В. Л. Колодкина, В. С. Мартынов
ГУ «Республиканский научно-практический центр эпидемиологии и микробиологии»
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
V. L. Kolodkina, V. S. Martynov
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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2. Centers for Disease Control and Prevention. 2015. Epidemiology and prevention of vaccine-preventable diseases, 13th ed. Public Health Foundation, Washington, DC.
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9. Pittet, L. F., Posfay-Barbe K. M. Bordetella holmesii: still emerging and elusive 20 years on // Emerging Infections. – 2016. – № 10. – С. 239–254.
10. van der Zee, A., Schellekens J. F. P., Mooi F. R. Laboratory diagnosis of pertussis. Clin Microbiol Rev. – 2015. – № 28. – Р. 1005–1026. – https://doi.org/10.1128/CMR .00031-15.
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12. Kerr, J. R., Matthews, R. C. Bordetella pertussis infection: pathogenesis, diagnosis, management, and the role of protective immunity // Eur. J. Clin. Microbiol. Infect. Dis. – 2000. – Vol. 19. –P. 77–88.
13. Reischl, U., Lehn N., Sanden G. N., Loeffelholz M. J.Real-Time PCR Assay Targeting IS481of Bordetella pertussis and Molecular Basis for Detecting Bordetella holmesii // Journal of Clinical Microbiology. – 2001. – Т. 39, № 5. – С. 1963–1966. – https://doi.org/10.1128/JCM.39.5.1963–1966.2001 PMID: 11326023.
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15. Kolodkina, V., Martynov V., Babenko A. Multiplex realtime PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis // Iran J. Microbiol. – 2014. – Vol. 6, № 3. – P. 140–148. 16. Diagnosis of whooping cough in Switzerland: diff erentiating Bordetella pertussis from Bordetella holmesii by polymerase chain reaction / L. F. Pittet [et al.] // PLoS One. – 2014. – Vol. 9, № З. – Р. e88936.
17. Antila, M. et al. Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis // Journal of medical microbiology. – 2006. – Т. 55, № 8. – С. 1043–1051.
18. Van den Bossche, D. et al. Prevalence of Bordetella holmesii and Bordetella bronchiseptica in respiratory tract samples from Belgian patients with pertussis-like symptoms by sensitive culture method and mass spectrometry // Acta Clinica Belgica. – 2013. – Т. 68, № 5. – С. 341–348.
19. Martini, H. et al. Improving specificity of Bordetella pertussis detection using a four target real-time PCR // PLoS One. – 2017. – Т. 12, № 4. – С. e0175587.
20. Mir-Cros, A. et al. Emergence of Bordetella holmesii as a causative agent of whooping cough, Barcelona, Spain // Emerging infectious diseases. – 2017. – Т. 23, № 11. – С. 1856.
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. – Т. 49, № 12. – С. 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. – Р. 322–331. – https://doi.org/10.1093/cid/cis888.
References
1. Mattoo, S., Cherry J. D. Molecular pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies // Clinical microbiology reviews. – 2005. – T. 18, № 2. – S. 326–382.
2. Centers for Disease Control and Prevention. 2015. Epidemiology and prevention of vaccine-preventable diseases, 13th ed. Public Health Foundation, Washington, DC.
3. Barlow, R. S., Reynolds L. E., Cieslak P. R., Sullivan A. D. Vaccinated children and adolescents with pertussis infections experience reduced illness severity and duration, Oregon, 2010–2012 // Clin. Infect. Dis. – 2014. – Vol. 58(11). – P. 1523–9. – https://doi.org/10.1093/cid/ciu156 PMID: 24633685
4. Bouchez, V., Guiso N. Bordetella pertussis, B. parapertussis, vaccines and cycles of whooping cough //Pathogens and disease. – 2015. – T. 73, № 7.
5. Mattoo, S., Foreman-Wykert A. K., Cotter P. A., Miller J. F. Mechanisms of Bordetella pathogenesis // Front. Biosci. – 2001. – T. 6, № 1.
