Prevalence and genetic characterization of pertactin-deficient bordetella pertussis in Japan

The adhesin pertactin (Prn) is one of the major virulence factors of Bordetella pertussis, the etiological agent of whooping cough. However, a significant prevalence of Prn-deficient (Prn ^-) B. pertussis was observed in Japan. The Prn ^- isolate was first discovered in 1997, and 33 (27%) Prn ^- isolates were identified among 121 B. pertussis isolates collected from 1990 to 2009. Sequence analysis revealed that all the Prn ^- isolates harbor exclusively the vaccine-type prn1 allele and that loss of Prn expression is caused by 2 different mutations: an 84-bp deletion of the prn signal sequence (prn1ΔSS, n = 24) and an IS481 insertion in prn1 (prn1::IS481, n = 9). The frequency of Prn ^- isolates, notably those harboring prn1ΔSS, significantly increased since the early 2000s, and Prn ^- isolates were subsequently found nationwide. Multilocus variable-number tandem repeat analysis (MLVA) revealed that 24 (73%) of 33 Prn ^- isolates belong to MLVA-186, and 6 and 3 Prn ^- isolates belong to MLVA-194 and MLVA-226, respectively. The 3 MLVA types are phylogenetically closely related, suggesting that the 2 Prn ^- clinical strains (harboring prn1ΔSS and prn1::IS481) have clonally expanded in Japan. Growth competition assays in vitro also demonstrated that Prn ^- isolates have a higher growth potential than the Prn ⁺ back-mutants from which they were derived. Our observations suggested that human host factors (genetic factors and immune status) that select for Prn ^- strains have arisen and that Prn expression is not essential for fitness under these conditions.