Summary The avian pathogen Ornithobacterium rhinotracheale (ORT) has been implied in the etiology of poultry respiratory disease in recent years. In order to evaluate whether Whatman Â® FTA Â® cards can be used for hazard-free transport and storage of ORT samples for posterior DNA amplification, a controlled assay was performed. Three ten-fold dilutions of an ORT culture suspension were spotted on FTA Â® cards and stored at room temperature (RT) for a period of 6 months. Sterile swabs were immersed in the same three ten-fold culture dilutions and stored at RT, 4ÂºC and -20ÂºC without storage medium for the same time period. DNA was extracted from both the FTA Â® cards and swabs one day, one week, 6 weeks and 6 months following sample preparation and stored at -20ÂºC. At the end of the experiment, semi-quantitative polymerase chain reaction (qPCR) amplification of the 16S rRNA gene was performed from DNA extracted throughout a 6-month period from all ORT samples stored on both FTA Â® cards and swabs. The obtained threshold cycle (Ct) values for each ORT DNA extraction date were within the same range for all samples in a dilution-dependent fashion, regardless of storage temperature or used material. Pure ORT colonies could be reisolated one day after sample preparation from the swab dilutions stored at all temperatures but not from the FTA Â® cards. We conclude that the efficiency of ORT DNA amplification from samples stored on FTA Â® cards or in swabs is similar. However, FTA Â® cards have the advantage of preventing microrganism growth, thus allowing safe transport and storage for at least 6 months for bacterial dilutions down to at least 10 4 â 10 5 cfu/ml.
WhatmanÂ® FTAÂ® cards performance for Ornithobacterium rhinotracheale DNA amplification
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InÃªs Berenguer Veiga, Kristin MÃ¼hldorfer, Hafez Mohamed Hafez, DÃ¶rte LÃ¼schow; WhatmanÂ® FTAÂ® cards performance for Ornithobacterium rhinotracheale DNA amplification. Avian Dis doi: https://doi.org/10.1637/aviandiseases-D-20-00030
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