SALMATcor: microagglutination for Salmonella flagella serotyping.

Salmonella is a complex bacterial group with more than 2400 serovars widely distributed in nature; they are considered zoonotic because they can infect a variety of animals and be transmitted to humans. Usually, they cause alimentary acquired diseases such as gastroenteritis, typhoid fever, and others that can lead to severe complications and death. Serotyping is useful to differentiate among Salmonella, because it shows an important correlation with their clinical and epidemiological patterns; consequently, it is of high value for public health, animal health, agriculture, and industry. To characterize all known Kauffmann-White Salmonella serovars, over 250 antisera are required. Due to this and to high prices antisera, many laboratories worldwide have limitations in establishing Salmonella surveillance. Therefore, we developed and validated a Salmonella flagella microagglutination test (SALMATcor) that significantly reduces laboratory requirements of antisera. SALMATcor is based on scaling down, by fivefold, the antigen:antiserum volumes actually required for the reference method: flagella standard tube agglutination technique (STAT). Antigen preparation, temperatures, and incubation periods remained as established for STAT. The SALMATcor was validated according to ISO/DIS 16140:1999 protocol, which included 1187 comparisons of flagella determinations conducted by SALMATcor and STAT, on 141 Salmonella isolates of 12 common serotypes and the use of antiserum recommended for STAT. SALMATcor concordance was excellent (Cohen's kappa index 0.9982), obtaining relative accuracy >99.9% and relative specificity >99.9%. Additionally, SALMATcor has been used by CNRB-INCIENSA since 2004 to respond to all 40 Salmonella proficiency testing strains, provided by World Health Organization-Global Salmonella Surveillance Network, obtaining 100% concordance on serovar identification. On the basis of the results achieved with SALMATcor and considering that it also significantly reduces antiserum expenses, hand labor, glassware, and bench top and water bath space requirements (microtiter plates and micropipette tips are the only additional supplies), we envision that SALMATcor will contribute to establish a sustainable Salmonella serovar surveillance worldwide.

Emergencia de β-lactamasa AmpC plasmídica del grupo CMY-2 en Shigella sonnei y Salmonella spp. en Costa Rica, 2003-2015 / Emergence of CMY-2-type plasmid-mediated AmpC β-lactamase in Shigella sonnei and Salmonella spp. in Costa Rica, 2003–2015

Las AmpC plasmídicas son enzimas del grupo de las β-lactamasas, codificadas por genes blaAmpC. Entre ellas, las del tipo CMY-2 son las que se reportan con mayor frecuencia a nivel mundial. La detección de enterobacterias productoras de AmpC plasmídicas CMY-2 es de importancia clínica, ya que pueden conducir a fracasos terapéuticos al emplear antibióticos b-lactámicos. Además, tienen importancia para la salud pública por su capacidad de transferirse por conjugación a otras enterobacterias, tanto en la comunidad como en ambiente nosocomial, por lo que se considera que tienen un claro potencial epidémico. Con el fin de conocer la circulación de este mecanismo de resistencia entre aislamientos de Salmonella y Shigella en Costa Rica, se realizó un análisis retrospectivo de la información contenida en las bases de datos de vigilancia de laboratorio del Centro Nacional de Referencia de Bacteriología (CNRB) del Instituto Costarricense de Investigación y Enseñanza en Nutrición y Salud (Inciensa), entre enero de 2003 y mayo de 2015. En dicho período se analizaron 4 363 aislamientos de Shigella y 1 785 aislamientos de Salmonella. Entre ellos se detectaron 15 aislamientos de Shigella sonnei y nueve de Salmonella (cuatro de origen clínico humano y cinco de origen aviar) con fenotipo sospechoso de portar AmpC plasmídica, todos los cuales se confirmaron pertenecientes al tipo CMY-2 mediante reacción en cadena de la polimerasa. Considerando estos resultados, se recomienda a los laboratorios de microbiología de la Red Nacional mantener la vigilancia y realizar la confirmación correspondiente de cualquier aislamiento sospechoso por métodos fenotípicos y moleculares. Lo anterior es especialmente importante en bacterias aisladas de infecciones extraintestinales, para evitar fallas en el tratamiento.

Plasmid-mediated AmpC are enzymes belonging to the group of β-lactamases and encoded by blaAmpC genes. Of these enzymes, those known as type CMY-2 are the most frequently reported worldwide. Detection of enterobacteria that produce CMY-2-type plasmid-mediated AmpC is clinically important since the use of β-lactam antibiotics can result in treatment failure. It is also important from a public health standpoint owing to the capacity for conjugative plasmid transfer to other enterobacteria, both within the community and in nosocomial environments. Thus, bacteria of this kind are considered to have clear epidemic potential. To investigate the circulation of this resistance mechanism among Salmonella and Shigella isolates in Costa Rica, from January 2003 to May 2015 we carried out a retrospective review of the data contained in the laboratory surveillance databases of the National Reference Bacteriology Center (CNRB) of the Costa Rican Nutrition and Health Research Institute (Inciensa). Over this period, 4363 Shigella isolates and 1785 Salmonella isolates were examined. Among them, 15 Shigella sonnei isolates and nine Salmonella isolates (four from human clinical specimens and five of avian origin) displayed a phenotype suspected of carrying plasmid-mediated AmpC. Polymerase chain reaction confirmed that all these isolates belong to type CMY-2. In light of these results, we recommend that the microbiology laboratories in the national network continue to conduct surveillance and confirm any suspicious isolates using phenotypic and molecular methods. This is particularly relevant when dealing with bacterial isolates from extraintestinal infections so as to prevent treatment failure.