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

A rapid screening method for anemone toxin-producing microorganisms

A screening method and microbial technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of heavy workload, long cycle, and high analysis cost, and achieve high accuracy, high specificity, and screening cycle short effect

Active Publication Date: 2022-07-15
舟山出入境检验检疫局综合技术服务中心
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The key technology is the rapid screening of toxin-producing strains, which can be achieved mainly through chemical analysis of metabolites. This method not only has a large workload and a long cycle, but also the toxin standard used in chemical analysis is expensive and the analysis cost is high.
At present, there is no rapid screening technology for anemone toxin-producing strains, so it is of great significance to explore effective rapid screening methods for anemone toxin-producing microorganisms

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
  • A rapid screening method for anemone toxin-producing microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 (sensitivity test of the present invention)

[0043] (1) Sample: Marine bacteria Nioella sp.LZ7-4 (CCTCC AB 2017231) was purchased from the China Type Culture Collection, and chemical analysis of its metabolites confirmed toxin production.

[0044] (2) Sample treatment: Pick a single colony on the plate of the LZ7-4 strain, extract its genomic DNA sample according to the conventional molecular cloning method, and dilute it 8 times for later use.

[0045] (3) Obtain strain DNA: Take 1 μL and 2 μL of the above bacterial DNA samples and spot them on 6*6cm 2 The nitrocellulose filter membrane was immersed in 0.42M NaOH solution for 6 minutes, then the membrane was taken out and placed in an oven at 60°C for 1 hour to fix bacterial DNA. Then it was placed in a solution of lysozyme (5 mg / mL, pH 8.2) and incubated at 37° C. for 15 min. The bacterial cell residues on the membrane surface were washed away with TE buffer.

[0046] (4) Obtaining a DNA probe hybridizat...

Embodiment 2

[0050] Example 2 (sensitivity test of the present invention)

[0051] (1) Sample: Marine bacteria Nioella sp.LZ7-4 (CCTCC AB 2017231) was purchased from the China Type Culture Collection, and chemical analysis of its metabolites confirmed toxin production.

[0052] (2) Sample treatment: Pick a single colony on the plate of the LZ7-4 strain, extract its genomic DNA sample according to the conventional molecular cloning method, and dilute it 5 times for use.

[0053] (3) Obtain strain DNA: Take 1 μL and 2 μL of the above bacterial DNA samples and spot them on 6*6cm 2 The nitrocellulose filter membrane was immersed in 0.4M NaOH solution for 7 minutes, then the membrane was taken out and placed in an oven at 62°C for 1.1 hours to fix bacterial DNA. Then it was placed in a solution of lysozyme (5.1 mg / mL, pH 8.3) and incubated at 36°C for 14 min. The bacterial cell residues on the membrane surface were washed away with TE buffer.

[0054] (4) Obtaining the DNA probe hybridizatio...

Embodiment 3

[0058] Example 3 (sensitivity test of the present invention)

[0059] (1) Sample: Marine bacteria Nioella sp.LZ7-4 (CCTCC AB 2017231) was purchased from the China Type Culture Collection, and chemical analysis of its metabolites confirmed toxin production.

[0060] (2) Sample treatment: Pick a single colony on the plate of the LZ7-4 strain, extract its genomic DNA sample according to the conventional molecular cloning method, and dilute it 10 times for use.

[0061] (3) Obtain strain DNA: Take 1 μL and 2 μL of the above bacterial DNA samples and spot them on 6*6cm 2 The nitrocellulose filter membrane was immersed in 0.44M NaOH solution for 8 minutes, then the membrane was taken out and placed in an oven at 65°C for 1.2 hours to fix bacterial DNA. Then it was placed in a solution of lysozyme (5.2 mg / mL, pH 8.4), and incubated at 38° C. for 18 min. The bacterial cell residues on the membrane surface were washed away with TE buffer.

[0062] (4) Obtaining a DNA probe hybridiza...

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 relates to the field of marine biotechnology, and discloses a rapid screening method for anemone toxin-producing microorganisms in view of the technical gap of a rapid screening method for anemone toxin-producing microorganisms, comprising the following steps: (1) obtaining strain DNA; ( 2) Obtain DNA probe hybridization solution: put the digoxigenin-labeled DNA probe solution in boiling water and then put it in an ice bath, take the DNA probe solution and add it to the hybridization solution; (3) Hybridization: place the DNA membrane Place in DNA probe hybridization solution and incubate for hybridization; place the hybridized DNA membrane in a container containing 2 x SSC solution for washing; (4) develop color; (5) judge the result. The DNA probe-colony in situ hybridization method is used for the rapid screening of sea anemone toxin-producing microbial strains, and the strains that can produce sea anemone toxin can be accurately screened through specific amplified DNA probes and simple and orderly operation steps , this method is simple and efficient, with short screening period and high accuracy.

Description

technical field [0001] The invention relates to the technical field of marine biology, in particular to a rapid screening method for anemone toxin-producing microorganisms. Background technique [0002] Palytoxins (PLTXs) are one of the most toxic non-protein algal toxins known, and are listed as one of the largest non-polymeric natural products; they are highly toxic, fat-soluble, heat-stable polyether marine biotoxins ; is one of the most widely distributed and highest incidence marine toxins. It can cause irreversible damage to various organs and tissues such as the intestines, liver, nerves, placenta, etc., and there is no specific drug treatment after poisoning. Sea anemone toxins seriously endanger the safety of marine ecosystems and human health, and have become a worldwide research hotspot. Sea anemone toxin is a neurotoxin and has important applications in the detection of harmful red tide, neurophysiology, medical diagnosis, drug development, and biochemical warfa...

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): C12Q1/6841C12Q1/04
CPCC12Q1/6841C12Q2527/125C12Q2563/131C12Q2565/518
Inventor 张静周秀锦邵宏宏胡兴娟
Owner 舟山出入境检验检疫局综合技术服务中心
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