Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

PSR detecting method for pseudomonas aeruginosa, special primers and kit

A Pseudomonas aeruginosa and kit technology, applied in the biological field, can solve problems such as the advent of PSR-specific primers and kits for Pseudomonas aeruginosa, and achieve the goal of being suitable for wide-scale popularization and application, broad market prospects, and simple operation. Effect

Inactive Publication Date: 2016-01-06
INST OF PLA FOR DISEASE CONTROL & PREVENTION
View PDF2 Cites 11 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are no PSR-specific primers and kits for detecting Pseudomonas aeruginosa on the market.

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
  • PSR detecting method for pseudomonas aeruginosa, special primers and kit
  • PSR detecting method for pseudomonas aeruginosa, special primers and kit
  • PSR detecting method for pseudomonas aeruginosa, special primers and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Primer design for PSR detection of Pseudomonas aeruginosa

[0047] Obtain the Pseudomonas aeruginosa sequence (GenBank number: NC_002516.2) from the U.S. gene database retrieval, carry out homology analysis by BLAST software, obtain the specific conserved target sequence ToxA gene of Pseudomonas aeruginosa (sequence 1 in the sequence table ), and then use the software PrimerdesignV4 to design primers for PSR detection of Pseudomonas aeruginosa according to the conserved target DNA sequence.

[0048] Table 1 is used for the primer that Pseudomonas aeruginosa is carried out PSR detection

[0049] Primer name

[0050] The above sequence 2-sequence 5 were artificially synthesized.

Embodiment 2

[0051] Embodiment 2, establishment of the PSR detection method of Pseudomonas aeruginosa of the present invention

[0052] Carry out PSR detection to Pseudomonas aeruginosa with the five primers that are used for the PSR detection of Pseudomonas aeruginosa obtained in Example 1, to obtain the best reaction system and reaction conditions, the specific methods are as follows:

[0053] 1. Determination of the optimal reaction primer concentration

[0054] Under the same reaction conditions (keep at 63°C for 60 minutes), add different concentrations of primer combinations to the reaction system to determine the optimal reaction primers and their concentrations, as follows:

[0055] 1. PSR response

[0056] Template preparation: Promega's WizardGenomicDNApurificationKit commercial DNA extraction kit was used to extract the genomic DNA of Pseudomonas aeruginosa;

[0057] PSR reaction system: with Pseudomonas aeruginosa genomic DNA as a template, PSR amplification is carried out un...

Embodiment 3

[0079] Embodiment 3, the specificity and sensitivity detection of the PSR detection method of Pseudomonas aeruginosa of the present invention

[0080] One, the specificity detection of the PSR detection method of Pseudomonas aeruginosa of the present invention

[0081] 1. PSR response

[0082] 模板制备:提取如下菌株的基因组DNA:1,P.aeruginosaATCC15442;2,P.aeruginosaCMCC10539;3,PseudomonasfluorescensCGMCC1.1802;4,Burkholderiapseudomallei029;5,PseudomonasgeniculateCGMCC1.871;6,PseudomonasmendocinaCGMCC1.593;7,Pseudomonasputida2309;8, KlebsiellapneumoniaeATCC2146;9,Streptococcuspneumoniae112-07;10,Mycobacteriumtuberculosis005;11,Staphylococcusaureus2740;12,Acinetobacterbaumannii12101;13,Escherichiacoli44825;14,Shigellaflexneri4536;15,StenotrophomonasmaltophiliaK279a;16,Legionellapneumophila9135;17,HaemophilusinfluenzaATCC49247;18,Salmonellatyphi9275;19,ProteusvulgarisCMCC49027;

[0083] PSR reaction system: the best detection system determined with embodiment 2 is the same;

[0084] The PSR re...

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 PSR detecting method for pseudomonas aeruginosa, special primers and a kit and provides polymerase spiral reaction primers for detecting pseudomonas aeruginosa, wherein the polymerase spiral reaction primers include a primer 1, a primer 2, a primer 3 and a primer 4, wherein the nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively a sequence 2, a sequence 3, a sequence 4 and a sequence 5 in a sequence table. The PSR detecting method can be used for rapidly, conveniently and efficiently detecting pseudomonas aeruginosa at high specificity and sensitivity under an isothermal condition, can be used for screening and detecting pseudomonas aeruginosa by primary medical and public health service units and various disease prevention and control centers, has wide market prospect and relatively high economic and social benefits and is suitable for wide-range popularization and application; no complex instruments are needed; and a novel technological platform is provided for detecting pseudomonas aeruginosa.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular biology detection method of bacteria, in particular to a PSR detection method of Pseudomonas aeruginosa and its special primers and kit. Background technique [0002] The scientific name of Pseudomonas aeruginosa is Pseudomonasaeruginosa, which is a common opportunistic pathogen and belongs to non-fermenting Gram-negative bacilli. The bacteria are slender and of different lengths, sometimes club-shaped or linear, arranged in pairs or short chains. There is a single flagella at one end of the bacterium, and the bacterial movement can be seen under a dark-field microscope or a phase-contrast microscope. [0003] This bacterium is an obligate aerobic bacterium, the growth temperature ranges from 25 to 42°C, and the optimum growth temperature is 25 to 30°C, especially the characteristics that the bacteria does not grow at 4°C but can grow at 42°C can be used for ...

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 Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/385
CPCC12Q1/6844C12Q2527/101
Inventor 黄留玉刘威董德荣邹大阳杨展黄思妺刘宁伟贺晓明敖大杨文超徐雅晴唐玥马文秦日辉
Owner INST OF PLA FOR DISEASE CONTROL & PREVENTION
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
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