Check patentability & draft patents in minutes with Patsnap Eureka AI!

SVCV (spring viraemia of carp virus) standard sample and preparation method thereof

The technology of spring carp virus and standard sample is applied in the field of standard sample of carp spring virus and its preparation, which can solve the problems of imperfect epidemic prevention system, disease of aquatic animals, and difficulty in obtaining viruses.

Active Publication Date: 2015-07-22
吴斌 +3
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the deterioration of the breeding ecological environment, poor awareness of disease prevention, widespread drug abuse, poor quality of aquatic animal seedlings, and unsound epidemic prevention systems, etc., aquatic animals have disease
The fish samples sent for inspection in the system are usually relatively healthy visually, but carp spring virus usually only develops at a suitable temperature, and the current PCR method can only be detected from fish with severe infection showing clinical symptoms Viruses, potential viruses cannot be detected
With the increase of import and export trade, the time limit of the inspection process has been shortened. Most of the fish diseases need to go to the cells to proliferate first, and it is difficult for ordinary aquatic animal disease laboratories to obtain the virus, which brings great impact on the quality control and research of the laboratory great challenge

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
  • SVCV (spring viraemia of carp virus) standard sample and preparation method thereof
  • SVCV (spring viraemia of carp virus) standard sample and preparation method thereof
  • SVCV (spring viraemia of carp virus) standard sample and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Preparation of carp spring virus standard sample

[0025] 1. Virus identification

[0026] According to OIE and "GB / T15805.5-2008 Fish Quarantine Methods Part 5: Carp Spring Virus (SVCV)" method, the fish disease materials collected daily by Liaoning Entry-Exit Inspection and Quarantine Bureau (the virus strain comes from samples) Perform pretreatment and RT-PCR detection.

[0027] RNA extraction

[0028] ① Trizol method to extract viral RNA:

[0029] ②Take three 1.5mL sterilized EP tubes, two virus samples and one negative control, and number each tube.

[0030] ③ Add 600 μL of lysate and 200 μL of sample to each tube; then add 200 μL of chloroform, shake and mix for 5 seconds on the mixer. Centrifuge at 12000r / min for 15min at 4°C.

[0031] ④ Take the same number of EP tubes, add 500 μL of isopropanol pre-cooled at -20°C, and number each tube. Take at least 500 μL of supernatant after centrifugation and mix by inversion. Centrifuge at 12000r / min for 1...

Embodiment 2

[0065] Embodiment 2. The preparation of standard sample

[0066] 1. Cell recovery and virus proliferation

[0067] 1. Recovery of EPC cells

[0068] ①Take out the cryopreservation tube from the liquid nitrogen, and quickly put it into a water bath at 37-38°C to make it melt rapidly. Then centrifuge at 1000r / min for 5min.

[0069] ② Aspirate the supernatant, add 1mL of M199 culture medium, and mix by pipetting. Transfer to a 25cm2 culture flask, and continue to add culture solution to a final volume of 6mL.

[0070] ③Mark the cell name and date.

[0071] ④Microscopic examination.

[0072] ⑤Put it in a low-temperature incubator at 20°C for cultivation.

[0073] 2. Proliferation of virus

[0074] ①Choose 4 bottles of EPC cells covered with a single layer, and discard the culture medium. The tissue disease materials obtained in Example 1 were inoculated 24 hours after the cells were subcultured and preserved by the laboratory, of which 1 bottle was used as a control group ...

Embodiment 3

[0098] Embodiment 3 standard sample homogeneity and stability test

[0099] The uniformity of the standard sample is the basic property of the standard sample. Its purpose is to explain whether the characteristic value is uniform among various parts through the uniformity test on the one hand, and to understand the degree of non-uniformity of the characteristic value among different parts on the other hand. Judge whether the degree of inhomogeneity is acceptable and whether the standard sample can be used. Regardless of whether the initial uniformity test has been carried out during the preparation of standard samples, all standard samples prepared in batches and packaged into the smallest packaging unit must be tested for uniformity. Uniformity inspection is also required when bulk packaging is divided into the smallest packaging unit. This is an indispensable procedure in the process of preparing standard samples, and it is also the most basic condition to ensure accurate d...

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

No PUM Login to View More

Abstract

The invention discloses a preparation method of an SVCV (spring viraemia of carp virus) standard sample. The preparation method takes a virus strain SVCV-DL to prepare the standard sample and comprises the following steps: identifying the SVCV-DL and preparing the standard sample, namely propagating the virus strain to obtain a virus culture material, and carrying out a specific freeze-drying process with a freeze-drying protecting agent prepared from mycose, proline, skimmed milk powder and de-ionized water to obtain the standard sample. The standard sample is successfully developed after being subjected to operations of uniformity and stability inspection, qualitative inspection, value fixation and the like finally. The standard sample not only meets the requirement of detection research, medicine research and application research on standard samples, but also provides a technical support for technical guidance of inspection and quarantine institutions and service export enterprises.

Description

technical field [0001] The invention belongs to the technical field of virus culture standard samples for detection and preparation methods thereof, and in particular relates to carp spring virus standard samples and preparation methods thereof. Background technique [0002] my country is the world's largest producer and exporter of aquatic products. At present, the total amount of aquatic products in my country has accounted for 40% of the total global fishery output, ranking first in the world for 15 consecutive years, and the output of aquaculture aquatic products accounts for 70% of the world's total aquaculture output. Aquatic products have played an important role in ensuring national food security, increasing farmers' income and stabilizing the rural economy. The barriers to the export of aquatic products in my country are mainly green barriers, such as Japan's "Positive List System", the European Union's "EU Food and Feed Safety Management Regulations" and related la...

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
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N7/00C12R1/93
CPCC12Q1/02
Inventor 吴斌肇慧君蔡晓萍张雪
Owner 吴斌
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com