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

Cross primer amplification primer group for detecting haemophilus parasuis, kit and application

A cross-primer amplification, Haemophilus suis technology, applied in the biological field, can solve the problems of insufficient sensitivity, hindering the application and popularization of the technology, expensive, etc., and achieves a reaction result that is easy, easy to popularize and apply on a large scale, and low in use cost. Effect

Active Publication Date: 2017-06-13
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
View PDF7 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Angen et al. (Angen, O. and S. Oliveira, et al. (2007). "Development of an improved species specific PCR test for detection of Haemophilus parasuis." Vet Microbiol 119(2-4):266-76.) in 2006 Although the Haemophilus parasuis PCR detection technology established in 2010 can quickly detect Haemophilus parasuis, the sensitivity of this method is relatively insufficient
In addition, PCR detection technology requires expensive thermal cyclers and skilled operators during the amplification process, which hinders the wide application and promotion of this technology in veterinary grassroots

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
  • Cross primer amplification primer group for detecting haemophilus parasuis, kit and application
  • Cross primer amplification primer group for detecting haemophilus parasuis, kit and application
  • Cross primer amplification primer group for detecting haemophilus parasuis, kit and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Design and synthesis of embodiment 1 cross primer constant temperature amplification primer

[0052] 1. Primer design

[0053] According to the principle of primer design, aiming at the conserved region sequence of the wzs gene of Haemophilus parasuis (while ensuring that the conserved sequence is quite different from the nucleotide sequence of the wzs gene of Pasteurella), primers were designed using Priemer5 software, according to our experience Ensure that the GC content of the primers is between 40% and 60%, and the Tm value of each primer is around 55°C. Then we used the PrimerSelect tool of Larsergene7.0 biological software to preliminarily screen the primers. The screening principle was to ensure that the dG value between each primer pair was small. Through preliminary screening, we obtained a set of primers with better theoretical values, as shown in Table 1 (at this time, F1 was not labeled with Biotin). The primers were synthesized by Shanghai Sangon Bioengin...

Embodiment 2

[0054] Application of Example 2 Cross Primer Constant Temperature Amplification Primer in Detecting the bacterium to be tested

[0055] 1. Extraction of Haemophilus parasuis Genomic DNA by Boiling

[0056] (1) Get the serum Haemophilus parasuis type 5 preserved by freeze-drying, PBS (pH value is 7.4, concentration is 0.15M phosphate buffer saline: KH 2 PO 4 0.2g, Na 2 HPO 4 12H 2 O 2.9 g, NaCl 8 g, KCl 0.2 g) were dissolved, 50 μL was inoculated into 5 mL TSB medium, and cultured with shaking at 37 °C for 24 h.

[0057] (2) Take one loop of the inoculation loop, streak it on the TSA agar medium plate, and incubate at 37°C for 24 hours.

[0058] (3) A single colony was picked, inoculated in 5 mL TSB liquid medium, and cultured with shaking at 37°C for 18 hours.

[0059] (4) Take 1 mL of the bacterial culture and centrifuge at 4000 r / min for 5 min to collect the bacteria.

[0060] (5) Wash 3 times with 500 μL sterilized ultrapure water.

[0061] (6) Resuspend the pellet...

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 cross primer amplification primer group for detecting haemophilus parasuis, a kit and application. The primer group comprises a primer group with nucleotide sequences as shown in SEQ ID NO.1 to 6; the kit comprises the primer group and a nucleic acid test strip; an application method of the kit comprises the following steps of firstly preparing a cross primer amplification reaction system; directly reading after using the nucleic acid test strip for detecting a product obtained through thermostatic reaction, wherein a positive result is characterized by appearing two strips, one is located in a detection area, and the other one is located in a quality control area. The kit is simple to operate, low in cost, easy to observe the reaction result, good in specificity, applicable for on-site test of export quarantine, food sanitation and an animal husbandry farm, and easy to popularize and apply in large scale.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a rapid detection kit for cross-primer amplification of Haemophilus parasuis and its application. Background technique [0002] Haemophilus parasuis (Hps) is a common opportunistic pathogen in the upper respiratory tract of pigs, which can invade the body under certain conditions and cause serous, fibrinous polyserositis, arthritis, An infectious disease characterized by dyspnea, high fever and high mortality, especially for early weaned piglets. With the development of large-scale pig farming, the harm range of Haemophilus parasuis to pig farming is also expanding, which has brought huge economic losses to pig farming. [0003] Although Haemophilus parasuis disease is generally controlled through vaccination and antibiotic treatment, accurate pathogenic diagnosis is an important prerequisite for the control and purification of the disease. At present, bacterial isolati...

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): C12Q1/68C12Q1/04C12N15/11C12R1/21
CPCC12Q1/6844C12Q1/689C12Q2527/101C12Q2531/119C12Q2565/625
Inventor 勾红潮李春玲宋帅李淼蔡汝健蒋智勇李艳杨冬霞楚品品
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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