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Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: KBBE-2009-1-4-01 | Award Amount: 3.99M | Year: 2010

Detection methods are the first tools used by national plant protection organisations (NPPO) and inspection services in order to find incursions of quarantine plant pathogens or pests (Q-pests) across a border, a crucial step to implement Council Directive 2000/29/EC. This is often done visually in the first instance, with support from a laboratory for confirmatory testing and subsequent monitoring. Reliance on laboratory testing causes significant delays when action is only taken on the return of results from the laboratory to which the samples were sent. Thus, there is a real need for rapid, simple and robust detection methods that can be deployed by NPPOs in the field with inspection services to enable early detection of Q-pests. The Q-detect consortium aims to develop detection methods based on biochemical (detecting volatile organic compounds [VOC] and nucleic acid), acoustic (including resonance), remote imaging (incorporating spectral and automated data analysis) and pest trapping (insect pests and pathogen vectors) techniques. The careful selection of traded products (primarily potato and forestry/trees) ensures the methods will be developed on high priority targets for the EU such as the pine wood nematode (Bursaphelenchus xylophilus), potato brown rot (Ralstonia solanacearum) and potato ring rot (Clavibacter michiganensis ssp. Sepedonicus), Asian longhorn beetle (Anoplophora glabripennis) and a range of whitefly transmitted viruses. The diversity of targets enables the Q-detect consortium to work on suites of complementary techniques; this is of particular importance since the diverse range of targets listed in Directive 2000/29/EC means no single detection method will be suitable for all Q-pests. Critically, NPPOs and third country institutes are partners, which will enable testing, and validation of methods at real outbreak sites where these are absent in the EU. SME partners ensure access to technology and routes for exploitation after the project ends.


Grant
Agency: European Commission | Branch: FP7 | Program: CP-FP | Phase: KBBE-2008-1-4-01 | Award Amount: 4.14M | Year: 2009

Development of accurate identification tools for plant pathogens and pests is vital to support European Plant Health Policies. For this project Council Directive 2000/29/EC is important, listing some 275 organisms for which protective measures against introduction into and their spread within the Community needs to be taken. Those threats are now greater than ever because of the increases in the volumes, commodity types and origins of trade, the introduction of new crops, the continued expansion of the EU and the impact of climate change. Currently identifying pathogens (in particular new emerging diseases) requires a staff with specialised skills in all disciplines (mycology, bacteriology, etc.); which is only possible within big centralised laboratory facilities. Taxonomy, phytopathology and other fields which are vital for sustaining sound public policy on phytosanitary issues are threatened with extinction. Modern molecular identification/detection techniques may tackle the decline in skills since they often require much less specialist skills to perform, are more amenable for routine purposes and can be used for a whole range of different target organisms. Recently DNA barcoding has arisen as a robust and standardised approach to species identification. QBOL wants now to make DNA barcoding available for plant health diagnostics and to focus on strengthening the link between traditional and molecular taxonomy as a sustainable diagnostic resource. Within QBOL collections harbouring plantpathogenic Q-organisms will be made available. Informative genes from selected species on the EU Directive and EPPO lists will be DNA barcoded from vouchered specimens. The sequences, together with taxonomic features, will be included in a new internet-based database system. A validation procedure on developed protocols and the database will be undertaken across worldwide partners to ensure robustness of procedures for use in a distributed network of laboratories across Europe


Xi G.,Japan International Center for Materials Nanoarchitectonics | Xi G.,Chinese Academy of Inspection and Quarantine | Ye J.,Japan International Center for Materials Nanoarchitectonics | Ye J.,Tianjin University | And 4 more authors.
Journal of the American Chemical Society | Year: 2012

Metal/semiconductor hybrid materials of various sizes and morphologies have many applications in areas such as catalysis and sensing. Various organic agents are necessary to stabilize metal nanoparticles during synthesis, which leads to a layer of organic compounds present at the interfaces between the metal particles and the semiconductor supports. Generally, high-temperature oxidative treatment is used to remove the organics, which can extensively change the size and morphology of the particles, in turn altering their activity. Here we report a facile method for direct growth of noble-metal particles on WO 3 through an in situ redox reaction between weakly reductive WO 2.72 and oxidative metal salts in aqueous solution. This synthetic strategy has the advantages that it takes place in one step and requires no foreign reducing agents, stabilizing agents, or pretreatment of the precursors, making it a practical method for the controlled synthesis of metal/semiconductor hybrid nanomaterials. This synthetic method may open up a new way to develop metal-nanoparticle-loaded semiconductor composites. © 2012 American Chemical Society.


