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0000025607 00000 n Our data confirms this finding as storing cards for three days at high humidity affected RNA preservation more than three days of high temperature. 0000050268 00000 n Additionally, IgM generated in response to a wild-type (WT) YFV infection is indistinguishable from IgM generated after vaccination, including in rare cases of vaccine-associated viscerotropic disease, making serological diagnosis during mass vaccination campaigns difficult [6]. FTA (Flinders Technology Associates) card (GE Whatman, Maidstone, Kent, United Kingdom) is a specialized filter paper which is impregnated with reagents promoting Yang Y., Garver L.S., Bingham K.M., Hang J., Jochim R.C., Davidson S.A., Richardson J.H., Jarman R.G. 0000004878 00000 n Writing review & editing, Affiliation Evaluation of nucleic acid stabilization products for ambient temperature shipping and storage of viral RNA and antibody in a dried whole blood format. 0000045774 00000 n 0000010337 00000 n We acknowledge Augusto Zuluaga Ph.D. (c) at Universidad Tecnolgica de Pereira for his help in data analysis and Dr. Matthew H. Collins at Hope Clinic of the Emory Vaccine Center, Division of Infectious Diseases, Department of Medicine, School of Medicine, Emory University for English revision. The potentially eligible articles were entered in a database, screened based on title and abstract and reviewed independently by two research team members. Analysis of an RNA-seq strand-specific library from an east timorese cucumber sample reveals a complete cucurbit aphid-borne yellows virus genome. et al. Yes identification of animals, insects, and transgenics. Dong L., Lin C., Li L., Wang M., Cui J., Feng R., Liu B., Wu Z., Lian J., Liao G., Chen W., Qiao Y. Simply apply your sample to the FTA Card. Resources, Yes https://doi.org/10.1371/journal.pntd.0010487.t003. Articles were screened by title and abstract, and after examination of inclusion and exclusion criteria, relevant information was extracted. WebProduct Details. ). As the molecular diagnosis of YFV becomes more routine in YF-endemic regions, RNA stabilization provided by FTA cards may become integral to ensuring accurate diagnosis. We found 175 records using the search strategy, and 11 additional records by checking references of the included articles. When the source material likely contains PCR inhibitors, an additional step of cleaning with 200L of Ethanol 70% can be performed (Roy and Nassuth, 2005; Inoue et al., 2007; Narayanan et al., 2010; Chang et al., 2011). A total of 95.7% of the included studies extracted total RNA before targeted amplification of the viral RNA, while the 4.3% directly isolated viral RNA from the FTA card. . It was performed in 55.1% of the assessed studies. . https://doi.org/10.1371/journal.pntd.0011027 and transmitted securely. Writing review & editing, Roles hb``a``[ ,'(70.x`djPx%ndmjD. 0000009134 00000 n Use of FTA filter paper for the molecular detection of Newcastle disease virus. 0000052098 00000 n 6. There was a significant difference in time of detection at 1 pfu/punch between both one and two (one day vs. two days 95% CI: 0.2, 3.0) and two and three days (two days vs. three days 95% CI: 2.8, 5.1) of preincubation in high humidity. Supervision, Experiments involving 17D-204 virus were performed according to biosafety level 2 requirements and experiments involving Asibi virus were performed according to biosafety level 3 safety requirements. This method is attractive for several applications, including research of bacterial genetics, as DNA can be maintained at ambient temperature (Reeve et al., 2018). The elution and subsequent purification of the RNA are the most widely used, and different elution solutions in a wide range of volumes, time, agitation and temperatures have been tested. WebApplication. Other types of DBS paper are available (e.g. Storage time and temperature and card manipulation and processing during viral isolation are other possible factors that could affect viral infectiousness. Note that this method did not include an RNA purification step (Roy and Nassuth, 2005; Inoue et al., 2007; Narayanan et al., 2010; Chang et al., 2011). 