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Insertion sequences shared by Bordetella species and implications for the biological diagnosis of pertussis syndrome
A Tizolova1, N Guiso, S Guillot
1Department of Biochemistry and Microbiology, Institute of Chemical Technology, Technicka 5, 166 28 Prague 6, Czech Republic.
Abstract:
The molecular diagnosis of pertussis and parapertussis syndromes is based on the detection of insertion sequences (IS) 481 and 1001, respectively. However, these IS are also detected in the genomes of various Bordetella species, such that they are not specific for either B. pertussis or B. parapertussis. Therefore, we screened the genome of recently circulating isolates of Bordetella species to compare the prevalence of IS481, IS1001 and, also IS1002 with previously published data and to sequence all IS detected. We also investigated whether the numbers of IS481 and IS1001 copies vary in recently circulating isolates of the different Bordetella species. We used the polymerase chain reaction (PCR) method for screening the genome of circulating isolates and to prepare the fragments for sequencing. We used Southern blotting and quantitative real-time PCR for quantification of the numbers of IS. We found no significant diversity in the sequences of the IS harboured in the genomes of the Bordetella isolates screened, except for a 71-nucleotide deletion from IS1002 in B. bronchiseptica. The IS copy numbers in the genome of recently circulating isolates were similar to those in reference strains. Our results confirm that biological diagnosis targeting the IS481 and IS1001 elements are not specific and detect the species B. pertussis, B. holmesii and B. bronchiseptica (IS481), and B. parapertussis and B. bronchiseptica (IS1001).
Insights
Molecular diagnosis of pertussis and parapertussis using insertion sequences (IS) 481 and 1001 is not specific. These IS elements are found in multiple Bordetella species, impacting diagnostic accuracy.
Area of Science:
- Microbiology
- Genomics
- Molecular Diagnostics
Background:
- Molecular diagnosis of pertussis and parapertussis relies on detecting insertion sequences (IS) 481 and 1001.
- These insertion sequences are not exclusive to Bordetella pertussis or Bordetella parapertussis, being present in other Bordetella species.
Purpose of the Study:
- To assess the prevalence and sequence diversity of IS481, IS1001, and IS1002 in recently circulating Bordetella species.
- To investigate copy number variations of IS481 and IS1001 in these isolates.
- To determine the diagnostic specificity of IS481 and IS1001 for Bordetella species identification.
Main Methods:
- Polymerase chain reaction (PCR) for screening and fragment preparation for sequencing.
- DNA sequencing to analyze insertion sequence diversity.
- Southern blotting and quantitative real-time PCR for insertion sequence copy number quantification.
Main Results:
- No significant sequence diversity was observed in the insertion sequences across Bordetella isolates, with a minor exception in IS1002.
- Insertion sequence copy numbers in recent isolates were comparable to reference strains.
- IS481 was detected in B. pertussis, B. holmesii, and B. bronchiseptica; IS1001 was detected in B. parapertussis and B. bronchiseptica.
Conclusions:
- The molecular detection of IS481 and IS1001 is not specific for B. pertussis and B. parapertussis, respectively.
- These insertion sequences are present in other Bordetella species, including B. holmesii and B. bronchiseptica.
- Current diagnostic methods targeting these IS elements require careful interpretation due to cross-reactivity.
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