Jin B.,Shenzhen Entry Exit Inspection and Quarantine Bureau | Xie L.,Shenzhen Entry Exit Inspection and Quarantine Bureau | Guo Y.,Shenzhen Entry Exit Inspection and Quarantine Bureau | Pang G.,Chinese Academy of Inspection and Quarantine
Food Research International | Year: 2012

The extensive use of pesticides in modern farming on fruit and vegetables has posed risks to public health and environment. In this study, the methods for extraction and detection of pesticides in juice and fruit wine were reviewed. Sample preparation is an important step in the analytical method, and the advantages of various new extraction techniques over the classical solvent extraction have been highlighted. Current methods involve the use of one or the combination of some of the following techniques for both the sample extraction and clean-up steps: liquid-liquid extraction, solid-phase extraction, solid-phase microextraction, stir bar sorptive extraction, matrix solid-phase dispersion and single-drop microextraction. Determination of low-level pesticide residues in juice and wine has been performed mainly by chromatographic methods employing selective detectors or, in an increasing proportion, coupled to mass spectrometry for the quantification and simultaneous identification of residues. © 2011 Elsevier Ltd.


Wang J.,Beijing University of Chemical Technology | Du Z.,Beijing University of Chemical Technology | Yu W.,Chinese Academy of Inspection and Quarantine | Qu S.,Beijing University of Chemical Technology
Journal of Chromatography A | Year: 2012

A liquid-phase microextraction (LPME) methodology based on the use of porous polyvinylidene fluoride (PVDF) hollow fibres was developed for extracting seven pesticides from cucumbers. The seven pesticides include propoxur, carbofuran, atrazine, cyanatryn, metolachlor, prometryn and tebuconazole. The PVDF hollow fibre provides higher extraction efficiency due to its higher porosity and better solvent compatibility. A new desorption methodology was developed since some pesticides were absorbed by the wall pore of the PVDF. Ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used for pesticide analysis. In order to obtain high recoveries and enrichment factors of the analytes, several parameters such as method of sealing, acceptor phase (organic solvents), stirring speed, extraction time, salting out effect, desorption mode and time were optimized. A fast, simple method for closing fibre ends was practiced by using mechanical crimping. Pesticides were extracted from the sample to the organic solvent and then desorbed in a mixture of methanol:water (1:1 v/v) prior to chromatographic analysis. Limits of detection (LOD) for the multi-reaction-monitoring (MRM) mode of the method varies from 0.01 to 0.31μg/kg with optimized sample preparation. Calibration curves are linear with R2≥0.991. Enrichment factor of the hollow fibre LPME ranges from 100 to 147. Matrix effect has been considered and is in the range of 76-122%. The relative recoveries from cucumber samples are between 63% and 119% with the relative standard deviation (RSD, n=6) lower than 20%. © 2012 Elsevier B.V.


Pan X.,Chinese Academy of Inspection and Quarantine
BMC Ecology | Year: 2016

Background: Species-area relationship (SAR), endemics-area relationship (EAR) and overlap-area relationship (OAR) are three important concepts in biodiversity study. The application of fundamental equations linking the SAR, EAR and OAR, can enrich the axiomatic framework of the species-area theory and deepen our understanding of the mechanisms of community assembly. Results: Two fundamental equations are derived and extended to power law model and random replacement model of species-area distribution. Several important parameters, including the overlap index and extinction rate, are defined and expressed to enrich the species-area theory. For power law model, both EAR and OAR have three parameters, with one more parameter of the total area than SAR does. The EAR equation is a monotonically increasing function for parameter c and z, and a monotonically decreasing function for parameter A. The extinction rate, with two parameters, is a monotonically increasing function for parameter z, and a monotonically decreasing function for parameter A. The overlap index is a monotonically increasing function for parameter A, and a monotonically decreasing function for parameter z, independent of parameter c. Conclusions: The general formats of SAR, EAR, OAR, overlap index, overlap rate, sampling rate and extinction rate, are derived and extended to power law model and random replacement model as the axiomatic framework of species-area theory. In addition, if the total area is underestimated, the extinction rate will be overestimated. © 2016 The Author(s).


