Nested PCR detection method or/and identification method of brucella

A Brucella, to-be-detected technology, applied in the field of molecular biology, can solve the problems of complex operation process, high false positive rate and false negative rate, difficult detection, etc., and achieve high sensitivity, good specificity, and simple test operation. Effect

Inactive Publication Date: 2019-02-05
ICDC CHINA CDC
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The trace amount of nucleic acid DNA in the sample to be tested is difficult to detect by ordinary PCR and multiplex PCR methods with a single primer. For pure bacterial DNA, it is difficult to identify the biotype of Brucella with a single primer. The process is complicated, and trace amounts of DNA cannot be detected; fluorescent quantitative PCR technology is a nucleic acid quantitative dete

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
  • Nested PCR detection method or/and identification method of brucella
  • Nested PCR detection method or/and identification method of brucella
  • Nested PCR detection method or/and identification method of brucella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] In the embodiment of the present invention, the nested PCR simultaneously detects and identifies Brucella species and biotypes. First, according to the transposase orfA and transposase orfB gene sequences registered in GenBank, the Primer 5.0 software is used to design the nested PCR first and second respectively. Secondary amplification primers.

[0049] Primers for the first PCR amplification:

[0050] Forward primer: F1: 5'-AACGTAACCAGATCATAGCGCATG-3', as shown in SEQ ID NO: 1;

[0051] Reverse primer: R1: 5'-ACAGATGAGCAATGGAACCGGAT-3', as shown in SEQ ID NO: 2;

[0052] Primers for the second PCR amplification:

[0053] Forward primer: F2: 5'-GAATGCGGTCAATGTTTTCTCGC-3', as shown in SEQ ID NO: 3;

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 nested PCR detection method or/and an identification method of brucella, and relates to the technical field of molecular biology. The nested PCR technique of the invention can not only detect and identify pure bacterial nucleic acid DNA, but also detect and identify trace brucella nucleic acid DNA of tissue fluid, blood, and the like; and can not only identify brucella species of the nucleic acid DNA of the sample to be tested, but also identify the biotype thereof. The nested PCR result analysis can be standardized, with simple test operation, and can be completed by a person with ordinary PCR techniques.

Description

technical field [0001] The present invention relates to the technical field of molecular biology, and the present invention relates to detection and identification Brucella bacterial strain and the nested PCR technique method of Brucella DNA in the detection sample (as: blood, serum, interstitial fluid, liver spleen etc.), by Sequence the PCR amplification products to identify the Brucella species and biotypes. The method can be used to quickly diagnose the brucellosis infection from the etiology, and identify the brucella species and biotypes. Background technique [0002] Brucellosis, referred to as brucellosis, also known as geothermal relaxation fever, Malta fever or wave fever, commonly known as "lazy man's disease", is a disease caused by intracellular parasitic bacteria of the genus Brucella. A zoonotic infection—allergic disease, brucellosis is widely distributed all over the world. [0003] More than 60 kinds of livestock, poultry, and wild animals are currently k...

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): C12Q1/6848C12Q1/04G16B40/00C12R1/01
CPCC12Q1/6848C12Q2531/113C12Q2549/119
Inventor 田国忠姜海朴东日赵鸿雁杨晓雯
Owner ICDC CHINA CDC
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