Pertussis, or whooping cough, a highly contagious disease caused by Bordetella pertussis, is making a comeback globally and nationally in spite of reasonable vaccination coverage. This paper provides an update on laboratory testing methods that assist the confirmation of clinical disease and investigation of outbreaks. Laboratory confirmation of the diagnosis by polymerase chain reaction or serology should be attempted, especially when atypical pertussis is suspected clinically. Genetic and antigenic variations in virulence factors of strains circulating in the population should also be monitored.