fta cards
Recently Published Documents


TOTAL DOCUMENTS

93
(FIVE YEARS 43)

H-INDEX

12
(FIVE YEARS 4)

2022 ◽  
pp. 16-21
Author(s):  
Baraa Akeel Al-Hasan ◽  
Abdullah O. Alhatami ◽  
Husam Muhsen Abdulwahab ◽  
Ghadeer Sabah Bustani ◽  
Muhammad Ali Hameed ◽  
...  

Background and Aim: Swollen head syndrome (SHS) is a complex disease caused by various agents, including bacterial and viral pathogens, as well as environmental factors. Avian metapneumovirus (aMPV) is one of the most important causes of respiratory diseases and SHS in poultry and one of the most widespread viruses worldwide; however, it has not been recorded in Iraq. This study aimed at the molecular identification and subtyping of aMPV in poultry, with the objectives of investigating the prevalence of aMPV in infected broiler flocks with SHS and molecular typing using primers specific to the study of the prevalence of subtypes A, B, and C of aMPV. Materials and Methods: This study was performed on 67 broiler farms that reported typical SHS from September 2018 to August 2019. Swabs were collected from the trachea, infraorbital sinuses, and lung, then uploaded on FTA cards and subjected to an RNA extraction protocol. Results: aMPV was detected in 16 (23.8%) samples. Molecular typing using primers specific to the attachment glycoprotein (G) gene showed that all positive samples belonged to subtype B, as assessed using the real-time polymerase chain reaction technique. Conclusion: aMPV may be the main etiological factor causing SHS in poultry. Moreover, this was the first report of the prevalence of subtype B aMPV strains in broiler farms in Iraq.


2021 ◽  
Author(s):  
Grace Gysin ◽  
Plutarco Urbano ◽  
Luke Brandner-Garrod ◽  
Shahida Begum ◽  
Mojca Kristan ◽  
...  

Background: Accurate surveillance of triatomine household infestation is crucial for Chagas disease vector control. However, no gold standard detection method with high levels of sensitivity or specificity is currently available. Several intrinsic features of triatomine bug behaviour and the lifecycle of Trypanosoma (T.) cruzi lead to deposition of environmental DNA (eDNA) in infested houses. This study evaluated the use of FTA cards and cotton-tipped swabs as low-technology, cost-effective tools for simultaneous detection of T. cruzi and vector eDNA in the laboratory and field. Methods/Principal Findings: This study had two components: (1) laboratory evaluation and optimisation of QIAcard® FTA® classic cards to detect Rhodnius (R.) prolixus eDNA by altering five different environmental variables (darkness, triatomine number, temperature, feeding status and degradation at ambient temperature); (2) detection of R. prolixus and T. cruzi eDNA from cotton-tipped house wall swabs from an endemic region in Casanare Department, Colombia. eDNA was extracted from all specimens and amplified using a multiplex TaqMan qPCR assay targeting the R. prolixus 12S rRNA gene and T. cruzi satellite DNA region. R. prolixus eDNA from five 3rd/4th instar nymphs was successfully amplified from FTA cards after as little as 15 minutes of contact time under standard insectary conditions. Factors significantly increasing eDNA detection from FTA cards were increasing temperature from 21oC to 27-32oC, triatomine bug density from 1-25 bugs and recent blood-feeding. eDNA was detectable from FTA cards stored at room temperature for at least two weeks. In cotton-tipped swabs from the field, the sensitivity and specificity of R. prolixus eDNA detection was 60.6% (n=20/33) and 100% (n=33/33), respectively. T. cruzi eDNA was amplified from 93.9% (n=31/33) of infested houses. Conclusions/Significance: FTA cards are a highly sensitive tool for entomological surveillance of R. prolixus and exhibit little variability under different environmental conditions. Additionally, cotton-tipped swabs are a relatively sensitive tool for entomological and parasitological surveillance of R. prolixus and T. cruzi in situ, but more feasible due to low cost. Both methods could be utilised by citizen science initiatives to contribute to the control of Chagas disease in endemic communities.


Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3564
Author(s):  
Behailu Assefa Wayou ◽  
Gezahegne Mamo Kassa ◽  
Daniela Pasotto ◽  
Teshale Sori ◽  
Claudia Maria Tucciarone ◽  
...  

The importance of poultry production is globally increasing, in Ethiopia as well, where high-quality protein and contained costs make poultry a valuable food resource. However, this entails some problems linked to rural, backyard and intensively reared flock proximity and pathogen circulation. This study is aimed at monitoring the presence of important viral pathogens in poultry (infectious bronchitis virus (IBV), avian metapneumovirus (aMPV), infectious bursal disease virus (IBDV) and Newcastle disease virus (NDV)) in Ethiopia. Respiratory and cloacal swabs and bursa of Fabricius and kidney imprints on FTA cards were collected in 2021 from 16 farms and tested for IBV, aMPV, NDV and IBDV. One farm was positive for IBDV, resulting in strains similar to those present in vaccines, belonging to genogroup A1a; two farms were positive for IBV but, due to sensitivity limits, only one sample was sequenced, resulting in a 4/91-like strain (GI-13); a layer farm tested positive for NDV with a Lasota-like vaccine strain. These findings suggest a low presence of these pathogens, probably due to the implementation of vaccination strategies, which is also testified by the detection of vaccine strains. A close diagnostic activity should be implemented on a routine basis in order to monitor pathogen circulation, ameliorate biosecurity measures and protect animal health and production levels.


2021 ◽  
Vol 22 (23) ◽  
pp. 12915
Author(s):  
Ahmed Elnagar ◽  
Timm C. Harder ◽  
Sandra Blome ◽  
Martin Beer ◽  
Bernd Hoffmann

FTA cards and related products simplify the collection, transport, and transient storage of biological sample fluids. Here, we have compared the yield and quality of DNA and RNA released from seven different FTA cards using seven releasing/extraction methods with eleven experimental eluates. For the validation, dilution series of African swine fever virus (ASFV) positive EDTA blood and Influenza A virus (IAV) positive allantoic fluid were used. Based on our data, we conclude that direct PCR amplification without the need for additional nucleic acid extraction and purification could be suitable and more convenient for ASFV DNA release from FTA cards. In contrast, IAV RNA loads can be amplified from FTA card punches if a standard extraction procedure including a lysis step is applied. These differences between the amplifiable viral DNA and RNA after releasing and extraction are not influenced by the type of commercial FTA card or the eleven different nucleic acid releasing procedures used for the comparative analyses. In general, different commercial FTA cards were successfully used for the storage and recovery of the ASFV and IAV genetic material suitable for PCR. Nevertheless, the usage of optimized nucleic acid releasing protocols could improve the recovery of the viral genome of both viruses. Here, the application of Chelex® Resin 100 buffer mixed with 1 × Tris EDTA buffer (TE, pH 8.0) or with TED 10 (TE buffer and Dimethylsulfoxid) delivered the best results and can be used as a universal method for releasing viral DNA and RNA from FTA cards.


2021 ◽  
Vol 2 ◽  
Author(s):  
Philip A. Karlsson ◽  
Carolina Persson ◽  
James Akoko ◽  
Bernard Bett ◽  
Åke Lundkvist ◽  
...  

Brucella is a highly pathogenic bacteria endemic in Kenya, and in spite of its severity in humans, the highly inadequate Febrile Antigen Brucella Agglutination Test (FBAT) remains a primary tool for its diagnosis. Blood samples were collected from febrile patients in Kinna health center and screened by the local routine. Milk samples were purchased from local milk hawkers and analyzed for Brucella antibodies using the milk ring test (MRT). The MRT-positive milk was traced to farms, and lactating cattle were sampled for milk and blood. Milk was MRT-tested and the serum was analyzed using the Rose Bengal test (RBT) and iELISA. Available patient and farm samples were stored on FTA cards for qPCR analyses. Despite a limited sample size, our study, in line with previous reports, shows a low diagnostic sensitivity (67%) and specificity (40%) of FBAT when compared to qPCR. As many as 48% of the raw bulk cattle milk samples were MRT-positive for Brucella antibodies and 60% of cattle on three visited farms were IS711 qPCR-positive. This case-based One Health investigation confirms the suspected Brucella presence, suggesting a targeted vaccination at high-prevalence farms, urgent interventions on milk safety, and a re-evaluation of the diagnostic and treatment regimen.


Author(s):  
Michele D Tisdale ◽  
David R Tribble ◽  
Indrani Mitra ◽  
Kalyani Telu ◽  
Huai-Ching Kuo ◽  
...  

