni clinical isolates that were previously confirmed as ciprofloxacin-resistant by E-test. No mutations were detected at T86I in gyrA in any ciprofloxacin-sensitive isolates. CONCLUSIONS: Detection of T86I mutations in C. jejuni is a rapid, sensitive, and specific method to identify fluoroquinolone resistance in Peru. This detection approach could be broadly employed in epidemiologic surveillance, therefore reducing time and cost in regions with limited resources.

Campylobacter antimicrobial resistance in Peru: a ten-year observational study.

BACKGROUND: Campylobacter jejuni and Campylobacter coli are food-borne pathogens of great importance and feature prominently in the etiology of developing world enteritis and travellers' diarrhoea. Increasing antimicrobial resistant Campylobacter prevalence has been described globally, yet data from Peru is limited. Our objective was to describe the prevalence trends of fluoroquinolone and macrolide-resistant C. jejuni and C. coli stool isolates from three regions in Peru over a ten-year period. METHODS: Surveillance for enteric pathogens was conducted in Lima, Iquitos and Cusco between 2001 and 2010. Campylobacter stool isolates were tested for susceptibilities to ciprofloxacin, azithromycin and erythromycin. Susceptibilities were reviewed for 4652 isolates from Lima ( n = 3419), Iquitos ( n = 625) and Cusco ( n = 608). RESULTS: Comparing the study periods of 2001-2005 and 2006-2010, prevalence of ciprofloxacin-resistant C. jejuni isolates rose in the study areas of Lima (73.1% to 89.8%, p < 0.001) and Iquitos (24.1% to 48.9%, p < 0.001). Ciprofloxacin-resistant C. coli rates also increased in Lima (48.1% to 87.4%, p < 0.001) and Cusco (10.0% to 65.9%, p = 0.005). Small but significant increases in azithromycin-resistant and erythromycin-resistant C. jejuni prevalence were noted in Iquitos (2.2% to 14.9%, p < 0.001; 3.2% to 14.9%, p = 0.002), and erythromycin-resistant C. coli rates increased in Lima (0.0% to 5.3%, p = 0.038). The prevalence of C. jejuni isolates resistant to both ciprofloxacin and azithromycin increased in Iquitos (0.3% to 14.9%, p < 0.001) and Lima (0.3% to 1.6%, p = 0.011), and prevalence of C. jejuni isolates resistant to both ciprofloxacin and erythromycin rose in Iquitos (0.0% to 14.9%, p < 0.001). Ciprofloxacin and erythromycin resistant C. coli prevalence increased in Lima (0.0% to 5.3%, p = 0.034). CONCLUSIONS: These results have implications for the empirical management of enterocolitis in Peru. Ongoing surveillance is essential to guide appropriate antimicrobial use in this setting. Local epidemiological studies to explore the relationship between increasing antimicrobial resistance and agricultural or human antibiotic use may be valuable.

BASAN index (Body mass index, Age, Sex, Arterial hypertension and Neck circumference) predicts severe apnoea in adults living at high altitude.

OBJECTIVES: Obstructive sleep apnoea (OSA)/hypopnoea syndrome is associated with serious and major multiorgan morbidities, particularly in its most severe forms. However, no severe OSA screening instruments are available for high altitude residents that enable adequate identification and clinical prioritisation of such patients. We aimed at developing a severe OSA prediction tool based on the clinical characteristics and anthropometric measurements of a clinical referral cohort living at 2640 m.a.s.l. DESIGN: Cohort-nested cross-sectional study. SETTING: Sleep laboratory for standard polysomnography (PSG) in Colombia. PARTICIPANTS: A predictive model was generated from 8718 participants referred to the PSG laboratory. Results were subsequently validated in a second cohort of 1898 participants. PRIMARY OUTCOME: To identify clinical and anthropometric variables associated with severe OSA (>30 events/hour) and to include them in a binary logistic regression model. RESULTS: The significant variables that were retained with the presence of severe OSA included Body mass index (BMI), Age, Sex, Arterial hypertension and Neck circumference (BASAN). The area under the receiver operating characteristic curvefor the BASAN index was 0.69 (95% CI: 0.68 to 0.70) in the derivation cohort and 0.67 (95% CI: 0.65 to 0.69) in the validation cohort, whereby a BASAN index ≥2 had a sensitivity of 95% and a specificity of 17% to detect severe OSA. CONCLUSION: An objectively based approach to screen for the presence of severe OSA, the BASAN index, exhibits favourable sensitivity characteristics that should enable its operational use as a screening tool in a Hispanic population with a clinical suspicion of OSA and living at high altitude.