6. Cherry, J. D., Seaton B. L. Patterns of Bordetella parapertussis respiratory illnesses: 2008–2010 // Clinical infectious diseases. – 2012. – T. 54, № 4. – S. 534–537. – https://doi.org/10.1093/cid/cir860 PMID: 22156857
7. Zhang, X., Weyrich L. S., Lavine J. S., Karanikas A. T., Harvill E. T. Lack of cross-protection against Bordetella holmesii after pertussis vaccination // Emerging infectious diseases. – 2012. – T. 18, № 11. – S. 1771. – https://doi.org/10.3201/ eid1811.111544 PMID: 23092514
8. Pittet, L, F., Emonet S., Franzois P., Bonetti E.-J., Schrenzel J., Hug M., Altwegg M., Siegrist C.-A., Posfay-Barbe K. M. Diagnosis of whooping cough in Switzerland: differentiating Bordetella pertussis from Bordetella holmesii by polymerase chain reaction // PLoS One. – 2014. – № 9. – P. e88936. – https://doi.org/10.1371/journal.pone.0088936.
9. Pittet, L. F., Posfay-Barbe K. M. Bordetella holmesii: still emerging and elusive 20 years on // Emerging Infections. – 2016. – № 10. – S. 239–254.
10. van der Zee, A., Schellekens J. F. P., Mooi F. R. Laboratory diagnosis of pertussis. Clin Microbiol Rev. – 2015. – № 28. – P. 1005–1026. – https://doi.org/10.1128/CMR.00031-15.
11. Muller, F. M, Hoppe J. E,Wirsing von Konig C. H. Laboratory diagnosis of pertussis: state of the art in 1997 // J Clin Microbiol. – 1997. – Vol. 35. – P. 2435–2443.
12. Kerr, J. R., Matthews R. C. Bordetella pertussis infection: pathogenesis, diagnosis, management, and the role of protective immunity // Eur J Clin Microbiol Infect Dis. – 2000. – Vol. 19. – P. 77–88.
13. Reischl, U., Lehn N., Sanden G. N., Loeffelholz M. J. Real-Time PCR Assay Targeting IS481of Bordetella pertussis and Molecular Basis for Detecting Bordetella holmesii // Journal of Clinical Microbiology. – 2001. – T. 39, № 5. – S. 1963–1966. – https://doi.org/10.1128/JCM.39.5.1963-1966.2001 PMID: 11326023.
14. Tatti, K. M., Sparks K. N., Boney K. O., Tondella M. L. Novel multitarget real-time PCR assay for rapid detection of Bordetella species in clinical specimens // Journal of clinical microbiology. – 2011. – T. 49, № 12. – S. 4059–4066. – https://doi.org/ 10.1128/JCM.00601-11 PMID: 21940464
15. Kolodkina, V., Martynov V., Babenko A. Multiplex realtime PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis // Iran J. Microbiol. – 2014. – Vol. 6, № 3. – P. 140–148.
16. Diagnosis of whooping cough in Switzerland: diff erentiating Bordetella pertussis from Bordetella holmesii by polymerase chain reaction / L. F. Pittet [et al.] // PLoS One. – 2014. – Vol. 9. – P. e88936.
17. Antila, M. et al. Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis // Journal of medical microbiology. – 2006. – T. 55, № 8. – S. 1043–1051.
18. Van den Bossche, D. et al. Prevalence of Bordetella holmesii and Bordetella bronchiseptica in respiratory tract samples from Belgian patients with pertussis-like symptoms by sensitive culture method and mass spectrometry //Acta Clinica Belgica. – 2013. – T. 68, № 5. – S. 341–348.
19. Martini, H. et al. Improving specificity of Bordetella pertussis detection using a four target real-time PCR // PLoS One. – 2017. – T. 12, № 4. – S. e0175587.215. Mir-Cros, A. et al. Emergence of Bordetella holmesii as a causative agent of whooping cough, Barcelona, Spain // Emerging infectious diseases. – 2017. – T. 23, № 11. – S. 1856.
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.
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