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

Genetic diagnostic kit and detection method of Pasteurella multocida, a major bovine pathogen

A diagnostic kit, Pasteurella technology, applied in the field of biological sciences, can solve the problems of troublesome preparation, limited application and development, long detection time, etc., to achieve the effect of ensuring rapidity, high practical value, and avoiding the spread of germs

Active Publication Date: 2017-08-15
KUNMING DIANLONG BIOLOGICAL MEDICINE SCI & TECH
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some of these detection methods have high technical requirements and long detection time.
The preparation of some required materials is troublesome and expensive, and some methods have low sensitivity and specificity. Therefore, the techniques mastered by veterinarians and cattle breeding owners are very difficult and difficult to promote, which limits the application and development of these techniques.

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
  • Genetic diagnostic kit and detection method of Pasteurella multocida, a major bovine pathogen
  • Genetic diagnostic kit and detection method of Pasteurella multocida, a major bovine pathogen
  • Genetic diagnostic kit and detection method of Pasteurella multocida, a major bovine pathogen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A genetic diagnostic kit for a serious bovine pathogen-Pasteurella multocida, which includes liquid A, liquid B, liquid C, a box and a foam board;

[0036] Said liquid A is a sample diluent, filled with sterile physiological saline, 5 tubes, 5ml / tube;

[0037] The B solution is the PCR reaction solution, 1 tube, containing 192µl PCR amplification reaction solution, including 168µl ddH 2 O, 14.4 µl with Mg2+ 10×Buffer [Tris-HCl 100mmol / L, pH 9.0, KCl 500mmol / L, MgCl 2 15mmol / L,

[0038] Triton-X100 1%], 3.2 µl of dNTP at a concentration of 250 μmol / L, 2.4 µl of a primer PmF at a concentration of 0.5 μmol / L, 2.4 µl of a primer PmR at a concentration of 0.5 μmol / L, and 1.6 µl of TaqE with an enzyme activity unit of 8 U; It is: 5'-ACGCTTTGCCTT GCAAGATAC-3', PmR is: 5'-GATTCACGTAACAAACCAGGC-3'; the C solution is a positive control solution, 1 tube, containing 50µl Pasteurella multocida positive template;

[0039] The C solution is a positive control solution, 1 tube cont...

Embodiment 2

[0053] A method for detecting a major bovine pathogen-Pasteurella multocida, using the kit of Example 1, carried out according to the following steps:

[0054] Step (1), take 0.05g of the lung tissue of the acutely ill dead cattle of the test specimen, add the sample diluent, dilute 10 times, and homogenize in an ice bath in a sterile homogenizer;

[0055] Step (2), centrifuge the homogenized sample at a speed of 3000r / min for 5min to obtain the supernatant of the centrifuged sample;

[0056] Step (3), boil the supernatant of 200 μl centrifuged sample for 10 minutes, then immediately place it on ice for 5 minutes to cool down;

[0057] Step (4), centrifuge the cooled sample at a speed of 3000r / min for 5min, and take the supernatant as a PCR template;

[0058] In step (5), take the PCR template and C solution with a volume ratio of 1:1, respectively, and add them to the PCR reaction solution. The volume ratio of the PCR template to the PCR reaction solution is 1:24, and the vo...

Embodiment 3

[0062] A method for detecting a major bovine pathogen-Pasteurella multocida, using the kit of Example 1, carried out according to the following steps:

[0063] Step (1), take 0.05g of fresh nasal swab fluid of the tested specimen, add sample diluent, dilute 20 times, and homogenize in an ice bath in a sterile homogenizer;

[0064] Step (2), centrifuge the homogenized sample at a speed of 4000r / min for 10min to obtain the supernatant of the centrifuged sample;

[0065] Step (3), boil the supernatant of 200µl centrifuged sample for 10min, then immediately place it on ice for 10min to cool;

[0066] Step (4), centrifuge the cooled sample at a speed of 4000r / min for 10min, and take the supernatant as a PCR template;

[0067] In step (5), take the PCR template and C solution with a volume ratio of 1:1, respectively, and add them to the PCR reaction solution. The volume ratio of the PCR template to the PCR reaction solution is 1:24, and the volume of the C solution and the PCR reac...

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 provides a gene diagnosis kit and detection method for major cattle pathogenic bacteria-Pasteurella multocida and belongs to the technical field of bioscience. The kit and the detection method are designed by taking a pair of primers as the main body, and the primers are designed according to the Pasteurella multocida gene conservative area sequence. According to the invention, the polymerase chain reaction (PCR) technology is adopted for qualitatively detecting the specificity DNA fragments of cattle pathogenic bacteria-Pasteurella multocida simply, conveniently and quickly, the specificity is good and the sensitivity is high; the kit and the detection method can be used for clinical detection of cattle pasteurellosis, bacteria tracking and detecting in all cattle farming periods and monitoring of the cattle farming environment and avoid pathogen transmission and outbreak, thereby achieving a very high utility value.

Description

technical field [0001] The invention belongs to the technical field of biological sciences, and in particular relates to a diagnostic kit and a detection method for major bovine pathogenic bacteria, in particular to a genetic diagnosis kit and a detection method for Pasteurella multocida (Pasteurella multocida). Background technique [0002] Pasteurellosis of cattle, also known as bovine hemorrhagic septicemia, is an acute, febrile infectious disease of cattle, characterized by high fever, pneumonia and extensive hemorrhage of internal organs. The pathogen is Pasteurella multocida, the bacteria are small, club-shaped or short rod-shaped bacteria with blunt ends, the size is 1-1.5μm×0.3-0.6μm, mostly scattered, non-motile, and do not form spores , negative Gram stain. Pasteurella multocida is pathogenic to various livestock and wild animals, and can occur regardless of age. Cattle are susceptible to the disease with cattle and dairy cows, among which young and middle-aged d...

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): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/686C12Q2531/113
Inventor 邓先余林连兵夏雪山
Owner KUNMING DIANLONG BIOLOGICAL MEDICINE SCI & TECH
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