ß-lactamasas de espectro extendido tipo CTX-M en aislamientos clínicos de Escherichia coli y Klebsiella pneumoniae en el Instituto Nacional de Salud del Niño-Breña, Lima, Perú / Extended spectrum type CTX-M ß-lactamases in clinical infections caused by Escherichia coli y Klebsiella pneumoniae at the Instituto Nacional de Salud del Niño-Breña, Lima, Peru

Objetivo: Determinar la frecuencia de β-lactamasas de espectro extendido (BLEE) en Escherichia coli y Klebsiella pneumoniae y la frecuencia de CTX-M en las productoras de BLEE en el Instituto Nacional de Salud del Niño - Breña (INSN-B). Material y métodos: Se analizaron enterobacterias productoras de BLEE del INSN-B entre los meses de agosto de 2012 y enero del 2013. Se incluyeron 724 aislamientos de Escherichia coli y 181 aislamientos de Klebsiella pneumoniae, consecutivos no repetidos, de pacientes hospitalizados y de la comunidad. La identificación se realizó por bioquímica convencional. la detección fenotípica de BLEE se hizo por el método de Jarlier y la detección genotípica de CTX-M mediante reacción en cadena de la polimerasa (PCR). Resultados: 281 (31%) de los aislamientos de ambas enterobacterias fueron productoras de BLEE; 207/724 (28,6%) E. coli y 74/181 (40,9%) K. pneumoniae. Se detectó el gen bla en 256 de los aislamientos productores de BLEE (91,1%). Conclusiones: Las BLEE de tipo CTX-M están presentes en nuestra institución, a pesar que nuestros datos representan una sola institución, brinda parte del panorama nacional sobre la resistencia a los antimicrobianos; por lo tanto, el enfoque de epidemiológico molecular es importante para desarrollar más y mejores estrategias de control y manejo de estos patógenos en nuestro país.

Objective: To determine the frequency of extended spectrum β-lactamases (ESBL) in clinical infections caused by Escherichia coli y Klebsiella pneumoniae and the frequency of CTX-M among them at the Instituto Nacional de Salud del Niño-Breña, Lima, Peru. Methods: ESBL producing strains of E. coli and K. pneumoniae collected from August 2012 and January 2013 were analyzed; a total of 724 E. coli and 181 K. pneumoniae consecutive, non- repeated isolates from community and hospital acquired infections were included. Identification was performed by conventional biochemistry, ESBL phenotype was detected following the Jarlier´s method and PCR was used to detect CTX-M. Results: overall prevalence of ESBL was 31% (281 strains); 207/724 (28.6%) E. coli and 74/181 (40.9%) K. pneumoniae. The bla gene was detected in 256 of ESBL producing strains (91.1%). Conclusions: The CTX-M phenotype of ESBL producing strains is present in our institution. Despite of showing information of a single institution, these data bring a glance of what the antimicrobial resistance pattern may be at a national level and underscores the utility of molecular biology in designing preventing measures.

[Lophomonas sp. in respiratory tract secretions in hospitalized children with severe lung disease]. / Hallazgo de Lophomonas sp. en secreciones del tracto respiratorio de niños hospitalizados con enfermedad pulmonar grave.

Lophomonas sp. is a habitual parasite of the intestinal tract of the cockroaches and that is not recognized as pathogenic human being. Nevertheless, in the world literature are few reports of Lophomonas sp. in respiratory tract secretions in patients with severe pulmonary disease, mostly in adults. We present evidences of Lophomonas sp. in the respiratory low tract of children attended in the national reference center of paediatric diseases of Lima, Peru, in the period 2009- 2010. We found six cases, 4/23 from broncoalveolar lavage and 2/794 from tracheal aspirate samples of children. Five of them had pneumonia and one atelectasis, four were hospitalized in the intensive care unit. Increase knowledge about the presence of this organism in respiratory infections is needed, as its real pathogenic role.

