Resumen

QUINTERO ALVAREZ, Heidi  y  LEON ARCIA, Karen. Standard protocol for typing Pasteurella multocida by pulsed field electrophoresis in minigels. Rev. CENIC Cienc. Biol [online]. 2022, vol.53, n.1, pp. 69-79.  Epub 07-Mar-2022. ISSN 2221-2450.

Pasteurella multocida is a Gram-negative, bacillary bacterium that causes diseases of the upper respiratory tract in intensively farmed animals, causing great economic losses worldwide. Pulsed Field Electrophoresis (PFE) is the "gold standard" technique for molecular typing of strains involved in epidemiological outbreaks. However, a rapid standard protocol for typing P. multocida has not been reported. In the present work, the Pulse Time and Pulse Quantity parameters were standardized to separate the restriction fragments of the P. multocida genome in the minichambers of the Guefast06 System. Band patterns were obtained after 5h of electrophoresis, in contrast to 20-40h previously reported. These patterns obtained with Guefast06 were comparable to those reported in the literature because isoresolution of the DNA bands was achieved. Furthermore, it was determined that different strains of P. multocida can be discriminated when both the enzyme Apa I and Asc I are used to generate such band patterns. These experimental conditions could be advantageous for the investigation of infectious outbreaks since they would allow experts to make timely decisions in less time.

Palabras clave : Pulsed Field Electrophoresis; CHEF; Guefast; Pasteurella multocida.

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