Gene chip for detecting nine pathogenicity vibrios in marine products

A pathogenic vibrio and gene chip technology, which is applied in the detection/inspection of microorganisms, resistance to vector-borne diseases, DNA/RNA fragments, etc., can solve problems such as complex operations, heavy manual labor, and long inspection cycles. Achieve the effect of strong repeatability, easy operation and high accuracy

Inactive Publication Date: 2014-05-28
ZHOUSHAN INST OF CALIBRATION & TESTING FOR QUALITY & TECHNICAL SUPERVISION
View PDF5 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection and identification methods of pathogenic bacteria in seafood in my country still remain at the routine microbiological inspection level, usually based on isolation culture, biochemical tests and serological tests, which have complex operations, heavy manual labor, and long inspection cycles (6-7 days). Days), low specificity and other shortcomings, to a considerable extent limit the ability of rapid detection and identification of food-borne disease pathogeni

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
  • Gene chip for detecting nine pathogenicity vibrios in marine products
  • Gene chip for detecting nine pathogenicity vibrios in marine products
  • Gene chip for detecting nine pathogenicity vibrios in marine products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Design and preparation of probes

[0025] 1. Sequence acquisition:

[0026] According to the species ID number of the target species, all the sequence information of the species was searched in the Genbank database, and the genbank format information of all the retrieved sequences was downloaded; the collected related genes are shown in Table 1 below.

[0027]

[0028] 2. Probe design method

[0029] 1) Search the NCBI nucleic acid database to find the gene information of all related species, and download the data in genbank format.

[0030] 2) Extract the gyrb gene sequence from the downloaded data in genbank format.

[0031] 3) For each species of microorganism in the project, use Muscle to perform multiple sequence comparisons of all its gyrb gene sequences.

[0032] 4) According to the multiple sequence alignment results of Muscle, combine the tools to obtain the consensus sequence.

[0033] 5) Take the consensus sequence as input, combine the tool...

Embodiment 2

[0050] Embodiment 2: Design and preparation of strain amplification primers

[0051] of the gene chip of the present invention

[0052] 1. Sequence acquisition: the same as the sequence of the designed probe.

[0053] 2. Primer Design

[0054](1) Design of primers for amplifying interval sequences: After comparing the 16s rRNA of nine Vibrio species downloaded from the public database NCBI with the sequence alignment software Glustal X, a 16s rRNA conserved sequence close to the interval was selected as the upstream primer. And contains Vibrio universal probe sequence.

[0055] (2) Design of primers for amplifying specific gene sequences: compare the specific gene sequences of the above nine species of Vibrio downloaded from the GenBank public database with the sequence comparison software Glustal X, find the conserved segment of the gene, and insert The conserved segment was imported into the primer design software Primer Premier5.0 software. From the output results, sele...

Embodiment 3

[0063] Example 3: Gene Chip Preparation - Chip Spotting

[0064] 1. Dissolve the probes: Dissolve each probe to 40 μM with sterile water, and mix it with the gene chip sample solution at a ratio of 1:1, so that the final concentration of each probe is 20 μM.

[0065] 2. Add plate: Add the dissolved probe to the corresponding position of the 384-well plate, 10 μL per well.

[0066] 3. Spotting: put a clean aldehydized glass slide (Boao Biotechnology) of 57.5mm×25.5mm×1mm (length×width×height) on the stage of the chip spotter (spotarray72), use the control The software (PersonalArrayer16) runs the program, spots in the spotting area on the aldehyded glass slide to form a medium-low density DNA microarray, and the array arrangement rules in the four dot matrix areas on the glass slide are the same.

[0067] 4. Drying: Dry the ordered chips overnight at room temperature, and then dry them in a 45 oven for 2 hours.

[0068] 5. Cross-linking: use a cross-linker to cross-link twice...

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 a gene chip for detecting nine pathogenicity vibrios in marine products and a kit thereof. The gene chip comprises a solid phase carrier and an oligonucleotide probe, wherein the oligonucleotide probe includes one or more selected from the following nucleotide sequences: 1) DNA sequences selected from genes of a vibrio hollisae, a vibrio vulnificus, a vibrio cholera, a vibrio parahemolyticus, a vibrio harveyi, a vibrio alginolyticus, a vibrio furnissi, a vibrio mimicus and a vibrio damsel, 2) complementary DNA sequences of the DNA sequences selected in the DNA sequences in 1), 3) complementary RNA sequences of the selected DNA sequences in 1) or 2). The kit comprises the gene chips. The gene chip for detecting nine pathogenicity vibrios in marine products and the kit thereof provided by the invention are used for detecting the nine pathogenicity vibrios in marine products, and have the advantages of simplicity in operation, good sensitivity and strong repeatability.

Description

technical field [0001] The invention belongs to the technical field of microbial molecular detection, and in particular relates to a gene chip for detecting nine kinds of pathogenic Vibrio in seafood. Background technique [0002] Zhejiang Province is a large ocean province with a rich variety of seafood. Seafood products are deeply loved by local people, and many residents have the habit of eating raw seafood. However, with the increasing pollution of the marine environment, pathogenic microorganisms are enriched in large quantities in aquatic products, and food-borne diseases and food poisoning caused by eating marine products have occurred frequently, posing a great threat to people's life and health. . Pathogenic Vibrio is the most important pathogenic microorganism in seafood, including Vibrio cholerae, Vibrio mimicus, parahaemolyticus, Hollisia, Vibrio alginolyticus, Furnaceium, Vibrio vulnificus, mermaid There are nine kinds of Vibrio and Vibrio harveylius. The path...

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
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/689C12Q2600/178Y02A50/30
Inventor 薛超波黄朱梁王萍亚
Owner ZHOUSHAN INST OF CALIBRATION & TESTING FOR QUALITY & TECHNICAL SUPERVISION
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap