Vet World Vol.13 October-2020 Article-29
Research Article
Veterinary World, 13(10): 2243-2251
https://doi.org/10.14202/vetworld.2020.2243-2251
Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
2. Department of Epidemiology Unit, Reference Laboratory for Veterinary Quality Control on Poultry Production, Animal Health Research Institute, Agricultural Research Centre, Dokki, Giza, Egypt.
3. Department of Buffalo Diseases, Animal Health Research Institute, Dokki, Giza, Egypt.
4. Institute of Biotechnology and Genetic Engineering, City of Scientific Research and Technology Applications, Borg Elarab, Alexandria, Egypt.
Background and Aim: Flinders Technology Associates (FTA) cards simplify sample storage, transport, and extraction by reducing cost and time for diagnosis. This study evaluated the FTA suitability for safe transport and storage of Gram-positive and Gram-negative bacterial cells of animal origin on its liquid culture form and from organ impression smears (tissues) under the same routine condition of microbiological laboratory along with detecting their nucleic acid over different storage conditions.
Materials and Methods: Increase in bacterial count from 104 to 107 (colony-forming units/mL) of 78 isolates representing seven bacterial species was applied onto cards. FTA cards were grouped and inoculated by these bacteria and then stored at different conditions of 24-27°C, 4°C, and –20°C for 24 h, for 2 weeks, for 1 and 3 month storage, respectively. Bacteriological examination was done, after which bacterial DNA was identified using specific primers for each bacterial type and detected by polymerase chain reaction (PCR).
Results: The total percentage of recovered bacteria from FTA cards was 66.7% at 24-27–C for 24 h, the detection limit was 100% in Gram-positive species, while it was 57.4% in Gram-negative ones. Regarding viable cell detection from organ impression smears, it was successful under the previous conditions. No live bacterial cells were observed by bacteriological isolation rather than only at 24-27°C for 24 h storage. All bacterial DNA were sufficiently confirmed by the PCR technique at different conditions.
Conclusion: Overall, the FTA card method was observed to be a valid tool for nucleic acid purification for bacteria of animal origin in the form of culture or organ smears regardless of its Gram type and is used for a short time only 24 h for storage and transport of live bacteria specifically Gram-positive type. Moreover, the bacterial nucleic acid was intact after storage in –20°C for 3 months and was PCR amplifiable. Keywords: bacteria, colony-forming units, Flinders Technology Associates, nucleic acid, polymerase chain reaction.
Keywords: bacteria, colony-forming units, Flinders Technology Associates, nucleic acid, polymerase chain reaction.
How to cite this article: Shalaby AG, Bakry NR, Mohamed AAE, Khalil AA (2020) Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions, Veterinary World, 13(10): 2243-2251.
Received: 03-07-2020 Accepted: 21-09-2020 Published online: 28-10-2020
Corresponding author: Azhar G. Shalaby E-mail: azhar_gaber@yahoo.com
DOI: 10.14202/vetworld.2020.2243-2251
Copyright: Shalaby, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.