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

Rapid qualitative and quantitative detection kit, detection method and application of bifidobacteria added in feed

A bifidobacteria, quantitative detection technology, applied in the direction of microorganism-based methods, microorganism determination/inspection, biochemical equipment and methods, etc., can solve the problem that the qualitative and quantitative detection methods are not scientific and rapid, and limit the development of probiotics preparations , consume a lot of time and resources, etc., to achieve the effect of low detection standard, high sensitivity and good accuracy

Active Publication Date: 2019-02-22
武汉泛华生物技术有限公司
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, probiotic preparations continue to develop, but the qualitative and quantitative detection methods are not scientific and fast enough, which limits the development of probiotic preparations to a certain extent.
Usually, the traditional methods of physiological and biochemical reactions and selective medium pure culture isolation and counting are very easy to limit the detection efficiency and unstable detection results due to differences in bacterial species. In addition, traditional methods will also consume a lot of time and effort. resource

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 qualitative and quantitative detection kit, detection method and application of bifidobacteria added in feed
  • Rapid qualitative and quantitative detection kit, detection method and application of bifidobacteria added in feed
  • Rapid qualitative and quantitative detection kit, detection method and application of bifidobacteria added in feed

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Selection of detection genes for bifidobacteria:

[0027] The present invention has all the probiotics such as Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus casei, Enterococcus faecium, Enterococcus faecalis, etc. A large number of comparisons and analyzes were carried out to select the target gene for qualitative and quantitative detection. The target gene is a conserved housekeeping gene in the bifidobacterium genome, and it is a single-copy gene. A specific gene was designed based on the conserved sequence of the gene. The qualitative and quantitative detection of bifidobacteria in the feed can be carried out with specific primers, and the obtained data can truly reflect the quantity of bifidobacteria added in the feed. The primers designed for the target gene are: 5'-GTCATCTGCCCGACATCTACAA-3', 5'-TAGGCTTCAGAGCAACGCAAC-3, which can be used for qualitative and quantitative detection of bifidobacte...

Embodiment 2

[0029] Rapid qualitative and quantitative detection kit for Bifidobacteria added in feed, including primers: 5'-GTCATCTGCCCGACATCTACAA-3', 5'-TAGGCTTCAGAGCAACGCAAC-3.

[0030] Example: 3:

[0031] The detection method of the rapid qualitative and quantitative detection kit for bifidobacterium added in feed, including the method of feed sample pretreatment, the method includes:

[0032] Mix 25g of the fully pulverized and homogeneous feed with 225mL of sterilized physiological saline, shake at 4°C for 1-2h at a speed of 100 rpm, and prepare a 10% uniform dilution. Aseptically perform 10-fold incremental dilution on the homogeneous dilution prepared in the previous step, select more than 2 to 3 appropriate dilutions, and extract bacterial genomic DNA or RNA as needed.

[0033] The remaining steps utilize the primers in Example 1 to carry out qualitative and quantitative identification of the extracted DNA or RNA.

[0034] The detection method for the rapid qualitative and quan...

Embodiment 4

[0095] Kit specific detection:

[0096] 1) Specificity verification of qualitative detection of bifidobacteria:

[0097]Using the primers and methods provided in Example 3, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus bulgaricus, Lactobacillus casei, Enterococcus faecium, Enterococcus faecalis, Escherichia coli, positive control (the plasmid containing the gene of interest is a template) , Bifidobacterium animalis, feed added with Bifidobacterium animalis; Negative control (sterilized water) carries out PCR amplification.

[0098] The result shows: only swimming lane 7 (feed that is added with Bifidobacterium animalis), 8 positive controls (the plasmid that contains target gene is template), 9 (Bifidobacterium animalis) has produced the amplified band of specificity 256bp, and None of the other bacterial strains showed that the primers provided by the present invention can only specifically detect bifidobacteria. At the same time, the amplification resul...

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 rapid qualitative and quantitative detection kit, detection method and application for bifidobacteria added in feed. Through comparison and analysis of reported bifidobacterium genome sequences, the invention provides a bifidobacterium Primers 5'-GTCATCTGCCCGACATCTACAA-3', 5'-TAGGCTTCAGAGCAACGCAAC-3' for the qualitative and quantitative detection of bacilli, and the invention provides a pretreatment method for feed samples, so that the probiotics in the feed can be fully released, accurate and real reactions The number of probiotics in the feed. The invention can simply, quickly and accurately qualitatively and quantitatively detect bifidobacteria in pig feed without performing pure culture of probiotics, and has simple detection procedures, high detection efficiency, good accuracy, high sensitivity and good repeatability.

Description

technical field [0001] The invention belongs to the technical field of animal husbandry and veterinary medicine. More specifically, the present invention relates to a rapid qualitative and quantitative detection kit, detection method and application for bifidobacteria added in feed. Background technique [0002] With the prohibition of antibiotics in animal feed in my country, the development of new green additives has become a hot spot. At present, probiotic preparations are constantly developing, but the qualitative and quantitative detection methods are not scientific and fast enough, which limits the development of probiotic preparations to a certain extent. Usually, the traditional methods of physiological and biochemical reactions and selective medium pure culture isolation and counting are very easy to limit the detection efficiency and unstable detection results due to differences in bacterial species. In addition, traditional methods will also consume a lot of time...

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): C12Q1/689C12Q1/04C12R1/01
CPCC12Q1/689
Inventor 吴涛侯永清赵迪丁斌鹰易丹王蕾陈洪波
Owner 武汉泛华生物技术有限公司
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