Indicación de antibióticos en niños con faringoamigdalitis aguda y test positivo para estreptococo beta hemolítico del grupo A atendidos en el Instituto Nacional de Salud del Niño, Lima – Perú / Indication of antibiotics for children with pharyngitis acute and positive test for beta hemolytic strepotococcus group A treated at the National Institute of Child Health, Lima – Per£

Objetivo: Conocer la frecuencia de la indicación de antibióticos en niños con diagnóstico clínico de faringoamigdalitis aguda (FAA) y test rápido positivo para estreptococo beta hemolítico del grupo A atendidos en consultorios externos de pediatría, y el tipo de antibiótico indicado con más frecuencia. Material y Métodos: Estudio descriptivo, transversal. La muestra fue de 351 niños de 3 a 15 años diagnosticados como FAA. Abordados a la salida de la consulta y con firma y consentimiento informado a quienes se les aplico una encuesta y se realizó toma de muestra de secreciones faringoamigdalianas, las fueron analizadas mediante el test de inmunoaglutinación para EbhGA (ACON) con una S=91% y E=98%. Resultados: la frecuencia de prescripción antibiótica fue de 42.7% (150). El antibiótico más usado fue la penicilina con 25.4 %, seguido de Macrólidos (14.2%). 15 casos (4.3%) de faringoamiggalitis bacteriana tuvieron Test EbhGA positivo. Nueve de ellos (60%) recibieron antibióticos, la inflamación y el exudado amigdalino se presentaron en 320 (93,4%). Hubo ausencia de tos en 46 casos (13,1%). Se indicó sintomáticos en 51.5% de pacientes. Conclusiones: La frecuencia de FA estereocoptocócica fue baja según el test de detección rápida, la prescripción antibiótica fue 10 veces más indicada que la positividad del test; el antibiótico más utilizado fue la penicilina.

Objective: Generally is to determine the frequency of indication of antibiotic for children with clinically diagnosed acute pharyngitis and positive test for beta hemolytic streptococcus Group A treated at pediatric, clinics, a specific objective is to determine the frequency with which it is negative test for the detection of beta-hemolytic streptococcus for type A (GABHS) in these patients. Material and Methods: We made across-sectional study. The sample was 351 children of 3-15 years diagnosed by his physician as FAA, the departure of your inquiry, the informed consent, they have participated in a survey of the study variables and underwent sampling pharyngeal secretions, these were analyzed using test for GABHS inmunoagglutination (ACON) with S=91%, E=98%. Results: The frequency of antibiotic prescriptions was 42.7% (150). The first common antibiotic was penicillin with 25.4%, the second one, macrolides with 14.2%. There was a 4.3% (15) case of GABHS bacteria pharyngeal confirmed by positive test. Inflammation and exudate tonsillar was presented in 93.4% (328). There was lack of cough in 13.1% (46). The other antibiotic was symptomatic indication in 51.5% of patients. Conclusions: The antibiotic prescription rate more than the actual by the positive test; the most common antibiotic was penicillin. The test was positive in percentage and the mostly is in the school group, the most common symptom was swelling and amygdaline exudate. The other symptomatic medications were given.

Hallazgo de Lophomonas sp. en secreciones del tracto respiratorio de niños hospitalizados con enfermedad pulmonar grave / Lophomonas sp. in respiratory tract secretions in hospitalized children with severe lung disease

Lophomonas sp. es un parásito habitual del tracto intestinal de las cucarachas y que no es reconocido como patógeno humano. Sin embargo, en la literatura mundial existen escasos reportes de Lophomonas sp. en secreciones del tracto respiratorio en pacientes con enfermedad pulmonar grave, principalmente en adultos. Presentamos evidencias de Lophomonas sp. en el tracto respiratorio inferior de niños atendidos en el centro de referencia nacional de enfermedades pediátricas de Lima, Perú, en el periodo 2009-2010. Se encontró seis casos, 4/23 provenientes de muestras de lavado broncoalveolar y 2/794 muestras de aspirado traqueal de niños. Cinco de ellos tuvieron neumonía y uno atelectasia, cuatro estuvieron en la unidad de cuidados intensivos. Es necesario conocer más sobre la presencia de este organismo en infecciones respiratorias así como su rol patogénico real.

Lophomonas sp. is a habitual parasite of the intestinal tract of the cockroaches and that is not recognized as pathogenic human being. Nevertheless, in the world literature are few reports of Lophomonas sp. in respiratory tract secretions in patients with severe pulmonary disease, mostly in adults. We present evidences of Lophomonas sp. in the respiratory low tract of children attended in the national reference center of paediatric diseases of Lima, Peru, in the period 2009- 2010. We found six cases, 4/23 from broncoalveolar lavage and 2/794 from tracheal aspirate samples of children. Five of them had pneumonia and one atelectasis, four were hospitalized in the intensive care unit. Increase knowledge about the presence of this organism in respiratory infections is needed, as its real pathogenic role.