Team:Tokyo Tech
Project Description
Dengue Virus Detection System with multiple colors
Mechanism
For 2018 project, we try to develop mosaic-envelope virus that can detect the presence of antibodies in serum that correspond to specific type of virus. This mosaic-envelope virus can detect all the serotypes of dengue viruses and can contribute to versatile dengue virus detection. The experimental procedures are categorized as the following three steps.
STEP1: We construct integrated plasmid that codes essential viral proteins (both structural and non-structural) and green fluorescence protein. (Note: If the size is too large, they will be separated.) The envelope (E) genes are derived from 2 or more dengue serotypes so that envelope proteins can be mixed to detect multiple types of antibodies.
STEP2: We transduce the plasmid into suitable eukaryotic cells and culture the cells which are successfully transduced. (Drug selection and a series of passage may be needed.) Most of the transduced cells are expected to produce mosaic-envelope virus.
STEP3: We culture the target mammalian cells that are for checking the infection by mosaic-envelope virus. After mixing the cells and the virus, we measure the fluorescence intensity to check whether the mosaic-envelope virus infected and GFPs are produced or not. As control experiment, we add antibodies that correspond to one of the dengue serotypes to check whether the function of the virus is inhibited or not.
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Contents
Project Description
Dengue Virus Detection System with multiple colors
Mechanism
Virus detection system with multi-colors
For 2018 project, we try to develop mosaic-envelope virus that can detect the presence of antibodies in serum that correspond to specific type of virus. This mosaic-envelope virus can detect all the serotypes of dengue viruses and can contribute to versatile dengue virus detection. The experimental procedures are categorized as the following three steps.
STEP1: We construct integrated plasmid that codes essential viral proteins (both structural and non-structural) and green fluorescence protein. (Note: If the size is too large, they will be separated.) The envelope (E) genes are derived from 2 or more dengue serotypes so that envelope proteins can be mixed to detect multiple types of antibodies.
STEP2: We transduce the plasmid into suitable eukaryotic cells and culture the cells which are successfully transduced. (Drug selection and a series of passage may be needed.) Most of the transduced cells are expected to produce mosaic-envelope virus.
<p style="font-size:16px;font-size: 16px; text-indent:1em;padding-top: 15px"> STEP3: We culture the target mammalian cells that are for checking the infection by mosaic-envelope virus. After mixing the cells and the virus, we measure the fluorescence intensity to check whether the mosaic-envelope virus infected and GFPs are produced or not. As control experiment, we add antibodies that correspond to one of the dengue serotypes to check whether the function of the virus is inhibited or not.
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<figure>
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<figcaption style="font-size: 16px">Fig. 1 Basic Flow of dengue virus detection system</figcaption>
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