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[Simultaneous screening method for Bordetella species by conventional PCR assay]
Yumi Akiyama1, Etsuko Saito1, Miki Enomoto1
1Public Health Science Research Center, Hyogo Prefectural Institute of Public Health and Consumer Sciences.
Summary
A new multiplex PCR method allows for the simultaneous detection of Bordetella pertussis, B. parapertussis, and B. holmesii. This cost-effective assay aids in the surveillance of whooping cough and related infections.
Area of Science:
- Microbiology
- Molecular Biology
- Public Health
Background:
- Pertussis, or whooping cough, is a highly contagious respiratory infection.
- Accurate and rapid identification of Bordetella species is crucial for effective disease surveillance and control.
- Existing diagnostic methods may lack the ability to differentiate between closely related Bordetella species simultaneously.
Purpose of the Study:
- To develop and validate a simultaneous screening method for detecting and differentiating Bordetella pertussis, Bordetella parapertussis, and Bordetella holmesii.
- To assess the utility of this multiplex assay in a real-world epidemiological surveillance setting.
Main Methods:
- Development of a multiplex PCR assay utilizing four pairs of primers targeting specific insertion sequences (IS481, IS1001, IS1002, and hIS1001).
- Determination of detection limits for each Bordetella species DNA (5 fg/µL).
- Application of the multiplex assay to clinical nasopharyngeal swab specimens from patients with pertussis-like symptoms.
Main Results:
- The multiplex PCR method demonstrated efficient detection and discrimination of the three Bordetella species.
- Low-level presence of B. pertussis and B. holmesii was confirmed using LAMP (Loop-mediated isothermal amplification).
- Out of 42 nasopharyngeal swabs, B. pertussis was detected in 12, with 8 cases linked to a nursery school outbreak.
Conclusions:
- The developed multiplex PCR assay provides a rapid, cost-effective, and simultaneous screening tool for Bordetella species.
- This method is valuable for epidemiological surveillance of infectious diseases, enabling efficient identification of B. pertussis, B. parapertussis, and B. holmesii.
- The assay facilitates the timely detection of outbreaks and informs public health interventions.
