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Duck plague (DP) caused by duck enteritis virus (DEV) is an acute infection of waterfowl. Apart from geese and ducks free-ranging swans are susceptible to infection. The exact epidemiological situation in relation to DEV infection in Eastern Europe is unknown. Therefore it was reasonable to conduct a study on DEV occurrence using the modern but simple loop-mediated isothermal amplification (LAMP). The use of LAMP facilitated DEV DNA detection in a time below 30 min. The method was also specific solely for DEV, while its detection limit was 1 pg of DNA. The aim of this study was the optimization of LAMP for detection of DEV infections among free-ranging water birds. The presence of DEV DNA was identified in 96 birds (72.7%), predominantly in wild ducks (Anas platyrhynchos) and mute swans (Cygnus olor). The results were recorded in visible light as a green color of reaction mixtures and their fluorescence under ultraviolet light. The presented study indicates the usefulness of the developed LAMP technique for DEV detection and the high percentage of free-ranging water birds infected with this virus. The study will be continued in the future for the determination of DEV occurrence in domestic geese and ducks.
The study describes construction of five recombinant very virulent (vv) and very virulent plus (vv+) strains lacking meq and viral telomerase (vTR). Deletion of both copies of meq and vTR was achieved by Red E/T recombination in GS1783 E. coli cells. The constructed bacterial artificial chromosome (BAC) clones reconstituted in chicken embryo fibroblasts were examined by immunofluorescence assay to compare the features of recombinant strains with wild-type viruses. The results demonstrated that recombinant BAC strains caused slightly reduced cytophatic effect and decreased level of the fluorescence obtained from the monoclonal antibody in comparison to the parental viruses. Generation of recombinant BAC clones may provide more detailed information on the function of Marek's disease virus oncogenes and the potential use of recombinants for the preparation of the new vaccine against Marek's disease.
DNA microarrays is a method used for the determination of gene expression level, detection of single nucleotide polymorphism (SNP) and quantification of the exact number of viral gene copies per one cell. The technique has a broad application in studies on tumorous diseases of humans and animals. Microarrays are also applied in studies of drug influence on the cell metabolism and the molecular silencing of genes. The main advantage of DNA and protein microarrays is the possibility to analyse a number of complex cellular processes in progress of a tumoral disease during a single run of analysis. In the case of a tumorous disease the microarray technique makes it possible to quickly identify the main genes responsible for the oncogenesis and to determine the type of the disease. In spite of a high costs of microarray production as well as the need to synthesize probes and substrates for specific analyses, microarrays become increasingly popular in numerous research laboratories.
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