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Agency: Cordis | 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.


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.


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.


Pang Y.,National Center for Tuberculosis Control and Prevention | Lu J.,Chinese Academy of Inspection and Quarantine | Wang Y.,National Center for Tuberculosis Control and Prevention | Song Y.,National Center for Tuberculosis Control and Prevention | And 2 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2013

Rifampin (RIF) susceptibility is a key factor in determining the treatment effectiveness of the standardized treatment regimens. In Mycobacterium tuberculosis, both target gene mutation and the efflux pump play major roles in the resistance to antituberculosis drugs. By eliminating RIF-resistant strains with rpoB mutation, the choice of RIF-monoresistant strains may allow us to identify the RIF-specific efflux pump genes. This study explored the RIF monoresistance mechanism in M. tuberculosis. Data from DNA sequencing and MIC measurements revealed that specific mutations, including Ser531Leu and His526Asp in RpoB, show high-level drug resistance. Three-dimensional structure modeling provided further evidence that the affinity between RIF and RpoB mutants was in accordance with the drug resistance level of the corresponding isolates. Furthermore, transcriptionlevel analysis among the nonmutated isolates indicated that three efflux pumps (Rv0783, Rv2936, and Rv0933) might be involved in exporting RIF from the cell. Compared to 8 g/ml for wild-type Escherichia coli, the MICs for the transgenic E. coli strains with either Rv0783 or Rv2936 were 32 and 16 g/ml, respectively. In conclusion, our study indicated that several RpoB mutant types, including Ser531Leu and His526Asp, show high-level RIF resistance attributed to low affinity between RpoB mutant proteins and RIF. In addition, this work demonstrates that Rv2936 and Rv0783 may be responsible for low-level resistance to RIF by exporting RIF from cells. The predicted structure of RpoB and the newly identified efflux pumps in this study will provide a novel approach to design new drugs and develop novel diagnosis technologies. Copyright © 2013, American Society for Microbiology. All Rights Reserved.


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.

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