Cargando…

Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions

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 it...

Descripción completa

Detalles Bibliográficos
Autores principales: Shalaby, Azhar G., Bakry, Neveen R., Mohamed, Abeer A. E., Khalil, Ashraf A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Veterinary World 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7704315/
https://www.ncbi.nlm.nih.gov/pubmed/33281363
http://dx.doi.org/10.14202/vetworld.2020.2243-2251
_version_ 1783616796966780928
author Shalaby, Azhar G.
Bakry, Neveen R.
Mohamed, Abeer A. E.
Khalil, Ashraf A.
author_facet Shalaby, Azhar G.
Bakry, Neveen R.
Mohamed, Abeer A. E.
Khalil, Ashraf A.
author_sort Shalaby, Azhar G.
collection PubMed
description 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 10(4) to 10(7) (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.
format Online
Article
Text
id pubmed-7704315
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Veterinary World
record_format MEDLINE/PubMed
spelling pubmed-77043152020-12-05 Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions Shalaby, Azhar G. Bakry, Neveen R. Mohamed, Abeer A. E. Khalil, Ashraf A. Vet World Research Article 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 10(4) to 10(7) (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. Veterinary World 2020-10 2020-10-28 /pmc/articles/PMC7704315/ /pubmed/33281363 http://dx.doi.org/10.14202/vetworld.2020.2243-2251 Text en Copyright: © Shalaby, et al. http://creativecommons.org/licenses/by/4.0 Open Access. This article is 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.
spellingShingle Research Article
Shalaby, Azhar G.
Bakry, Neveen R.
Mohamed, Abeer A. E.
Khalil, Ashraf A.
Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title_full Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title_fullStr Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title_full_unstemmed Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title_short Evaluating Flinders Technology Associates card for transporting bacterial isolates and retrieval of bacterial DNA after various storage conditions
title_sort evaluating flinders technology associates card for transporting bacterial isolates and retrieval of bacterial dna after various storage conditions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7704315/
https://www.ncbi.nlm.nih.gov/pubmed/33281363
http://dx.doi.org/10.14202/vetworld.2020.2243-2251
work_keys_str_mv AT shalabyazharg evaluatingflinderstechnologyassociatescardfortransportingbacterialisolatesandretrievalofbacterialdnaaftervariousstorageconditions
AT bakryneveenr evaluatingflinderstechnologyassociatescardfortransportingbacterialisolatesandretrievalofbacterialdnaaftervariousstorageconditions
AT mohamedabeerae evaluatingflinderstechnologyassociatescardfortransportingbacterialisolatesandretrievalofbacterialdnaaftervariousstorageconditions
AT khalilashrafa evaluatingflinderstechnologyassociatescardfortransportingbacterialisolatesandretrievalofbacterialdnaaftervariousstorageconditions