Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis
Background and Objective: Rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among non-vaccinated infants for whom pertussis is life- threatening. Materials and Methods: Targets IS481, IS1001,...
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| Format: | Article |
| Language: | English |
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Tehran University of Medical Sciences
2014-06-01
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| Series: | Iranian Journal of Microbiology |
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| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/429 |
| _version_ | 1823987995668119552 |
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| author | Valentina Kolodkina Vladimir Martinov Andrey Babenko |
| author_facet | Valentina Kolodkina Vladimir Martinov Andrey Babenko |
| author_sort | Valentina Kolodkina |
| collection | DOAJ |
| description | Background and Objective: Rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among non-vaccinated infants for whom pertussis is life- threatening.
Materials and Methods: Targets IS481, IS1001, BP0026 and human GAPDH gene were used to develop a multiplex real- time PCR assay based on the TaqMan technology for detection and identification of Bordetella pertussis and Bordetella parapertussis in clinical samples. A total of 121 human clinical specimens obtained within 2012-2013 were used to evaluate the multiplex real-time PCR assay. Clinical specimens were also tested for culture and conventional PCR. Sensitivity and specificity for culture, conventional PCR, and multiplex real-time PCR were measured in comparison with a clinical standard for B. pertussis infection.
Results: The lower limit of detection (LLOD) of the multiplex assay was similar to the LLOD of each target in an individual assay format, which was approximately 1 genomic equivalent per reaction for IS481, IS1001 and 10 genomic equivalents per reaction for BP0026 target. When the B. pertussis assays were compared with a clinical standard for B. pertussis infection, sensitivity was 5, 59 and 89% the specificity was 100, 100 and 100% for culture, conventional PCR, and multiplex real-time PCR, respectively.
Conclusions: Developed multiplex real-time PCR offers a fast tool with high sensitivity and specificity for the diagnosis of B. pertussis and B. parapertussis infections which is suitable for implementation in a routine laboratory diagnostics. |
| first_indexed | 2024-12-18T13:36:44Z |
| format | Article |
| id | doaj.art-a80a712c6a38476eaf7b6329c1067935 |
| institution | Directory Open Access Journal |
| issn | 2008-3289 2008-4447 |
| language | English |
| last_indexed | 2024-12-18T13:36:44Z |
| publishDate | 2014-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Microbiology |
| spelling | doaj.art-a80a712c6a38476eaf7b6329c10679352022-12-21T21:06:00ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472014-06-0163Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussisValentina Kolodkina0Vladimir Martinov1Andrey Babenko2Republican Research & Practical Centre for Epidemiology and Microbiology, Minsk, Belarus.Republican Research & Practical Centre for Epidemiology and Microbiology, Minsk, Belarus.N. N. Aleksandrov Republican Scientific and Practical Centre of Oncology and Medical Radiology, Minsk, Belarus.Background and Objective: Rapid diagnosis of pertussis is important for the timely isolation of the infection source and early prevention measures among the contact persons, especially among non-vaccinated infants for whom pertussis is life- threatening. Materials and Methods: Targets IS481, IS1001, BP0026 and human GAPDH gene were used to develop a multiplex real- time PCR assay based on the TaqMan technology for detection and identification of Bordetella pertussis and Bordetella parapertussis in clinical samples. A total of 121 human clinical specimens obtained within 2012-2013 were used to evaluate the multiplex real-time PCR assay. Clinical specimens were also tested for culture and conventional PCR. Sensitivity and specificity for culture, conventional PCR, and multiplex real-time PCR were measured in comparison with a clinical standard for B. pertussis infection. Results: The lower limit of detection (LLOD) of the multiplex assay was similar to the LLOD of each target in an individual assay format, which was approximately 1 genomic equivalent per reaction for IS481, IS1001 and 10 genomic equivalents per reaction for BP0026 target. When the B. pertussis assays were compared with a clinical standard for B. pertussis infection, sensitivity was 5, 59 and 89% the specificity was 100, 100 and 100% for culture, conventional PCR, and multiplex real-time PCR, respectively. Conclusions: Developed multiplex real-time PCR offers a fast tool with high sensitivity and specificity for the diagnosis of B. pertussis and B. parapertussis infections which is suitable for implementation in a routine laboratory diagnostics.https://ijm.tums.ac.ir/index.php/ijm/article/view/429Bordetella parapertussisBordetella pertussisMultiplex real-time PCRdiagnosis |
| spellingShingle | Valentina Kolodkina Vladimir Martinov Andrey Babenko Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis Iranian Journal of Microbiology Bordetella parapertussis Bordetella pertussis Multiplex real-time PCR diagnosis |
| title | Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis |
| title_full | Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis |
| title_fullStr | Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis |
| title_full_unstemmed | Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis |
| title_short | Multiplex real-time PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis |
| title_sort | multiplex real time pcr assay for detection and differentiation of bordetella pertussis and bordetella parapertussis |
| topic | Bordetella parapertussis Bordetella pertussis Multiplex real-time PCR diagnosis |
| url | https://ijm.tums.ac.ir/index.php/ijm/article/view/429 |
| work_keys_str_mv | AT valentinakolodkina multiplexrealtimepcrassayfordetectionanddifferentiationofbordetellapertussisandbordetellaparapertussis AT vladimirmartinov multiplexrealtimepcrassayfordetectionanddifferentiationofbordetellapertussisandbordetellaparapertussis AT andreybabenko multiplexrealtimepcrassayfordetectionanddifferentiationofbordetellapertussisandbordetellaparapertussis |