0000095887 00000 n This method was effective for amplification of fragments up to 1700bp. In this study, the YFall qRT-PCR assay [9] was used to show that WT and vaccine strains of YFV were equally detectable after inoculation onto FTA cards (S1 Fig). FTA cards were placed in an incubator at 37C for three days at 7580% humidity, then were transferred into plastic bags containing one or two silica 1-gram gel desiccation packets (Dry & dry, California, USA) and sealed. We assessed studies that used FTA cards for storage and later isolation of RNA of viral origin with subsequent amplification. As such, laboratory diagnosis is required to confirm YF. In this study, we investigated Whatman FTA cards as an alternative stabilization method of YFV RNA for use in molecular diagnosis. The first part sought to determine whether exposure of cards to high humidity post inoculation affected their ability to stabilize YF RNA. The initial experiments were conducted in a controlled climate (4055% humidity). Investigation, Using contrived specimens, linear regression analysis showed that RNA detection from a single 6mm FTA card punch was significantly less sensitive than traditional RNA extraction; however, pooling RNA extracted Improving vector-borne pathogen surveillance: a laboratory-based study exploring the potential to detect dengue virus and malaria parasites in mosquito saliva. Maina S., Edwards O.R., de Almeida L., Ximenes A., Jones R.A. WebData file 28-9843-54 AA FTA* cards FTA technology Whether youre in a lab or deep in a rain forest, Whatman* FTA provides a remarkably easy way to collect and isolate nucleic acid samples for analysis. WebFTA protocol FTA extractions on all 249 FTA cards were processed strictly following the manufacturers protocol. 0000030918 00000 n Writing original draft, More data is required to explain why the bursal disease virus remained infectious when such a wide variety of viruses were inactivated after spotting on FTA cards (Perozo et al., 2006; Narayanan et al., 2010; Abdelwhab et al., 2011; Awad et al., 2014; Jozwiak et al., 2016; Madhanmohan et al., 2016). Hall-Mendelin S., Hewitson G.R., Genge D., Burtonclay P.J., De Jong A.J., Pyke A.T., van den Hurk A.F. Federal government websites often end in .gov or .mil. 0000014426 00000 n This molecular assay is commonly used in the field and has been shown to be both sensitive and specific for WT and vaccine strains of YFV [8,9]. Individual punches were then placed into 1.5 mL tubes and stored as described until RNA extraction was performed using the QIAmp viral RNA extraction kit (Qiagen, California, USA). Non-indicating QIAcard FTA formats are used for the collection, transport and archiving of biological samples such as blood, cells and tissue. 0 The faecal material was immediately frozen on collection at 20 C prior to use in the subsequent experiments. %%EOF QIAcardQ FTA Gene (100) has been used for: molecular biology and amplifying the whole genome. Since the stability and integrity of viral RNA for molecular detection is a significant concern during fieldwork, this systematic review focused on the evaluation of FTA cards as a strategy for detection of these pathogens. National Library of Medicine Inoue R., Tsukahara T., Sunaba C., Itoh M., Ushida K. Simple and rapid detection of the porcine reproductive and respiratory syndrome virus from pig whole blood using filter paper. Keeler S.P., Ferro P.J., Brown J.D., Fang X., El-Attrache J., Poulson R., Jackwood M.W., Stallknecht D.E. 4. In many cases, the sensitivity of molecular assays was shown to decrease with the use of FTA cards [23,28,31]. ObjectiveThis systematic review aims to summarize the reported effectiveness of FTA cards for storage and transport of viral RNA, as well as the conditions for their handling and use in downstream processes. 0000011077 00000 n This cold-chain is required to prevent the degradation of YFV RNA but is often not realistic in YF-endemic regions where transmission season is accompanied by hot and humid weather [3]. Flinders Technology Associates (FTA) cards have been widely used for DNA preservation and analysis, particularly in forensic sciences and genetic studies (Hsiao et al., 1999; Smith and Burgoyne, 2004). The timing of punching, pre-inoculation or post-inoculation was tested to ensure that the pre-punch protocol did not impact RNA yield. Whatman FTA cards have been employed as a solution for other RNA viruses as they rapidly inactivate virus and stabilize genetic material [11,24,3032]. FOIA Complete genome sequences of the potyvirus sweet potato virus 2 from East Timor and Australia. Maina S., Edwards O.R., de Almeida L., Ximenes A., Jones R.A. The experiment was repeated with a different brand of desiccation packet (Whatman FTA, 1-gram packets) to compare the efficacy of 1-gram desiccation packets. When there was disagreement in the inclusion of a study, a third researcher made the final decision. The virus is responsible for periodic outbreaks of yellow fever (YF) disease, a hemorrhagic disease with a case fatality rate of 3060% in