Literature DB >> 33570099

Whatman® FTA® Cards Performance for Ornithobacterium rhinotracheale DNA Amplification.

Inês Berenguer Veiga1, Kristin Mühldorfer2, Hafez Mohamed Hafez3, Dörte Lüschow3.   

Abstract

The avian pathogen Ornithobacterium rhinotracheale (ORT) has been implied in the etiology of poultry respiratory disease in recent years. To evaluate whether Whatman® Flinders Technology Associates (FTA®) 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.

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Keywords:  zzm321990 Ornithobacterium rhinotrachealezzm321990 ; Whatman® FTA® cards

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Year:  2020        PMID: 33570099     DOI: 10.1637/aviandiseases-D20-00030

Source DB:  PubMed          Journal:  Avian Dis        ISSN: 0005-2086            Impact factor:   1.577


  1 in total

1.  Optimizing Release of Nucleic Acids of African Swine Fever Virus and Influenza A Virus from FTA Cards.

Authors:  Ahmed Elnagar; Timm C Harder; Sandra Blome; Martin Beer; Bernd Hoffmann
Journal:  Int J Mol Sci       Date:  2021-11-29       Impact factor: 5.923

  1 in total

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