Assessment of FTA card employment for Pasteurella multocida DNA transport and detection of virulence-associated genes in strains isolated from fowl cholera in the United States

Almeida, Cristiane Maria Galdino de; Furian, Thales Quedi; Borges, Karen Apellanis; Perdoncini, Gustavo; Mauel, Michael J.; Rocha, Sílvio Luis da Silveira; Nascimento, Vladimir Pinheiro do; Salle, Carlos Tadeu Pippi; Moraes, Hamilton Luiz de Souza

doi:10.1590/1678-4162-9821

Cited by 3 publications

(5 citation statements)

References 27 publications

(52 reference statements)

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“…As all bacterial DNA have been confirmed, and the results matched with that of Almeida et al [19], who detected P. multocida DNA in 100% of the samples and concluded that the FTA cards can be used for the storage period up to 35 days at different temperatures (4°C and 37°C).…”

Section: Discussionsupporting

confidence: 84%

“…The long-term DNA storage with high fidelity at various temperatures makes the FTA cards a good alternative way for gathering samples and suggested a way for the suitability of storing and transporting nucleic acid profitably for further molecular application [19]. Moreover, the demonstration of the FTA cards of overall agreement in detecting HPV level and genotyping when compared with other transport media also showed high-quality nucleic acid required for testing the molecular technique when use different methods to describe the nucleic acid from different samples and tissues by different methods for downstream molecular applications [48].…”

Section: Discussionmentioning

confidence: 99%

“…Escherichia coli is also one of the pathogenic agents in avian species [16] causing calf diarrhea [17], and another infectious agent is Pseudomonas aeruginosa [18]. Furthermore, FTA card commercial tools are possible tools for detecting Pasteurella multocida DNA in all samples tested and for storage period up to 35 days at 4°C and 37°C [19].…”

Section: Introductionmentioning

confidence: 99%

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

Shalaby

Bakry

Mohamed³

et al. 2020

Vet World

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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.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Shalaby

Bakry

Mohamed³

et al. 2020

Vet World

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“…These results indicate that both methods are feasible for viral RNA isolation on FTA cards. These findings are consistent with the results of Abdelwhab et al [8] and Perozo et al [19], who used TE buffer with a commercial extraction kit for AIV and NDV, as well as the results of Almeida et al [20], who used FTA purification reagent for Pasteurella multocida. However, to decide on the appropriate isolation method, some factors need to be considered, including the cost of the reagent, availability of the reagent, and the sensitivity of detection.…”

Section: B Asupporting

confidence: 91%

Sensitivity of RNA viral nucleic acid-based detection of avian influenza virus, Newcastle disease virus, and African horse sickness virus on flinders technology associates card using conventional reverse-transcription polymerase chain reaction

et al. 2022

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Background and Aim: The flinders technology associates (FTA) card is a cotton-based cellulose membrane impregnated with a chaotropic agent that inactivates infectious microorganisms, lyses cellular material, and fixes DNA and/or RNA within the fiber matrix. However, little is known about the effectiveness of these cards for detecting RNA viruses in animals. This study aimed to evaluate the sensitivity of RNA virus detection using conventional reverse-transcription polymerase chain reaction (RT-PCR) on FTA cards. Materials and Methods: A highly virulent Newcastle disease virus (NDV) and an avian influenza virus (AIV) with low pathogenicity were propagated using chicken embryonic eggs. Three days after inoculation, the allantoic fluid was harvested, stored at –80°C, and the stock virus was tested for virus titration. African horse sickness virus (AHSV) was obtained from a live attenuated vaccine that was dissolved and stored at –80°C. For sample preparation, each stock virus was 10-fold serially diluted and each dilution was inoculated onto an FTA card, followed by drying in a Class II safety cabinet. Both the stock virus and infected FTA card were genomically isolated using an extraction kit, FTA purification kit, and extraction kit with Tris-EDTA (TE) buffer. The target genome was then detected by one-step RT-PCR for NDV and AIV, and two-step RT-PCR for African horse sickness, including gel electrophoresis for the detection of specific nucleic acids. Results: The detection limit of stock AIV was compared on FTA cards, using the FTA purification kit, and with TE buffer with an extraction kit. The corresponding results were 1.47, 1.17, and 2.18 log10 EID50, respectively, while for NDV the results were 4.13, 4.83, and 4.84 log10 ELD50. Finally, detection limit of stock AHSV and AHSV on the FTA card extracted using TE buffer with an extraction kit were 4.30 and 4.01 log10 plaque-forming units, respectively. Conclusion: This study demonstrated that the detection limit or sensitivity of all tested RNA viruses on FTA cards did not differ when compared with those of the stock virus and in both methods for RNA isolation on FTA cards. These cards are suitable for collecting and transporting samples infected with RNA viruses, particularly AIV, NDV, and AHSV. Flinders technology associates cards also provide hazard-free samples, a reliable source of RNA for molecular characterization, and sufficient quantity for diagnostic applications based on nucleic acid-based detection.

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“…For example, FTA R cards have been used to transport samples of oral fluid, whole blood, sera, and nasal secretions containing the porcine reproductive and respiratory syndrome virus (PRRSV), as well as whole blood for swine influenza and African swine fever (17)(18)(19)(20)(21). Additionally, FTA R cards have been implemented in the diagnosis of numerous veterinary bacterial pathogens (21)(22)(23)(24)(25), including members of the Pasteurellaceae (16,26), but not (to our knowledge) A. pleuropneumoniae. Therefore, in order to reduce biosafety concerns and enable rapid and affordable diagnostics of this important swine pathogen, we have developed and validated an FTA R card protocol for processing clinical samples or bacterial cultures, which can be coupled to our previously described APP-mPCRs (10,12) for molecular detection and typing of A. pleuropneumoniae.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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Actinobacillus pleuropneumoniae (APP), the causative agent of porcine pleuropneumonia, is highly contagious and responsible for high morbidity, mortality, and economic losses in the swine industry worldwide, but quick serotyping and diagnosis are still not widely available. In this study, we sought to validate the use of Whatman FTA® cards for collection and processing of A. pleuropneumoniae isolates, or porcine lung tissue samples, for direct use in diagnostic multiplex PCRs. We have optimized the processing of 3-mm discs punched from FTA® cards loaded with cultured A. pleuropneumoniae, or imprinted on lesioned regions of lung tissue, with only three distilled water washes before addition into our APP-multiplex PCR (mPCR) assay for rapid, low-cost identification and serotyping. DNA captured on FTA® cards generated the same diagnostic PCR results as DNA extracted using commercial kits for 85 A. pleuropneumoniae clinical isolate cultures and 22 lung samples. Additionally, bacterial DNA bound to FTA® cards was detectable by PCR after 6 months of storage at 37°C. This study provides simple, efficient, rapid, and practical sample processing for detection and molecular serotyping of A. pleuropneumoniae.

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Assessment of FTA card employment for Pasteurella multocida DNA transport and detection of virulence-associated genes in strains isolated from fowl cholera in the United States

Cited by 3 publications

References 27 publications

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

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

Sensitivity of RNA viral nucleic acid-based detection of avian influenza virus, Newcastle disease virus, and African horse sickness virus on flinders technology associates card using conventional reverse-transcription polymerase chain reaction

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

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