Compositions for detecting of influenza viruses and kits and methods using same
A technology of influenza virus and composition, which is applied in the field of detection of novel composition of influenza virus, and can solve problems such as indistinguishability
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Embodiment 1
[0216] Exemplary substrate and separation system of the present invention
[0217] figure 1 It is a schematic representation of the structure of the substrate of the present invention. The substrate consists of 3 parts: X, Y and Z. One end of the core molecule or molecular complex (fragment Y) with a specific cleavage site is connected to the tag molecule (X), and the purpose of the tag molecule is to detect the cleaved substrate. The other end is connected to a structural fragment (Z) that separates the processed substrate from the unprocessed substrate. During the cleavage of molecule Y, the substrate is divided into two parts: (1) a tag fragment (TS) containing part X and part Y and (2) a separated fragment containing part Y and Z (SS).
[0218] in figure 1 When the substrate described in is cleaved by neuraminidase, the Z fragment (isolated fragment) is used to separate the processed substrate from the unprocessed substrate. Therefore, only the TS (containing the tag molecu...
Embodiment 2
[0225] Neuraminidase detection based on beads and fluorescence (FRET)
[0226] This example describes the detection of the presence of avian influenza H5N1 virus and other types of influenza in clinical samples. The influenza virus family uses a combination of specific hemagglutinin and neuraminidase pairs. The pair showed specific cleavage activity. In this embodiment, the substrate is composed of hemagglutinin (HA) (or a part thereof) and a fluorophore with one end connected to a colored bead (corresponding to Z). The fluorophore can also be attached to the HA moiety. The HA sialic acid receptor is used to connect the HA moiety to one end, and the molecule containing the sialic acid and the quencher to the other end. An exemplary sialic acid part is described in Figure 5 in. The other end of the HA molecule ends with the sialic acid receptor, which uses receptor-substrate (natural substrate mimicking neuraminidase) affinity and quencher as its The sialic acid (corresponding to ...
Embodiment 3
[0231] Matrix determination of influenza type used to detect unknown types of influenza virus (general description)
[0232] The influenza virus is defined according to the combination of neuraminidase and HA virus protein. The name of the virus is defined according to this combination, that is, the H5N1 virus has a combination of HA5 and neuraminidase 1 on its spikes. Due to multiple mutations and evolutions in the virus, the virus will evolve different subtypes in each season. Due to global concerns that one type of mutation may be particularly aggressive (as happened during the Spanish flu in 1918, which killed more than 25 million people in Europe), there is an urgent need for rapid detection and isolation. In order to monitor the threat of epidemic spread caused by these viruses and minimize them. It is currently not possible to determine the specific spectrum of a particular flu type in a certain flu season in a rapid test. The method described in the present invention enabl...
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