Isolation, identification and characterization of Bordetella pertussis.

During the pertussis clinical trials, the best conditions for isolating B. pertussis were determined. Isolation is higher if two nasopharyngeal aspirates are collected, transport is carried out at ambient temperature and takes no longer than 48 hours, Reagan Lowe or Bordet Gengou selective and non-selective plates are used and incubated at 36 degrees C for seven days. Identification must include Gram stain and urease, oxidase and nitrate reactions. Confirmation is obtained with specific anti-B. pertussis antiserum. Characterization of the isolates can be performed by using the polymerase chain reaction, serotyping and pulsed-field gel electrophoresis.