کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
4357050 | 1300026 | 2013 | 7 صفحه PDF | دانلود رایگان |
Detection of the lethal amphibian fungus Batrachochytrium dendrobatidis relies on PCR-based techniques. Although highly accurate and sensitive, these methods fail to distinguish between viable and dead cells. In this study a novel approach combining the DNA intercalating dye ethidium monoazide (EMA) and real-time PCR is presented that allows quantification of viable B. dendrobatidis cells without the need for culturing. The developed method is able to suppress real-time PCR signals of heat-killed B. dendrobatidis zoospores by 99.9 % and is able to discriminate viable from heat-killed B. dendrobatidis zoospores in mixed samples. Furthermore, the novel approach was applied to assess the antifungal activity of the veterinary antiseptic F10® Antiseptic Solution. This disinfectant killed B. dendrobatidis zoospores effectively within 1 min at concentrations as low as 1:6400.
► We describe a novel approach to quantify viable Batrachochytrium dendrobatidis cells.
► Fast and culture-independent viability testing of B. dendrobatidis is made possible.
► The method demonstrates the antifungal activity of F10® Antiseptic Solution.
Journal: Fungal Biology - Volume 117, Issue 2, February 2013, Pages 156–162