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.

Infección por campylobacter y shigella como causa de diarrea aguda acuosa en niños menores de dos años en el distrito de La Victoria, Lima-Perú / Acute watery diarrhea caused by Campylobacter and shigella infection in children younger than two years old in La Victoria District, Lima-Peru

Objetivo: Determinar la frecuencia de Campylobacter y Shigella como agentes etiológicos en diarrea aguda acuosa en niños menores de dos años atendidos en 4 centros de salud del distrito de La Victoria (Lima, Perú). Materiales y métodos: Es este estudio transversal analítico se realizaron coprocultivos bajo técnica microbiológica estándar a los niños menores de dos años con diarrea aguda acuosa atendidos en 4 centros de salud de La Victoria entre abril y octubre de 2001. Fueron excluidos aquellos niños con disentería y aquellos que habían recibido tratamiento antibiótico previo a la atención. Resultados: se estudiaron un total de 248 casos, de los cuales 48 (19,4 por ciento) coprocultivos fueron positivos: 33 (13,3 por ciento) a Campylobacter, 12 (4,8 por ciento) a Shigella y 3 (1,2 por ciento) a Salmonella. No se identificaron otros patógenos. Las especies de shigella identificadas fueron: Shigella flexneri en 9 (75,0 por ciento) casos y Shigella sonnei en 3 (25,0 por ciento). La única especie de Campylocater y Shigella juegan un importante papel como agentes etiológicos causantes de diarrea aguda acuosa en niños menores de dos años.