WO2024136540A1 - Composition pour la détection de la virémie printanière de la carpe et procédé de détection à cet effet - Google Patents
Composition pour la détection de la virémie printanière de la carpe et procédé de détection à cet effet Download PDFInfo
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- WO2024136540A1 WO2024136540A1 PCT/KR2023/021304 KR2023021304W WO2024136540A1 WO 2024136540 A1 WO2024136540 A1 WO 2024136540A1 KR 2023021304 W KR2023021304 W KR 2023021304W WO 2024136540 A1 WO2024136540 A1 WO 2024136540A1
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- carp
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- seq
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- coding gene
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Images
Definitions
- RT-PCR reverse-transcription PCR
- SVC Aphanomyces invadans
- FISH fluorescent peptide nucleic acid in-situ hybridisation
- the present invention includes a primer pair for amplifying the Matrix protein coding gene and/or the Nucleocapsid coding gene of Spring viraemia of carp (SVC), It relates to a composition for detecting carp spring virus disease caused by carp spring virus infection.
- the amount of the amplification product can be detected by a fluorescence signal.
- the intercalating method uses an intercalator that binds to the double-stranded DNA of the amplification product to which the probe is bound and displays fluorescence.
- the 5' end is labeled with a fluorescent substance and the 3' end is labeled with a quencher.
- the present invention also relates to carp caused by Spring viraemia of carp infection, comprising a primer pair for amplifying the Matrix protein coding gene or the Nucleocapsid coding gene of Spring viraemia of carp.
- This relates to a kit for detecting spring virus disease (Spring viraemia of carp; SVC).
- biological samples analyzed include body fluid samples, including blood, serum, plasma, lymph, breast milk, urine, feces, ocular fluid, saliva, semen, brain extracts (e.g., brain pulverized material), spinal fluid, appendix, and spleen. and tonsil tissue extract, but are not limited thereto.
- body fluid samples including blood, serum, plasma, lymph, breast milk, urine, feces, ocular fluid, saliva, semen, brain extracts (e.g., brain pulverized material), spinal fluid, appendix, and spleen. and tonsil tissue extract, but are not limited thereto.
- RNA sample of Spring viraemia of carp the cause of carp spring virus disease (Spring viraemia of carp; SVC), and analyzed to determine the causative virus. Detection was performed.
- PCR was performed using Exicycler96TM (Bioneer, Korea).
- Table 2 The composition of the reactants for PCR targeting the matrix protein coding gene is shown in Table 2, and the composition of the reactant for PCR targeting the matrix protein coding gene and the nucleocapsid coding gene is shown in Table 3.
- real-time PCR was performed by creating a PCR master mix and adding 5 ⁇ l viral RNA.
- Table 4 shows the amplification conditions and hybridization reaction conditions of the reactants, and shows the process of amplifying and hybridizing the viral RNA sample and then increasing the temperature of the hybridized product. The above process was performed by real-time PCR.
- the causative agent of spring viraemia of carp (SVC) and Real-time PCR was performed by adding 5 ⁇ L of nucleic acid of similar viruses.
- Figures 2a and 2b show the results of PCR using a primer pair with improved sensitivity and a TaqMan probe for the nucleic acid of a virus similar to the causative agent of spring viraemia of carp (SVC).
- compositions, kit, and detection method that can sensitively molecularly diagnose spring viraemia of carp, the causative agent of carp spring virus disease, through the primer pair according to the present invention, it is simple to determine whether the pathogen is infected. ⁇ It is effective in detecting quickly and accurately.
Abstract
La présente invention concerne une composition pour détecter la virémie printanière de la carpe, et un procédé de détection associé, et plus particulièrement : une paire d'amorces pour amplifier les gènes codant pour une protéine de matrice ou une nucléocapside de la virémie printanière de la carpe ; une composition et un kit pour détecter la virémie printanière de la carpe, comprenant chacun cette composition ; et un procédé pour détecter la virémie printanière de la carpe à l'aide de cette composition.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2022-0180628 | 2022-12-21 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2024136540A1 true WO2024136540A1 (fr) | 2024-06-27 |
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