Whatman® FTA® Cards Performance for Ornithobacterium rhinotracheale DNA Amplification
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Date of Publication
2020
Publication Type
Article
Division/Institute
Contributor
Mühldorfer, Kristin | |
Hafez, Hafez Mohamed | |
Lüschow, Dörte |
Series
Avian diseases
ISSN or ISBN (if monograph)
0005-2086
Publisher
American Association of Avian Pathologists
Language
English
Publisher DOI
PubMed ID
33570099
BORIS DOI
Description
The avian pathogen Ornithobacterium rhinotracheale (ORT) has been implied in the etiology of poultry respiratory disease in recent years. To evaluate whether Whatmant Flinders Technology Associates (FTAt) cards can be used for hazard-free transport and storage of ORT samples for posterior DNA amplification, a controlled assay was performed. Three 10-fold dilutions of an ORT culture suspension were spotted on FTA cards and stored at room temperature (RT) for 6 mo. Sterile swabs were immersed in the same three 10-fold culture dilutions and stored at RT and 4 and 20 C without storage medium for the same time. DNA was extracted from both the FTA cards and swabs 1 day, 1 and 6 wk, and 6 mo following sample preparation and stored at 20 C. At the end of the experiment, real-time PCR amplification of the 16S ribosomal RNA gene was performed from DNA extracted throughout a 6-mo period from all ORT samples stored on both FTA cards and swabs. The obtained threshold cycle 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 1 day after sample preparation from the swab dilutions stored at all temperatures but not from the FTA cards. We conclude that the efficiency of ORT DNA amplification from samples stored on FTA cards or in swabs is similar. However, FTA cards have the advantage of preventing microorganism growth, thus allowing safe transport and storage, for at least 6 mo, for bacterial dilutions down to at least 104 –105 colony-forming units/ml.