severe cases [1]. We have previously designed an qRT-PCR assay that distinguishes between the two strains using locked nucleic acid (LNA) technology and believe that the use of FTA cards to transport clinical samples during an outbreak could increase the successful use of both molecular assays. Methodology, FTA cards are designed for room temperature collection, shipment, archiving, and purification of nucleic acids from a wide variety of biological samples for PCR analysis. Accessibility The references of the included articles were also reviewed looking for eligible articles. In this study, every paper-based matrix allowed the isolation of viral RNA and infectious virus. PCR conditions: enough information concerning the methods and reagents used for PCR, including primers and cycling conditions. These cards are compact, easy to use and were shown to inactivate YFV and stabilize YFV RNA at high temperature and humidity. Moscoso H., Raybon E.O., Thayer S.G., Hofacre C.L. Experiments of viral RNA detection from FTA cards were considered successful if the RNA fragment of interest was finally amplified from cDNA (Retro-transcription, RT) and PCR irrespective of the fragment size or if the RNA isolation allowed further genome sequencing through NGS. With FTA Elute, In this study we optimize the use of Whatman FTA cards in conjunction with YFV molecular diagnostics and show that the incorporation of this widely available commercial product could lessen the requirement for cold-chain, which is often unavailable or incomplete. Direct viral RNA purification, and 3. Additionally, though we conducted an extensive search in different databases, and the screened works were searched through different institutional subscriptions, four papers were not accessible for full-text assessment. The resulting articles were initially selected by title and abstract to identify possibly eligible studies according to the selected criteria. 0000011191 00000 n Maina S., Edwards O.R., Barbetti M.J., de Almeida L., Ximenes A., Jones R.A. Dilutions were performed in PBS. Finally, the effect of desiccating cards that were previously exposed to humid conditions was assayed to establish if dry FTA cards were required to stabilize RNA after inoculation. Use of FTA cards to work with biospecimens appears to be relatively safe since isolation of infectious virus was rarely reported. The dotted line at Ct value 37 indicates the cut-off for YFV RNA positivity and is based on cut-offs used in the molecular diagnosis of YFV in the field. RNA was then extracted from punches made from the whole FTA card or the pre-punched samples. 0000049333 00000 n Web4 sample areas for application of up to 500 l sample volume per card. To test if this reduction of input volume significantly impacts the sensitivity of the assay, the limit of detection of 10 L virus applied to FTA punches was assayed and compared to a direct extraction from 10 L of virus and the gold standard of a direct extraction of 140 L of virus. Different methods for extraction and purification were represented in the literature. 0000030766 00000 n 1 During a YFV outbreak, molecular diagnostics are highly important as IgM generated in response to WT YFV infection and vaccination are indistinguishable via serology. Confirmation of viral inactivation was performed using institutionally approved protocols. Two brands of desiccant packets were tested and after two days of desiccation, there was no significant difference in the length of detection for either titer of YFV tested (S4 Table). Paper-based archiving of biological samples from fish for detecting betanodavirus. https://doi.org/10.1371/journal.pntd.0010487, Editor: Pedro F. C. Vasconcelos, Universidade do Estado do Para: Universidade do Estado do Para, BRAZIL, Received: January 15, 2022; Accepted: May 10, 2022; Published: June 15, 2022. Citation: Davis EH, Velez JO, Russell BJ, Basile AJ, Brault AC, Hughes HR (2022) Evaluation of Whatman FTA cards for the preservation of yellow fever virus RNA for use in molecular diagnostics. https://doi.org/10.1371/journal.pntd.0010487.t002. Linhares D.C., Rovira A., Torremorell M. Evaluation of flinders Technology associates cards for collection and transport of samples for detection of Porcine reproductive and respiratory syndrome virus by reverse transcription polymerase chain reaction. Viremia during YFV infection is variable and may be quite low depending on disease severity and the timing of sample collection. Bags were returned to the incubator for one, two or seven days prior to inoculation when FTA cards were punched and inoculated with 10 pfu/punch or 1 pfu/punch