Aplicación del análisis de los polimorfismos de los tamaños de los fragmentos de restricción del gen flaA amplificado por reacción en cadena de la polimerasa y resistotipo para identificar potenciales brotes de campilobacteriosis no diagnosticados en España / Application of restriction fragment length polymorphism-polymerase chain reaction- flaA and resistotype to identify potential undiagnosed outbreaks of campylobacteriosis in Spain

INTRODUCCIÓN: Los brotes detectados de campilobacteriosis son poco frecuentes y, por lo general, cursan con un bajo número de pacientes, aunque se estima que muchos más permanecerían sin diagnosticar. Las técnicas de investigación de brotes más exitosas en Campylobacter spp. (PFGE, MLST) tienen el inconveniente de ser laboriosas y no estar disponibles en muchos laboratorios. MÉTODOS: Durante el año 2008 se recibieron 352 aislados de C. jejuni y C. coli procedentes de 16 hospitales. Todas las cepas fueron tipificadas genotípicamente mediante RFLP-PCR-flaA (tipo flaA) y fenotípicamente con su resistotipo. Se estableció que aquellas cepas de la misma especie procedentes del mismo hospital, aisladas en un periodo de hasta 11 días, con valores de CMI de±1 dilución y con el mismo tipo flaA, podrían pertenecer a un brote. Las cepas que cumpliesen con estos criterios serían posteriormente tipificadas mediante KpnI-PFGE y MLST. RESULTADOS: Veintitrés de los 352 aislados, en 10 grupos, cumplían con los criterios de pertenencia a posibles brotes no diagnosticados. En 8 grupos los pulsotipos (PFGE) de los aislados de cada grupo tenían una semejanza entre ellos mayor del 95%. En 7 de ellos los secuenciotipos (MLST) eran coincidentes. CONCLUSIONES: El uso de 2 marcadores sencillos (resistotipo y RFLP-PCR-flaA) puede detectar aislados que probablemente formen parte de un brote de campilobacteriosis no diagnosticado. Para su confirmación se requieren otros marcadores moleculares y los datos epidemiológicos de cada aislado. El estudio apunta a que, como en otros países, el número de brotes de campilobacteriosis en España está probablemente infraestimado

INTRODUCTION: Outbreaks of campylobacteriosis are infrequent and usually involve a low number of patients, although it is estimated that many more remain undiagnosed. The most successful techniques for outbreak investigation in Campylobacter spp. (PFGE, MLST) have the drawback of being laborious and not available in many laboratories. METHODS: During the year 2008, 352 isolates of C. jejuni and C. coli from 16 hospitals were received in our laboratory. All strains were genotyped by RFLP-PCR-flaA (flaA type) and phenotyped with their resistotype. It was established that the strains of the same species from the same hospital, isolated over a period of up to 11 days, with MIC values of±1 dilution with the same flaA type could belong to an outbreak. Strains that met these criteria would be later subtyped by KpnI-PFGE and MLST.RESULTS:A total of 23 out of 352 isolates, distributed in 10 groups, met the criteria for being associated with putative undiagnosed outbreaks. The similarity of the PFGE-profiles in 8 groups was greater than 95% among the isolates from each group. In 7 of the groups, the sequence types (MLST) were coincident. CONCLUSIONS: The use of 2 easy markers (resistotype and RFLP-PCR-flaA) may detect isolates probably belonging to an undiagnosed outbreak of campylobacteriosis. Accurate diagnosis requires other molecular markers and epidemiological data of each isolate. The study suggests that, as in other countries, the number of outbreaks of campylobacteriosis in Spain is probably underestimated

[Application of restriction fragment length polymorphism-polymerase chain reaction-flaA and resistotype to identify potential undiagnosed outbreaks of campylobacteriosis in Spain]. / Aplicación del análisis de los polimorfismos de los tamaños de los fragmentos de restricción del gen flaA amplificado por reacción en cadena de la polimerasa y resistotipo para identificar potenciales brotes de campilobacteriosis no diagnosticados en España.

INTRODUCTION: Outbreaks of campylobacteriosis are infrequent and usually involve a low number of patients, although it is estimated that many more remain undiagnosed. The most successful techniques for outbreak investigation in Campylobacter spp. (PFGE, MLST) have the drawback of being laborious and not available in many laboratories. METHODS: During the year 2008, 352 isolates of C. jejuni and C. coli from 16 hospitals were received in our laboratory. All strains were genotyped by RFLP-PCR-flaA (flaA type) and phenotyped with their resistotype. It was established that the strains of the same species from the same hospital, isolated over a period of up to 11 days, with MIC values of±1 dilution with the same flaA type could belong to an outbreak. Strains that met these criteria would be later subtyped by KpnI-PFGE and MLST. RESULTS: A total of 23 out of 352 isolates, distributed in 10 groups, met the criteria for being associated with putative undiagnosed outbreaks. The similarity of the PFGE-profiles in 8 groups was greater than 95% among the isolates from each group. In 7 of the groups, the sequence types (MLST) were coincident. CONCLUSIONS: The use of 2 easy markers (resistotype and RFLP-PCR-flaA) may detect isolates probably belonging to an undiagnosed outbreak of campylobacteriosis. Accurate diagnosis requires other molecular markers and epidemiological data of each isolate. The study suggests that, as in other countries, the number of outbreaks of campylobacteriosis in Spain is probably underestimated.

[Description of an outbreak of Campylobacter jejuni gastroenteritis and molecular characterization of the implicated strain]. / Descripción de un brote de gastroenteritis por y caracterización molecular de la cepa implicada.

INTRODUCTION: The aim of this study is to describe an outbreak of diarrhea caused by Campylobacter jejuni in a primary school. METHODS: Stool samples from five patients were cultured. Molecular typing of the isolated strains was performed using PCR-RFLP-flaA, PFGE and MLST. RESULTS: Campylobacter jejuni was isolated from all five patients. Two of the five strains were available for typing. The DNA patterns of the two isolates were indistinguishable. CONCLUSION: The results suggest that the causal strain had a common identity.

Descripción de un brote de gastroenteritis por y caracterización molecular de la cepa implicada / Description of an outbreak of Campylobacter jejuni gastroenteritis and molecular characterization of the implicated strain

Introducción. El objetivo de este trabajo es describir el estudio de un brote de diarrea por Campylobacter jejuni en una escuela infantil. Métodos. Se obtuvieron coprocultivos en 5 pacientes. La tipificación molecular se realizó mediante (RFLP-PCR flaA), electroforesis en campo pulsante (PFGE) y secuenciación de genes metabólicos (MLST). Resultados. Se aisló Campylobacter jejuni en los 5 enfermos. Sólo pudieron recuperarse para la tipificación 2 cepas. La tipificación mediante los 3 métodos resultó indistinguible en ambos aislamientos. Conclusión. Estos resultados sugieren una identidad común de la cepa causal (AU)

Introduction. The aim of this study is to describe an outbreak of diarrhea caused by Campylobacter jejuni in a primary school. Methods. Stool samples from five patients were cultured. Molecular typing of the isolated strains was performed using PCR-RFLP-flaA, PFGE and MLST. Results. Campylobacter jejuni was isolated from all five patients. Two of the five strains were available for typing. The DNA patterns of the two isolates were indistinguishable. Conclusion. The results suggest that the causal strain had a common identity (AU)