Yang Y.,Chinese Academy of Inspection and Quarantine
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology | Year: 2012

Marburg virus and Ebola virus are acute infections with high case fatality rates. A rapid, sensitive detection method was established to detect Marburg virus and Ebola virus by multiplex real-time fluorescence quantitative PCR. Designing primers and Taqman probes from highly conserved sequences of Marburg virus and Ebola virus through whole genome sequences alignment, Taqman probes labeled by FAM and Texas Red, the sensitivity of the multiplex real-time quantitative PCR assay was optimized by evaluating the different concentrations of primers and Probes. We have developed a real-time PCR method with the sensitivity of 30.5 copies/microl for Marburg virus positive plasmid and 28.6 copies/microl for Ebola virus positive plasmids, Japanese encephalitis virus, Yellow fever virus, Dengue virus were using to examine the specificity. The Multiplex real-time PCR assays provide a sensitive, reliable and efficient method to detect Marburg virus and Ebola virus simultaneously.


Jiang Y.L.,Chinese Academy of Inspection and Quarantine
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] | Year: 2011

A virus was isolated from cultured sick giant salmander (Andrias davidianus ) in a farm, Shanxi Province, China. Skin ulceration and necrosis of the distal limbs are main clinical symptoms. Virus propagated and caused CPE at 10 degrees C to 30 degrees C in BF-2, CO, CHSE, FHM cells. The optimum condition of replication was in BF-2 cells at 25 degrees C. The virus was proved to be senstive to chloroform, heat, pH3 and pH10 treatment. Viral replication was inhibited by 5-Fluoro-2-deoxyuridine (FUDR). These results indicated that the virus possessed an envelope and DNA as the genome. Electron-microscopic observation of thin-section showed numerous hexagonal viral particles measuring 130 nm to 150 nm in diameter orderly arranged in a lattice form in cytoplasm of BF-2 cells. The particles showed typical iridovirus morphology. A 413 bp fragment was amplified from the viral main capsid protein gene by PCR. The fragments was sequenced and analysed. The results showed the isolate shared more than 96% nucleotide identity with some Ranaviruses. We suggested that this virus was named as Andrias davidianus iridovirus (ADIV) tentatively.


Cheng Y.,Chinese Academy of Inspection and Quarantine | Dong Y.,Chinese Academy of Inspection and Quarantine
Food Control | Year: 2011

A novel fast ultra-sensitive approach was developed herein to identify melamine contaminant in eggs using portable compact surface-enhanced Raman Spectroscopy (SERS) on gold nanosubstrates. Trace amounts of melamine spiked in albumen and yolk were characterized and quantified by SERS with partial least squares (PLS) analysis, good models were acquired for melamine in albumen at concentration range of 2.5-100 mg/kg with the determination coefficient of actual concentration versus predicted concentration R 2 = 0.94, the root mean standard error of cross validation RMSECV = 12.38, the limit of detection (LOD) 1.1 mg/kg, and in yolk 5.0-200 mg/kg with R 2 = 0.98, RMSECV = 12.14, LOD = 2.1 mg/kg, respectively. The assay was rapid in less than 30 min, and the detection was sensitive. As a fast screening scheme, the method could be suitably applied to the domain of on-site egg products quality control and market surveillance thereof. © 2010 Elsevier Ltd.


NS1 of influenza A virus is a key multifunctional protein that plays various roles in regulating viral replication mechanisms, host innate/adaptive immune responses, and cellular signalling pathways. These functions rely on its ability to participate in a multitude of protein-protein and protein-RNA interactions. To gain further insight into the role of NS1, a tandem affinity purification (TAP) method was utilized to find unknown interaction partner of NS1. The protein complexes of NS1 and its interacting partner were purified from A549 cell using TAP-tagged NS1 as bait, and co-purified cellular factors were identified by mass spectrometry (MS). We identified cellular β-tubulin as a novel interaction partner of NS1. The RNA-binding domain of NS1 interacts with β-tubulin through its RNA-binding domain, as judged by a glutathione S-transferase (GST) pull-down assay with the GST-fused functional domains of NS1. Immunofluorescence analysis further revealed that NS1 with β-tubulin co-localized in the nucleus. In addition, the disruption of the microtubule network and apoptosis were also observed on NS1-transfected A549 cells. Our findings suggest that influenza A virus may utilize its NS1 protein to interact with cellular β-tubulin to further disrupt normal cell division and induce apoptosis. Future work will illustrate whether this interaction is uniquely specific to the 2009 pandemic H1N1 virus.

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