Resumo

Bordetella pertussis is a human pathogens which causes pertussis (whooping cough), a very contagious respiratory disease. WHO estimates that in 2008 about 16 million cases of pertussis occurred worldwide, killing 195,000 children. In 2011, global official numbers of reported cases was 162,047. Vaccination is the most effective intervention to prevent pertussis. Resurgence of pertussis has been recently observed in some countries with high vaccination coverage. The Brazilian National Immunization Program (BNIP) uses the whole cell pertussis vaccines produced at Instituto Butantan. Despite the reduction in incidence of whooping cough after the introduction of vaccination, the risk of outbreaks remains a public health concern. We reported here the draft genome sequence of Butantan´sB. pertussis strain used to compose combined vaccines such as DTwP (Diphteria, Tetanus and Pertussis) for the BNIP. The sequencing was performed combining shotgun sequencing in 454 GS JR and on MiSeq-illumina. A total of 297,811 reads comprising 128Mbases with 32-fold coverage of the genome was generated using two shotgun sequencing in 454 GS JR system. The MiSeq sequencing generated 270,011 reads (39Mbases). A total of 829,782 reads were assembled in 288 contigs, using CLC Genomics Workbench 3.6.5, the longest contig had an 84,859pb, the total clustered contig was 4,7Mbases and an average GC content 67.65%. When the reads were assembled with Tohama genome as reference, 1,170,111 reads were matched and 93,970 were not aligned. The draft genome of Butantan´s B. pertussis contains 3439 protein-coding sequences, 51 tRNA and 9 rRNA genes. Compared with the published genome of Tohama I strain, 30 genes were exclusively present in Butantan´s strain, one region contained 20 genes that could be found in B. pertussis CS, the Chinese vaccine strain, but not in Tohama strain. In contrast, 27 genes present in Tohama strain were absent in Butantan´s strain. These differences were mainly related with transcriptional regulatory and metabolic systems. Some small insertion re deletion events (indels) and 214SNPs were identified but need experimental confirmation. In conclusion, the genome sequence of a vaccinal B. pertussis strain used since the 80´s in the BNIP will provide the basis to understand the immune response against the whole cell pertussis vaccine as well as the molecular mechanisms in possible outbreaks due to the emergence of new pertussis strains.