Rapid diagnostic kit based on loop-mediated isothermal amplification technique for hepatitis A virus genes and detection method thereof
A hepatitis A virus, ring-mediated isothermal technology, applied in the directions of microorganism-based methods, resistance to vector-borne diseases, and microbial assay/examination, which can solve problems such as genetic rapid diagnostic kits for Mycobacterium tuberculosis , to achieve the effect of significant color difference, high specificity, and rapid amplification
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment 1
[0038] The preparation of embodiment 1 kit
[0039] (1) The DNA synthesizer synthesizes the following oligodeoxynucleic acid primers:
[0040] Outer primer F3, the nucleotide sequence of which is shown in SEQ ID NO: 1;
[0041] Outer primer B3, the nucleotide sequence of which is shown in SEQ ID NO: 2;
[0042] Internal primer FIP, the nucleotide sequence of which is shown in SEQ ID NO: 3;
[0043]The nucleotide sequence of the internal primer BIP is shown in SEQ ID NO:4.
[0044] (2) Purchasing DNA polymerase: Bst DNA polymerase is placed in the container.
[0045] (3) Preparation of reaction solution: the reaction solution contains 2mmol / LdNTP, 25mmol / L Tris-Cl, 12.5mmol / L potassium chloride, 12.5mmol / L ammonium sulfate, 10mmol / L magnesium sulfate, 0.125% by volume TritonX-100, 1mol / L betaine, 2 mol / L each of the inner primers FIP / BIP and 0.25 mol / L each of the outer primers F3 / B3 were placed in the container.
[0046] (4) Purchasing reverse transcriptase: AMV reverse ...
Embodiment 2
[0062] The preparation of embodiment 2 kit
[0063] The formula of the reaction solution is: the reaction solution contains 1.6mmol / LdNTP, 20mmol / L Tris-Cl, 10mmol / L potassium chloride, 10mmol / L ammonium sulfate, 8mmol / L magnesium sulfate, 0.1vol%TritonX-100, 0.8mol / L betaine, inner primer FIP / BIP each 1.6mol / L and outer primer F3 / B3 each 0.2mol / L.
[0064] The chromogenic solution is EvaGreen.
[0065] Others are the same as embodiment 1.
Embodiment 3
[0066] Example 3 Application of Hepatitis A Virus Gene Rapid Diagnosis Kit
[0067] Collect samples according to the method in 2.1 of GB / T 18936-2003, add 175mL glycine buffer solution (pH 7.5, glycine 0.5mol / L, NaCL 0.3mol / L) to 25g shellfish sample, mix well with agitator, 37℃ 30min. 4°C 1000r / m20min, take the supernatant. Use 8% PEG8000 (with a final NaCl concentration of 0.75 mol / L) to settle the virus overnight at 4°C. 10000r / m15min, discard the supernatant, resuspend the pellet with 10mL PBS buffer (pH7.4), add an equal volume of 3-chloro-3-fluoro-ethane, mix well, 10000r / m15min at 4°C, carefully absorb the supernatant, The virus was sedimented overnight at 4° C. with 8% PEG8000 (with a final NaCl concentration of 0.75 mol / L). 14000r / m15min at 4°C, discard the supernatant, and resuspend the pellet in an appropriate amount of PBS buffer. Take 1 / 4 volume of the suspension, extract it once with an equal volume of chloroform, and carefully collect the supernatant for RNA...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com