of 17D-204 virus. Due to the threat YF poses to public health, a single case of constitutes an outbreak, making accurate diagnosis paramount. YFV 17D-204 virus was inoculated onto FTA cards (130 L sample over entire card) or onto FTA card punches (10 L/ punch) and allowed to dry completely. The stability of RNA genomic material of many different viruses under a wide range of conditions further supports the advantage of FTA cards use, even for metagenomic analysis (Yang et al., 2015; Maina et al., 2017b). If the work tried complete genome sequencing the answer to this question was also scored with two. Converting mosquito surveillance to arbovirus surveillance with honey-baited nucleic acid preservation cards. RNA was extracted from the cards and assayed using YFall qRT-PCR primers. Grund E., Darissa O., Adam G. Application of FTA (R) cards to sample microbial plant pathogens for PCR and RT-PCR. View correction. The viruses were inoculated at the same titer (6log10 pfu/mL). Although this volume can be applied to the entire FTA card, RNA extraction protocols utilizing FTA cards require the card to be punched prior to RNA extraction, limiting the volume of sample that is extracted. Web5.1.2 Whatman Protein Saver Card #903 (Whatman #10534612; Fisher Scientific #NC9307519) Note: These guidelines refer specifically to DBS created from Whatman 903 paper. Narayanan M.S., Parthiban M., Sathiya P., Kumanan K. Molecular detection of Newcastle disease virus using flinders tehnology associates-PCR. It was shown that extracting RNA from two punches significantly increased the number of days RNA could be detected (Table 1). These studies attempted viral isolation from FTA cards impregnated with infected material using cell culture (Moscoso et al., 2006, Muthukrishnan et al., 2008, Keeler et al., 2012, Linhares et al., 2012, Bankamp et al., 2013, Awad et al., 2014, Tam et al., 2015, Madhanmohan et al., 2016), bioassays infecting specific pathogen-free embryonated chicken eggs (Moscoso et al., 2005, Perozo et al., 2006, Abdelwhab et al., 2011, Jozwiak et al., 2016) or both (Narayanan et al., 2010). FTA cards were incubated at 37C and high humidity (8085%) for one, two, and three days prior to inoculation cards were removed from the incubator, immediately punched, and inoculated with 10 pfu/mL or 1 pfu/mL of 17204 virus. The use of FTA cards could make the difference between complete loss of detectable RNA and successful detection in many instances. Although these assays are successfully used to accurately detect YF, there are important caveats that must be addressed. Maw M.T., Yamaguchi T., Kasanga C.J., Terasaki K., Fukushi H. A practical tissue sampling method using ordinary paper for molecular detection of infectious bursal disease virus RNA by RT-PCR. However, successful applications of these cards have been also reported for DNA viruses and bacteria (Dong et al., 2017; Reeve et al., 2018). 30 Dec 2022: Davis EH, Velez JO, Russell BJ, Basile AJ, Brault AC, et al. Nevertheless, YFV inoculated onto pre-desiccated cards could be detected days longer than cards that had been stored in humid conditions. 0000002070 00000 n The current WHO diagnostic algorithm calls for the molecular diagnosis of YF from serum samples, which are collected at clinics and diagnosis is completed at regional reference labs [25]. Finally, the biosafety measures required to protect researchers and clinical lab workers are considered. At room temperature, 100,000 pfu/punch (R2 = 0.68), 10,000 pfu/punch (R2 = 0.80), 1,000 pfu/punch (R2 = 0.64) and 100 pfu/punch (R2 = 0.82) could be detected for over two weeks (Fig 3A). BackgroundDetection and characterization of viral RNA pathogens from fieldwork are challenging due to the instability of the RNA molecule. Samples from animals accounted for 38.3% of the publications, which identified viruses that cause disease in poultry, wild birds, suids, or bovids. Cards incubated at high humidity showed no difference in the time limit for detection of positive Ct values when compared to dry cards incubated at low humidity (S2 Table). 0000004789 00000 n %PDF-1.5 % Then, FTA cards should be managed and stored following the required biosafety precautions. When pooling RNA from two punches, individual punches were placed into 70 L of PBS and vortexed. Desiccating FTA cards for longer was shown to be less effective, which is likely due to desiccation packets becoming fully saturated. Infection with other flaviviruses such as dengue (14), Zika, and West Nile viruses must be ruled out by additional serological testing and confirmed at regional reference laboratories. The use of FTA(R) filter papers for diagnosis of avian influenza virus. Points represent the average of two experiments where extracts from one punch were made and were fit using a linear regression. Jozwiak M., Wyrostek K., Domanska-Blicharz K., Olszewska-Tomczyk M., Smietanka K., Minta Z. application of FTA (R) cards for detection and storage of avian influenza virus. A total of 47 articles were included. Dilutions to make the standard curve were performed in PBS. For studies using pooled RNA from two punches, only 10 pfu/punch and 1 pfu/punch were utilized. 