SSCP (single strand conformation polymorphism) detecting method for animal liver bacterial toxin contamination

A bacterial toxin and animal liver technology, applied in the biological field, can solve the problems of high detection cost, time-consuming and laborious, and inaccurate detection results, and achieve the effects of good repeatability, low detection cost, and easy promotion and application

Inactive Publication Date: 2014-08-20
YUNNAN AGRICULTURAL UNIVERSITY
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two methods are time-consuming and laborious, the detection cost is high, and the detection results are inaccurate for different types of animals. Therefo...

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
  • SSCP (single strand conformation polymorphism) detecting method for animal liver bacterial toxin contamination
  • SSCP (single strand conformation polymorphism) detecting method for animal liver bacterial toxin contamination
  • SSCP (single strand conformation polymorphism) detecting method for animal liver bacterial toxin contamination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: The SSCP detection method of animal liver bacterial toxin contamination, the method comprises liver mitochondrial DNA extraction; in liver mitochondrial DNA, cytochrome tRNA Cys-Tyr The gene fragment at the position 5109-5299 of the gene is amplified; and the point mutation of the gene fragment is analyzed by SSCP to evaluate whether the animal liver is polluted by the bacterial toxin. Three steps. The specific implementation process of the three steps is as follows:

[0036] Step 1, liver mitochondrial DNA extraction:

[0037] First, prepare the reagents in the mitochondrial DNA extraction step, the recipe is as follows:

[0038] (1) SE solution: pH7.8, mixed with 0.25M sucrose, 2.5mM CaCl2, 30mM Tris-HCl and 10mM EDTANa2;

[0039] (2) Solution Ⅰ (TEN), pH 8.0, mixed with 0.15M NaCl, 10mM EDTANa2 and 10mM Tris-HCl;

[0040] (3) Solution II, 1% SDS, containing 0.2N NaOH;

[0041] (4) Solution Ⅲ, 3M Kac, which contains 3M KAc, 5M glacial acetic acid, mix w...

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 an SSCP (single strand conformation polymorphism) detecting method for animal liver bacterial toxin contamination, relates to the biological technical field and particularly relates to a detecting method for animal liver bacterial toxin contamination. According to the method, an SSCP technology is utilized to detect gene mutation of animal liver mitochondria contaminated by bacterial toxin, so that the technology is applied to evaluating and identifying animal food safety. Main steps comprise extracting host liver mitochondria DNA (deoxyribonucleic acid), amplifying the liver mitochondria tRNACys-Tyr gene segment by applying PCR(polymerase chain reaction)-SSCP technology, detecting a stripe with abnormal swimming motility by polyacrylamide gel electrophoresis, sequencing after purifying the stripe, and indicating that the animal food is contaminated by the bacterial toxin if the tRNACys -Tyr gene segment is inserted with insertion mutation of a basic group T at the 5238th and 5239th basic groups. The SSCP detecting method disclosed by the invention can be used for solving the defects of the conventional detecting method, and is quick, accurate, specific, sensitive, good in repeatability, low in detecting cost and easy for popularization and application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection method for animal liver bacterial toxin contamination. Background technique [0002] Bacterial infection and the abuse of antibiotics in the process of large-scale feeding of food animals often lead to bacterial infection and bacterial toxin pollution in animals. The liver is the main detoxification organ of animals and is easily contaminated by bacterial toxins. Chinese people have the habit of eating animal offal. Therefore, it is particularly necessary to detect whether animal liver is contaminated by bacterial toxins for food safety evaluation. The traditional detection method is to separate and prepare the toxin, administer it to mice, and judge it by clinical symptoms, or collect blood, and use expensive Limulus test and high-performance liquid chromatography to detect the toxin. The above two methods are time-consuming, labor-intensive, costly, and the test result...

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): C12Q1/68
CPCC12Q1/6858C12Q1/6869
Inventor 严玉霖高洪赵汝陈利平陈玲
Owner YUNNAN AGRICULTURAL UNIVERSITY
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