Freeze-dried microchip for identifying H5 and H7 highly pathogenic variant avian influenza viruses, kit and method thereof
An avian influenza virus and microchip technology, applied in the field of molecular biology detection of viruses
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
experiment example 1
[0135] Experimental example 1 H5 and H7 subtype HA cleavage sites inserted into mutant highly pathogenic avian influenza virus freeze-dried microchip fluorescence Photo-PCR detection kit specificity verification
[0136] 1. Design primers and Taqman-MGB probes
[0137]According to the domestic detection of H5 subtype HA cleavage site insertion mutant highly pathogenic avian influenza virus and H7 subtype HA cleavage site insertion mutant highly pathogenic avian influenza virus, find out the HA gene sequence and highly pathogenic avian influenza virus The cleavage sites of H5 and H7 subtype viruses, and the HA cleavage sites of H5 and H7 subtypes are inserted into the specific conserved sequence of the HA gene of the mutant highly pathogenic avian influenza virus, and multiple pairs of primers and probes are designed. After comparison and screening, a set of optimal primers and a Taqman-MGB probe were finally determined.
[0138] H5 subtype HA cleavage site insertion varia...
experiment example 2
[0159] Experimental example 2, H5 and H7 subtype HA cleavage site insertion mutant highly pathogenic avian influenza virus freeze-dried microchip fluorescent Sensitivity Verification and Comparison of Optical PCR Detection Kit and Conventional Fluorescent RT-PCR Reagents
[0160] Set the concentration to 1 x 10 7 TCID 50 / mL of the H5N1 subtype HA cleavage site inserted into the mutant highly pathogenic avian influenza virus liquid for 10-fold serial dilution. Extract 1×10 6 TCID 50 / mL~1×10 0 TCID 50 / mL each gradient concentration of H5N1 subtype HA cleavage site was inserted into the genomic RNA of mutant highly pathogenic avian influenza virus as a template, nuclease-free water was used as a negative control, and the freeze-dried microchip fluorescent RT-PCR reagent and conventional fluorescent RT were performed -Universal sensitivity detection of PCR reagents to detect H5N1 subtype HA cleavage site insertion variant highly pathogenic avian influenza virus.
[01...
experiment example 3
[0167] Experimental example 3: H5 and H7 subtype HA cleavage sites inserted mutant highly pathogenic avian influenza virus freeze-dried microchip fluorescence Preparation and detection of photo-PCR detection kit
[0168] 1. Preparation of the kit:
[0169] The freeze-dried microchip fluorescent RT-PCR system was prepared according to Example 1.
[0170] Reagent 1: Diluent Taq Buffer (10×) (Thermo Scientific, Cat. No. B650060), 6 μl
[0171] Reagent 2: Mineral oil (Sangon Biotech, Cat. No. A630217) 1 mL
[0172] Reagent 3: positive control (H7 subtype HA cleavage site insertion variant highly pathogenic avian influenza virus, H5 subtype HA cleavage site insertion variant highly pathogenic avian influenza virus genomic cDNA mixture) 30 μl;
[0173] Reagent 4: 30 μl of negative control (no nuclease).
[0174] Reagent 5: 50 μl of nuclease-free water.
[0175] 2. Repeatability analysis of the kit
[0176] Three known positive samples were selected for intra-assay repeated ...
PUM
| Property | Measurement | Unit |
|---|---|---|
| diameter | aaaaa | aaaaa |
| PCR efficiency | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More