0000003150 00000 n One study showed that incomplete drying of FTA cards caused the most significant loss of RNA yield [23]. To determine if vaccine and WT strains of YFV could be detected after inoculation onto FTA cards, the vaccine substrain 17D-204, WT strain Asibi, and PBS as a negative control were inoculated onto FTA cards and assayed for YFV RNA. In experiments addressing the ability of FTA card methodology to stabilize YFV RNA at variable temperature, RNA could be detected for more than two weeks following storage at 25C. Use of FTA Technology to extract wheat streak mosaic virus and candidatus liberibacter solanacearum from single vectors. https://doi.org/10.1371/journal.pntd.0010487.g001. Hall-Mendelin S., Ritchie S.A., Johansen C.A., Zborowski P., Cortis G., Dandridge S., Hall R.A., Van den Hurk A.F. Awad F., Baylis M., Jones R.C., Ganapathy K. Evaluation of flinders technology associates cards for storage and molecular detection of avian metapneumoviruses. Limit of detection (LOD) and time to negative result were calculated by linear regression modeling, estimating when the mean Ct value reached the cut-off threshold of 37. WebProduct Details QIAcard FTA Indicating cards and QIAcard FTA Elute Indicating cards are designed to stabilize and protect nucleic acids at room temperature. Funding: The authors received no specific funding for this work. Funding acquisition, To prevent cross-contamination, the Harris Uni-Core tool was cleaned between each sample by punching on a fresh, unused FTA card. Smith L.M., Burgoyne L.A. It is possible that publication bias, the lack of publishing reports with negative results, could lead to failure in identifying studies that were unable to amplify RNA extracted from the FTA cards. Viremia can last the duration of infection in severe cases of YF but is highly variable [37,38]. For stability experiments, 17D-204 was diluted in PBS or human, flavivirus-negative serum and inoculated onto punches at concentrations of 7 log10 pfu/mL (100,000 pfu/punch), 6 log10 pfu/mL (10,000 pfu/punch), 5 log10 pfu/mL (1,000 pfu/punch), 4 log10 pfu/mL (100 pfu/punch), 3 log10 pfu/mL (10 pfu/punch) and 2 log10 pfu/mL (1 pfu/punch). Dauner A.L., Gilliland T.C., Jr, Mitra I., Pal S., Morrison A.C., Hontz R.D., Wu S.J. Diagnosing YF can be difficult as patients present with non-specific, flu-like symptoms that can be confused with leptospirosis, malaria, and other viral hemorrhagic fevers during differential diagnosis. FTA cards have been successfully incorporated into the molecular diagnostic protocols for many RNA viruses. Viremia/RNA in sera during WT YFV infection is transient, especially in mild or subclinical cases, and usually begins to wane 36 days after infection [1]. The .gov means its official. Following simple For punches inoculated with 1 pfu/punch, there was not a significant change in the length of detection between cards pre-incubated for one or two days with one or two desiccant packets. Purification method: Specific information to assure the reproducibility of the methods used for RNA purification. Protocols utilizing a wide variety of sample types have been optimized using FTA cards including blood, serum, tick and mosquito homogenates, dead bird impressions, throat swabs, oral fluid and epithelial suspensions [13,18,2224,3036]. Although the LOD of YFV 17D RNA on FTA punches was shown to be less than 10 pfu/mL in both PBS and flavivirus-negative human sera, the LOD of FTA cards was significantly different than the gold standard of direct RNA extraction. To assay if the reported loss in RNA yield was due to the card having absorbed moisture from the air prior to inoculation, cards were pre-incubated in the humidity chamber for one, two or three days before inoculation with YFV (illustrated in S3 Fig). The difference in LOD between one and two days of preincubation was not significant (one day vs. two days 95% CI: -0.1, 0.7); however, the difference in LOD after three days was significantly different (two days vs. three days 95% CI: 0.1, 0.9).
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