Unlock instant, AI-driven research and patent intelligence for your innovation.

Rapid detection method for vibrio vulnificus and vibrio harveyi

A technology of Vibrio harveyi and Vibrio vulnificus, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, and resistance to vector-borne diseases, etc. It can solve the problems of long time required for diagnosis and many detection steps

Inactive Publication Date: 2013-10-02
GUANGXI INST OF FISHERIES
View PDF1 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above detection methods all adopt modern advanced biological technology, which can accurately and conveniently diagnose the corresponding Vibrio pathogenic bacteria, but there are many detection steps and the diagnosis takes a long time

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
  • Rapid detection method for vibrio vulnificus and vibrio harveyi
  • Rapid detection method for vibrio vulnificus and vibrio harveyi
  • Rapid detection method for vibrio vulnificus and vibrio harveyi

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] (a) Bacterial liquid preparation: Inoculate the standard strain of Vibrio vulnificus, Vibrio harveyi, Streptococcus agalactiae, Streptococcus iniae, Vibrio parahaemolyticus and Photobacterium beautiful on TSA (soybean casein agar medium), Incubate at 33-35°C for 20-24 hours, and store at -4°C for later use.

[0073] (b) Sample treatment: take 0.5-1.0 g of fish tissue sample A, add 100 μl sterilized ddH 2 O, put it in a sterilized 1ml EP tube, mash the sample with a sterilized toothpick and mix it evenly, pipette the liquid to extract 20-50 μl for later use; take another bacterial liquid of standard strains (mixed Vibrio vulnificus and Vibrio harveyi) for later use;

[0074] (c) PCR amplification: Take 1.25 μl of liquid A to be tested and the standard strain as templates; add reagents in sequence: 2.5 ul 10×loading buffer, 0.25 μl Taq enzyme, 19.75 μl ddH 2 O, 0.25 μl V 1-1T , 0.25μl V 1-2 , 0.25μl , H 1-1 0.25 μl HO 1-2 Mix well with 0.5 μl 4×dNTP; place on a PCR...

Embodiment 2

[0078] (a) Bacterial liquid preparation: Inoculate the standard strain of Vibrio vulnificus, Vibrio harveyi, Streptococcus agalactiae, Streptococcus iniae, Vibrio parahaemolyticus and Photobacterium beautiful on TSA (soybean casein agar medium), Incubate at 33-35°C for 20-24 hours, and store at -4°C for later use.

[0079] (b) Sample treatment: take 0.5-1.0 g of sample B from fish liver, add 100 μl sterilized ddH 2 O, put it in a sterilized 1ml EP tube, mash the sample with a sterilized toothpick and mix it evenly, pipette the liquid to extract 20-50 μl for later use; take another bacterial liquid of standard strains (mixed Vibrio vulnificus and Vibrio harveyi) for later use;

[0080] (c) PCR amplification: Take 2.5 μl of liquid B to be tested and standard strains as templates; add reagents in sequence: 5 ul 10×loading buffer, 0.5 μl Taq enzyme, 35.5 μl ddH 2 O, 0.5 μl V 1-1T , 0.5μl V 1-2 , 0.5μl , H 1-1 0.5 μl H 1-2 Mix well with 1.0 μl 4×dNTP; place on a PCR machine ...

Embodiment 3

[0084] (a) Bacterial liquid preparation: Inoculate the standard strain of Vibrio vulnificus, Vibrio harveyi, Streptococcus agalactiae, Streptococcus iniae, Vibrio parahaemolyticus and Photobacterium beautiful on TSA (soybean casein agar medium), Incubate at 33-35°C for 20-24 hours, and store at -4°C for later use.

[0085] (b) Sample treatment: take 0.5-1.0 g of sample C from fish hepatopancreas, add 100 μl sterilized ddH 2 O, put it in a sterilized 1ml EP tube, mash the sample with a sterilized toothpick and mix it evenly, pipette the liquid to extract 20-50 μl for later use; take another bacterial liquid of standard strains (mixed Vibrio vulnificus and Vibrio harveyi) for later use;

[0086] (c) PCR amplification: Take 2.5 μl of liquid C to be tested and standard strains as templates; add reagents in sequence: 5 ul 10×loading buffer, 0.5 μl Taq enzyme, 35.5 μl ddH 2 O, 0.5 μl V 1-1T , 0.5μl V 1-2 , 0.5μl , H 1-1 0.5 μl H 1-2 Mix well with 1.0 μl 4×dNTP; place on a PCR ...

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 provides a rapid detection method for vibrio vulnificus and vibrio harveyi. The objective of the invention is to facilitate propagation and outbreak controls of the ibrio vulnificus and the vibrio harveyi in mariculture animals. According to the invention, specific primers V1 and H1 of the ibrio vulnificus and the vibrio harveyi are designed by utilization of modern advanced duplex polymerase chain reaction technology; by specific amplification of corresponding bacteria DNA fragments, and specific DNA fragment detection, the objective of rapidly, accurately, and conveniently detecting the ibrio vulnificus and the vibrio harveyi is realized. The detection method provided by the invention has advantages of strong specificity, high sensitivity, time-saving property, labor-saving property, speediness, high efficiency, and the like. Diagnosis time of pathogenic bacteria is reduced from original 3-7 days to 3-4 hours, thus improving timeliness of bacteria diagnosis. The detection method can be used for detection or disease monitoring of the ibrio vulnificus and the vibrio harveyi for marine products and aquaculture animals, and provides scientific bases for rapid diagnosis of the ibrio vulnificus and the vibrio harveyi and improves disease control capability.

Description

technical field [0001] The present invention relates to the rapid detection method of two kinds of bacteria, be specifically related to Vibrio vulnificus ( Vibrio vulnificus ) and Vibrio harveyi ( Vibrio Harvey ) detection method. Background technique [0002] Vibrio vulnificus ( Vibrio vulnificus ) and Vibrio harveyi ( Vibrio Harvey ) widely exists in the marine environment, and is a common pathogenic Vibrio in mariculture animals, mainly causing eels ( Anguilla Japonica ), grouper ( Epinephelus ), Chinese prawns ( Fenneropenaeus chinensis ), rainbow trout ( Salmo gairdneri ) and other farmed animals can cause acute sepsis and tissue function decline in seawater farmed animals. It is highly explosive, high in mortality, and has a wide range of impacts. It has brought large economic losses to seawater farmed animals. Vibrio vulnificus is also a pathogenic bacterium that is common to humans and animals. In recent years, it has caused several cases of acute septic...

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 Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCY02A50/30
Inventor 陈福艳黄婷陈晓汉杨学明梁万文程光平韦友传欧阳贤华陈明余晓丽王瑞李莉萍雷爱莹
Owner GUANGXI INST OF FISHERIES
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