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, 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.