Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Reagent kit for fast detection of vibrio proteolyticus and its detection process

A technology for Vibrio proteolyticus and kits, which are applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc., can solve problems such as cumbersome and time-consuming, and achieve high sensitivity, convenient detection, and good specificity. Effect

Inactive Publication Date: 2009-11-04
JINAN UNIVERSITY
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the techniques for identification of Vibrio include: 1. Classical physiological and biochemical identification methods, which are cumbersome, time-consuming, and require operators to have good professionalism; 2. Bacteria rapid automatic identification systems, which require expensive instruments and equipment and Skilled operators; 3. Nucleic acid-based detection technology, including DNA probe and PCR technology (polymerase chain reaction technology), has the characteristics of fast, simple and sensitive compared with the above-mentioned detection technology, but has not yet See Patents for PCR Technology and Detection Kit for Detection of Vibrio Proteolyticus

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
  • Reagent kit for fast detection of vibrio proteolyticus and its detection process
  • Reagent kit for fast detection of vibrio proteolyticus and its detection process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 A kit for rapid detection of Vibrio proteolyticus was made according to the following formula:

[0034] (a) Reagent A (enrichment medium): contains tryptone, beef extract, yeast extract and agar powder in a weight ratio of 5:3:1:18.

[0035] (b) Polymerase chain reaction (25 μl system) reagents: including reaction master mix reagent B and reagent C, reagent B contains 10 times polymerase chain reaction buffer (containing MgCl 2 15mM), specific primer pair VpS and VpA, dNTP (a mixture of four deoxynucleotides), bovine serum albumin and sterile double distilled water, the contents are 2.5μl, 0.5μl (10μM), 0.5μl (10μM) , 0.5 μl (10 μM each), 0.5 μl (5 mg / ml) and 19.375 μl.

[0036] Reagent C: Taq enzyme, 0.125 μl (5 units / μl).

[0037] (c) Electrophoresis and chromogenic reagents, including reagent D, reagent E and reagent F.

[0038] Reagent D: 50 times TAE (electrophoresis buffer), containing trihydroxymethylaminomethane-acetic acid and ethylenediaminetetraa...

Embodiment 2

[0046] Example 2 A kit for rapid detection of Vibrio proteolyticus was made according to the following formula:

[0047](a) Reagent A (enrichment medium): containing tryptone, beef extract, yeast extract and agar powder in a weight ratio of 5:3:1:18.

[0048] (b) Reagents for polymerase chain reaction (50 μl system): including reaction premix reagent B and reagent C.

[0049] Reagent B contains 10x PCR buffer (with MgCl 2 15mM), specific primer pair VpS and VpA, dNTP (a mixture of four deoxynucleotides), bovine serum albumin and sterile double distilled water, the contents are 5.0μl, 1.0μl (10μM), 1.0μl (10μM) respectively , 1.0 μl (10 μM each), 1.0 μl (5 mg / ml) and 38.75 μl.

[0050] Reagent C: Taq enzyme, 0.25 μl (5 units / μl).

[0051] (c) Electrophoresis and chromogenic reagents, including reagent D, reagent E and reagent F.

[0052] Reagent D: 50 times TAE (electrophoresis buffer), containing trihydroxymethylaminomethane-acetic acid and ethylenediaminetetraacetic acid,...

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 present invention provides a kit for rapid detection of Vibrio proteolyticus, which includes enrichment medium, polymerase chain reaction reagents, electrophoresis and chromogenic reagents: the present invention also provides rapid detection of Vibrio proteolyticus using this kit. The proteovibrio method has the advantages of rapidity, accuracy and high sensitivity. The method of the invention is convenient for detection, and the step of extracting bacterial DNA in the usual method is omitted; the detection sensitivity is extremely high, and the lower limit of detection is as low as 102 bacteria. The primers provided by the invention have no amplification signal for detection samples not containing Vibrio proteolyticus, and have good specificity. The invention can be applied to the detection of various samples, can detect the infection of the vibrio proteolyticus in cultured animals, and can also monitor the vibrio proteolyticus in the cultured water environment.

Description

technical field [0001] The invention relates to a kit for rapidly detecting Vibrio proteolyticus and a method for rapidly detecting Vibrio proteolyticus using the kit. Background technique [0002] Vibrio is the main pathogen of marine animals, and it has been found that Vibrio proteolyticus is one of the pathogens that cause a large number of deaths in near river oysters. Monitoring Vibrio proteolyticus in farmed animals and seawater is an important measure to prevent and control the diseases caused by it. At present, the techniques for identification of Vibrio include: 1. Classical physiological and biochemical identification methods, which are cumbersome, time-consuming, and require operators to have good professionalism; 2. Bacteria rapid automatic identification systems, which require expensive instruments and equipment and Skilled operators; 3. Nucleic acid-based detection technology, including DNA probe and PCR technology (polymerase chain reaction technology), has t...

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): C12Q1/04C12Q1/68G01N27/447G01N21/33
Inventor 张其中张占会
Owner JINAN UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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