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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Tehran University of Medical Sciences
2014-06-01
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doaj-a80a712c6a38476eaf7b6329c10679352020-12-02T05:59:18ZengTehran 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 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Valentina Kolodkina Vladimir Martinov Andrey Babenko |
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 |
author_facet |
Valentina Kolodkina Vladimir Martinov Andrey Babenko |
author_sort |
Valentina Kolodkina |
title |
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_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_sort |
multiplex real-time pcr assay for detection and differentiation of bordetella pertussis and bordetella parapertussis |
publisher |
Tehran University of Medical Sciences |
series |
Iranian Journal of Microbiology |
issn |
2008-3289 2008-4447 |
publishDate |
2014-06-01 |
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.
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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 |
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