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  • Алгоритм видовой дифференциации возбудителей рода Bordetella и результаты его использования в Республике Беларусь

Алгоритм видовой дифференциации возбудителей рода Bordetella и результаты его использования в Республике Беларусь

В. Л. Колодкина, В. С. Мартынов ГУ «Республиканский научно-практический центр эпидемиологии и микробиологии»
2021 №4 | Новые технологии
Аннотация
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Проведена видовая дифференциация возбудителей рода Bordetella в носоглоточных мазках пациентов с подозрением на коклюш с использованием разработанных нами ПЦР в реальном времени: дуплексной TaqMan ПЦР, направленной на выявление B. pertussis и B. parapertussis, с праймерами к мишеням IS481, IS1001, BP0026 и GAPDH гена человека, и двух моноплексных TaqMan ПЦР, направленных на выявление B. holmesii и B. bronchiseptica c праймерами к мишеням hIS1001 и flaA. При исследовании 3130 носоглоточных мазков B. pertussis, B. Parapertussis и B. holmesii были выявлены в 29,5 %, 0,9 % и 0,4 % образцов соответственно. Разработанный алгоритм на основе трех TaqMan ПЦР оказался полезным инструментом в молекулярной диагностике коклюша. Его использование позволило установить коциркуляцию разных видов рода Bordetella, доминирование в циркуляции B. pertussis, низкий уровень циркуляции B. parapertussis, B. holmesii и отсутствие в циркуляции B. bronchiseptica в Республике Беларусь.
Ключевые слова:
Bordetella pertussis, B. parapertussis, B. holmesii, B. bronhiseptica, ПЦР реального времени, коклюш
The algorithm for species differentiation of Bordetella genus pathogens and the results of its use in the Republic of Belarus

Species-specific differentiation of Bordetella pathogens in nasopharyngeal swabs of patients with suspected whooping cough was carried out using developed by us real-time PCR: duplex TaqMan PCR aimed for detecting B. pertussis and B. parapertussis, with primers targeting IS481, IS1001, BP0026 and GAPDH as the human internal control and two singleplex TaqMan PCR aimed for detecting B. holmesii and B. bronchiseptica with primers targeting hIS1001 and flaA specific genes. Among the 3130 throat swab analyzed, B. pertussis, B. parapertussis and B. holmesii were detected in 29.5 %, 0.9 % and 0.4 % of samples respectively. The developed algorithm proved to be a useful molecular-genetic diagnostic tool for whooping cough, established cocirculation of different species of Bordetella genus and demonstrated the predominance of B. pertussis, low rate of B. parapertussis and B. holmesii and the absence of B. bronchiseptica in the circulation in the Republic of Belarus.

Keywords
Bordetella pertussis, B. parapertussis, B. holmesii, B. bronhiseptica, real-time PCR, whooping cough
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  • 20. Njamkepo, E. et al. Significant findin g o f Bordetella holmesii DNA in nasopharyngeal samples from French patients with suspected pertussis // Journal of clinical microbiology. – 2011. – T. 49, № 12. – S. 4347–4348.
  • 21. Rodgers, L., Martin S. W., Cohn A., Budd J., Marcon M., Terranella A., Mandal S., Salamon D., Leber A., Tondella M. L., Tatti K., Spicer K., Emanuel A., Koch E., McGlone L., Pawloski L., Lemaile-Williams M., Tucker N., Iyer R., Clark T. A., Diorio M. Epidemiologic and laboratory features of a large outbreak of pertussis-like illnesses associated with cocirculating Bordetella holmesii and Bordetella pertussis–Ohio, 2010–2011 // Clin Infect Dis. – 2013. – Vol. 56. – P. 322–331. – https://doi.org/10.1093/cid/cis888. Поступила 12.03.2021 г.