Abstract Background: We assessed the compliance with self-collection of stool smears on Whatman® FTA® Elute Card (FTA Card) and detection of travelers’ diarrhea (TD) associated pathogens using a quantitative PCR assay (customized TaqMan® array card [TAC]), in a prospective, observational cohort of travelers. Methods: Enrolled travelers documented symptoms on a travel diary and collected an FTA Card during a diarrheal episode, or at the end of travel if they remained asymptomatic. TAC testing was performed on FTA Cards from TD cases and 1:1 matched asymptomatic controls and 1:1 matched loose stool cases that did not meet TD criteria. Odds ratios (OR) were used to determine the association between detected pathogens and TD. Results: 484 of 2456 (19.7%) travelers completed an illness diary and met TD criteria, and 257 (53.1%) collected an FTA Card during the TD episode. FTA Cards were stored for a median of 2 years at room temperature (IQR: 1-4 years) before extraction and testing. The overall TAC detection rate in TD cases was 58.8% (95%CI: 52.5-64.8). Enterotoxigenic E. coli was the most common pathogen in TD cases (26.8%) and 3.5% of samples were positive for norovirus. The odds of detecting TD-associated pathogens in 231 matched cases and asymptomatic controls was 5.4 (95% CI: 3.6-8.1) and 2.0 (95% CI:1.1-3.7) in 121 matched TD and loose stool cases (p < 0.05). Enteroaggregative E coli was the most common pathogen detected in asymptomatic controls and loose stool cases. Detection of diarrheagenic E coli, Shigella/enteroinvasive E coli (EIEC), and Campylobacter spp. was significantly associated with TD. Conclusions: FTA Cards are a useful adjunct to traditional stool collection methods for evaluating the pathogen-specific epidemiology of TD in austere environments. Qualitative detection of pathogens was associated with TD. Measures to improve compliance and quality of FTA Card collection with decreased storage duration may further optimize detection.


2021 ◽  
pp. 2346-2355
Author(s):  
Baraa Akeel Al-Hasan ◽  
Abdullah O. Alhatami ◽  
Husam Muhsen Abdulwahab ◽  
Ghadeer Sabah Bustani ◽  
Eman Abdul Wahab Alkuwaity

Background and Aim: The swollen head syndrome (SHS) makes up complex diseases that infect the upper respiratory tract in poultry and causes several economic losses. Furthermore, this syndrome is considered one of the multifactorial etiological agents. Therefore, this study isolated and molecularly detected Ornithobacterium rhinotracheale (ORT) in poultry. Materials and Methods: This study was conducted at 67 broiler farms that had birds observed to be infected with the SHS from September 2018 until August 2019. Subsequently, swabs were collected from their trachea, infraorbital sinuses, and lungs, after which obtained samples were treated through two methods: (a) The direct method, by uploading samples on FTA cards, and the indirect method using a transport media. Afterward, reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the directly treated samples; howeverAQ1, the culture method, followed by PCR, was used to analyze the indirectly treated samples. Next, a partial 16S RNA gene was isolated using four positive PCR products, after which the effect of 16 antibiotics was studied on the seven local ORT strains isolated. Results: The quantity of ORT isolated using the direct method was 28 (41.7%) samples, which were all positive for the strain. Identification was by direct molecular identification (RT-PCR) from samples loaded on FTA cards. Alternatively, 7 (10.4%) ORTs were detected from the indirect method, as obtained using the culture method and biochemical tests. Then, PCR was subsequently used to confirm the results. As observed, 784 bp bands were shown for all seven ORT isolates. Furthermore, results revealed a significant difference in the detection of ORT strains between direct and indirect methods, with p-value (<0.05) and standard deviation of the error ±0.038 for the direct, then ±0.061 for the indirect method. For further analysis on the strain types, four 784 bp PCR products were taken, then partial 16S ribosomal sequence typing was conducted. All these four strains were found to be recorded in NCBI for the 1st time as a local Iraqi strain, with accession numbers (MN931657, MN931656, MN931655, and MN931654). Notably, results also showed that all isolated strains were multidrug-resistant. Conclusion: From the results, ORT is proposed to be implicated as one of the etiological factors that cause SHSs in poultry. Phylogenetic analysis of the current ORT bacterial strains also showed that they are closely related to the Egyptian isolates.


2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Noelle Fynmore ◽  
Renke Lühken ◽  
Heike Maisch ◽  
Tina Risch ◽  
Sabine Merz ◽  
...  

