VHSV (viral haemorrhagic septicaemia virus) visual nucleic acid strip detection primer group and detection method
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A technology of viral hemorrhage and nucleic acid test strips, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve problems such as inapplicability of rapid differential diagnosis, difficulty in satisfying rapid detection, time-consuming and laborious detection, etc., to achieve Improve identification work efficiency, low application cost, and intuitive results
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
The result of cell culture isolation VHSV method is reliable, but the detection cycle is long, which is not suitable for rapid differential diagnosis; the traditional virus cell culture method and serological method are limited by their own characteristics, the detection is time-consuming and laborious, and it is difficult to meet the needs of rapid detection
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0036] Embodiment 1: VHSV test strip detection
[0037] PCR product test strip detection test
[0038] Take 5 μL of the amplification product and drop it on the sample pad of the test strip, and add 3 drops of buffer solution at the same time, observe the quality control line and detection line after 10 minutes, and judge the negative and positive results.
[0039] The amplified product is dropped on the sample pad of the test strip for detection, and a red band appears in the detection band area, and there is no red band in the blank control ( figure 1 ).
Embodiment 2
[0040] Embodiment 2: the specificity of nucleic acid test strip method
[0041] VHSV281-F and VHSV281-R primers were used for RT-PCR amplification of VHSV, IHNV, and SVCV RNA and nucleic acid test strip detection, as can be seen from the detection results ( figure 2 ), only VHSV amplified products had specific red bands, and IHNV, SVCV and blank control had no bands at the detection line position, showing the good specificity of the detection method.
Embodiment 3
[0042] Embodiment 3: the sensitivity of nucleic acid test strip method
[0043] Extract and determine the concentration of VHSV pseudovirus RNA, respectively diluted to 6.6, 33, 66, 6.6 × 10 2 , 6.6×10 3 , 6.6×10 4 , 6.6×10 5 , 6.6×10 6 , 6.6×10 7 Copy / μL, carry out RT-PCR, detect with nucleic acid test strips, it can be seen from the results ( image 3 ), the detection limit of the nucleic acid test strip method can reach 33 copies / μL.
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
PUM
Login to view more
Abstract
The invention discloses a VHSV visual nucleic acid strip detection primer group and detection method. The sequences of primers are VHSV281-F:5'-FITC-CAGGCGTTGTCCGTGCTTCT-3', 5' terminal marking FITC and VHSV281-R:5'-biotin-TCCCCGAGTTTCTTGGTGATGTA-3',5' terminal marking biotin. The detection method comprises the following steps of (1) RT-PCR amplification; (2) PCR product test strip detection test. The method has the advantages that the operation is convenient, the specificity and the sensitivity are high, the result is intuitive, stable, reliable and environment-friendly and the like and is suitable for VHSV rapid screening of the port quarantine laboratory, the identification working efficiency of the entry and exit port is improved, the requirement for fast customs clearance on import and export trades can be well met, a used RT-PCR product detection test strip is a general test strip and can be widely used for development of PCR diagnostic reagents, no instruments and equipment such as an electrophoresis apparatus and a transilluminator are used in the detection method, the application cost is low, and the method has a great promoting effect on popularization and application of the PCR detection method to base laboratories.
Description
Technical field: [0001] The invention relates to a primer set and a detection method for visual nucleic acid test strip detection of viral hemorrhagic sepsis virus (VHSV). Background technique: [0002] Viral hemorrhaic septicaemia (viralhaemorrhaic septicaemia, VHS) is a severe infectious disease caused by rhabdoviruses in dozens of species of fish such as salmon, trout, pike and turbot. ~100%. The disease is mainly prevalent in the northern hemisphere, and it is a disease required by the World Organization for Animal Health (OIE) to be declared. my country lists it as a second-class disease. The VHSV genome is a single-stranded negative-strand RNA whose linear genome encodes five proteins, including: nucleocapsid protein (N protein), two structural proteins (M1 and M2 proteins), glycoprotein (G protein) and RNA polymerase (L protein). VHSV particles are about 180nm long and 70nm wide. Its genome is about 12k bases in length. [0003] At present, the commonly used meth...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more