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

Abstract

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.

Rapid Detection and Typing of Actinobacillus pleuropneumoniae Serovars Directly From Clinical Samples: Combining FTA® Card Technology With Multiplex PCR

“…Diagnostic samples applied to FTA ® cards include cultured microorganisms and infected bodily fluids (e.g., blood, oral fluids, and mucosal secretions). Their use has been validated for a variety of viral and bacterial pathogens ( 24 , 31 , 32 ), and recently, Escherichia coli, Staphylococcus aureus , and Pasteurella multocida have been identified from impression smears of internal organs from chickens collected on FTA ® cards ( 33 ).…”

Evaluation of fecal sample collection methods for feline gut microbiome profiling: fecal loop vs. litter box

“…More than 10 previous studies have explored fecal collection and storage methods, examining variables such as temperature, storage duration at different temperatures, and the application of stabilizers like the OMNI-gene GUT kit, 95% ethanol, RNAlater, and other preservative solutions (Van der Waaij et al, 1994;Dominianni et al, 2014;Doukhanine et al, 2014;Flores et al, 2015;Loftfield et al, 2016;Song et al, 2016;Vogtmann et al, 2017;Wong et al, 2017;Burz et al, 2019;Conrads and Abdelbary, 2019;Papanicolas et al, 2019;Tap et al, 2019;Wu et al, 2019;Liang et al, 2020;Shalaby et al, 2020). While these studies have identified various methods to achieve stable microbial composition results, a universally accepted standard protocol has yet to emerge.…”

Detection of Enterotoxigenic Escherichia coli and Clostridia in the Aetiology of Neonatal Piglet Diarrhoea: Important Factors for Their Prevention

“…FTA (Flinders Technology Associates) cards provide important advantages, such as sample storage, transport, and extraction, leading to a decrease in the cost and time needed for molecular diagnosis. FTA cards have been widely used to extract and stabilize DNA from samples of human and swine clinical cases [ 41 , 42 , 43 , 44 , 45 , 46 ]. The method of FTA cards is reported to be a valid diagnostic tool, and it can be used for a short period (24 h) for the storage and transport of live bacteria, specifically Gram-positive types [ 44 ].…”

“…FTA cards have been widely used to extract and stabilize DNA from samples of human and swine clinical cases [ 41 , 42 , 43 , 44 , 45 , 46 ]. The method of FTA cards is reported to be a valid diagnostic tool, and it can be used for a short period (24 h) for the storage and transport of live bacteria, specifically Gram-positive types [ 44 ].…”