Abstract Background For over a decade, monitoring of West Nile virus (WNV) in Germany has consisted of a bird monitoring programme as well as a mosquito-based surveillance programme employing CO2-baited encephalitis vector surveillance (EVS) traps for mass trapping and screening of mosquitoes. In contrast to the EVS traps, the Reiter/Cummings type box gravid trap collects gravid female mosquitoes, which have already taken a blood meal, increasing the likelihood of being infected with pathogens. The traps can be equipped with a honey-baited Flinders Technology Associates® (FTA) card to encourage sugar feeding by the trapped mosquitoes. FTA cards contain nucleic acid preserving substances, which prevent the degradation of viral RNA in the expectorated mosquito saliva and allows for testing the card for flavivirus RNA. This study aimed to assess the suitability of the method for WNV surveillance in Germany as an alternative to previous methods, which are expensive, time-consuming, and predominantly target host-seeking populations less likely to be infected with WNV. Methods In the Thüringer Zoopark Erfurt, snowy owls (Nyctea scandiaca) and greater flamingos (Phoenicopterus roseus) died of WNV infections in July and August 2020. In response, five Reiter/Cummings type box gravid traps were positioned during the daytime on the 10th, 13th, and 16th of September in five different locations. The FTA cards and mosquitoes in the chamber were collected, kept in a cool chain, and further processed for virus detection using a modified generic flavivirus reverse transcription PCR. Results A total of 15 trappings during September collected a total of 259 female mosquitoes, 97% of which were Culex pipiens sensu lato, as well as 14 honey-baited FTA cards. Eight mosquitoes tested PCR-positive for WNV. Four FTA cards tested PCR-positive for mosquito-borne flaviviruses, two of which were confirmed as WNV, and the remaining two confirmed as Usutu virus. Conclusion The suitability of the FTA cards in preserving viral RNA in the field and rapid turnaround time from collection to result is combined with a simple, cost-effective, and highly specific trapping method to create an arbovirus surveillance system, which circumvents many of the difficulties of previous surveillance programmes that required the analysis of mosquitoes in the laboratory. Graphical Abstract


Author(s):  
Romain Mabon ◽  
Michèle Guibert ◽  
Roselyne Corbiere ◽  
Didier Andrivon

Mating type is a critical trait in heterothallic organisms. In plant pathogenic oomycetes, like the late blight pathogen Phytophthora infestans, it is usually identified through pairing between tester and candidate isolates, a method which is both laborious and applicable to live isolates only. Therefore, developing simple and fast PCR tests to reliably identify P. infestans mating types is of great interest for population genetic studies. A multiplex PCR assay combining the amplification of a locus diagnostic for P. infestans and of one diagnostic for the A1 mating type was developed and validated on a collection of 1441 samples, covering the current and past diversity of European P. infestans populations. These samples obtained from either freeze-dried mycelium or from FTA cards on which diseased leaflets had been pressed. The multiplex assay correctly identified mating types in 97.4 % of these samples. The main source of incorrect assignment was the lack of amplification of the A1 diagnostic allele, due to insufficient DNA quality and/or quantity in the reaction mix. This multiplex PCR, applicable to both live and stored material, thus constitutes a useful addition to the set of molecular tools available for population typing in P. infestans.


Author(s):  
Na Yue ◽  
Zichao Jia

The emergence of outbreaks of foodborne illness is closely associated with food contamination caused by various enteric pathogens, such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus. The control of enteric pathogens poses a challenge due to the fact that these pathogens can persist for a long period of time in the environment. The rapid detection of pathogenic organisms plays a crucial role in the prevention and identification of crises related to health, safety, and well-being. Improper sample handling and processing may influence the diagnostic efficacy and accuracy. The aim of the present study was to compare the preservation capacity for enteric bacteria between Whatman Flinders Technology Associates (FTA) cards and swabs for reverse transcription-quantitative PCR (RT-qPCR) detection. It was found that Whatman FTA cards exhibited an improved preservation capacity for five types (both laboratory and environmental strains) of enteric bacteria, including Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus for RT-qPCR detection. Hence, Whatman FTA cards may be a suitable tool for the routine isolation of foodborne bacteria for molecular diagnosis. Therefore, the use of Whatman FTA cards for sample collection and preservation may increase sensitivity and accuracy for bacteria isolation and diagnosis.


Sign in / Sign up

Export Citation Format

Share Document