CN109468394A - A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups - Google Patents
A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups Download PDFInfo
- Publication number
- CN109468394A CN109468394A CN201811462307.8A CN201811462307A CN109468394A CN 109468394 A CN109468394 A CN 109468394A CN 201811462307 A CN201811462307 A CN 201811462307A CN 109468394 A CN109468394 A CN 109468394A
- Authority
- CN
- China
- Prior art keywords
- salmonella
- pcr
- seq
- sequence information
- choleraesuls
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/16—Primer sets for multiplex assays
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to pathogenic bacteria detection technique fields, disclose a kind of five heavy PCR primers, kit and its application for detecting four kinds of Salmonella serogroups.Primer includes five pairs of specific primers, it is characterised in that: five pairs of specific primers are respectively as follows: salmonella-F；Salmonella-R；Bacterium enteritidis-F；Bacterium enteritidis-R；Salmonella typhimurium-F；Salmonella typhimurium-R；Salmonella typhi-F；Salmonella typhi-R；Salmonella choleraesuls-F；Salmonella choleraesuls-R.The present invention can not only detect whether as salmonella, and just simultaneously accurately can identify whether be salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls by PCR detection merely, without other ancillary technique means.
Description
Technical field
The present invention relates to pathogenic bacteria detection technique field more particularly to a kind of five weights for detecting four kinds of Salmonella serogroups
PCR primer, kit and its application.
Background technique
Salmonella (Salmonella) is the most common food-borne pathogens, and the food poisoning case being induced by it is alive
Often occupy the first in the food poisoning of boundary various regions.In the U.S., about 1,400,000 people infect detection of Salmonella every year, and lead to 600
People is dead.Bacterium distribution is very wide, usually resides in the enteron aisle of human body and animal body, and with their existence ring of manure contamination
Border and food, to be propagated between human body and animal body.The separated serotype out in the whole world has more than 2600 kinds at present.Its
In cause human body poison by food Salmonella serogroup bacterial strain focus primarily upon A~D group, account for Foodborne salmonella
70%, it is especially most commonly seen with salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls etc..
Currently, the classifying method of salmonella can be divided into two class of Phenotype typing and molecule parting.Phenotype typing method is main
Including serotype and bacteriophage typing.Since first time in 1934 delivers Kauffmann-White serum table, Salmonella
The serotype of bacterium just becomes most important parting form.The serotype of salmonella is the whip based on its somatic cells surface
Hair, pod membrane or rete malpighii, the albumen of purifying etc. generate different congealing reactions from specific corrosioning anteserum and distinguish.Serum slide is solidifying
Collection test is used as early stage salmonella classifying method, intuitive and reliable, but there is also shortcomings: (1) qualification time is long: determining one
The serotype of strain salmonella needs 1-2 days, and if desired some bacterial strains make flagellum induction then needs one week or so.(2) bacterial strain itself:
Certain bacterial strains morph, and become rough type from smooth type, are produced from coagulation phenomena, can not parting.(3) there are the mistakes such as cross agglutination
Accidentally result.(4) experimenter's operating experience:, reaction interval more slow to the agglutinating reaction of some Salmonella antigens and serum
Spend situations such as weaker, it is necessary to which veteran experimenter identifies, otherwise easily causes the genotyping result of mistake.(5) city
Commercialization salmonella diagnostic serum on field is expensive, and quality is irregular, and included serotype is uneven, and the source of goods is tight
It lacks.
Salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls are weights in Salmonella
The highly pathogenic bacterium wanted.But the similarity degree of both Salmonella choleraesuis and salmonella paratyphi C genome reaches
95% or more.According to the PCR detection method of the Salmonella choleraesuls retrieved at present, it all cannot distinguish between Salmonella choleraesuls
And moscow' paratyphi C, in the prior art, if to further discriminate between Salmonella choleraesuls and paratyphoid C
Salmonella, it is necessary to add supplementary means again on the basis of PCR.What such as Lin Yiman was delivered " real-time fluorescence PCR while detecting
The foundation and application of salmonella paratyphi C and Salmonella choleraesuis method " in, " one real-time fluorescence PCR of amelioration regionalization
Salmonella choleraesuis and salmonella paratyphi C are detected simultaneously " in, it is on the basis of PCR detection again by real-time fluorescence
Technology could effectively distinguish Salmonella choleraesuls and moscow' paratyphi C.
Summary of the invention
In order to solve the above-mentioned technical problems, the present invention provides a kind of five heavy PCR for detecting four kinds of Salmonella serogroups
Primer, kit and its application, the present invention can not only detect whether as salmonella, and can pass through merely PCR and detect
Just simultaneously accurately identify whether be salmonella typhimurium, Bacterium enteritidis, salmonella typhi, hog cholera Salmonella
Bacterium, without other ancillary technique means.
The specific technical proposal of the invention is: a kind of five heavy PCR primers for detecting four kinds of Salmonella serogroups, including five
To specific primer, it is characterised in that: five pairs of specific primers are respectively as follows:
Salmonella-F, sequence information is as shown in SEQ ID No.1；
Salmonella-R, sequence information is as shown in SEQ ID No.2；
Bacterium enteritidis-F, sequence information is as shown in SEQ ID No.3；
Bacterium enteritidis-R, sequence information is as shown in SEQ ID No.4；
Salmonella typhimurium-F, sequence information is as shown in SEQ ID No.5；
Salmonella typhimurium-R, sequence information is as shown in SEQ ID No.6；
Salmonella typhi-F, sequence information is as shown in SEQ ID No.7；
Salmonella typhi-R, sequence information is as shown in SEQ ID No.8；
Salmonella choleraesuls-F, sequence information is as shown in SEQ ID No.9；
Salmonella choleraesuls-R, sequence information is as shown in SEQ ID No.10.
As described in the application background technique, both Salmonella choleraesuis and salmonella paratyphi C genome it is similar
Degree is up to 95% or more.According to the PCR detection method of the Salmonella choleraesuls retrieved at present, hog cholera sand all cannot distinguish between
Door Salmonella and moscow' paratyphi C, in the prior art, if to further discriminate between Salmonella choleraesuls and the third type
Salmonella paratyphi, it is necessary to add supplementary means again on the basis of PCR.Such as Lin Yiman deliver " real-time fluorescence PCR is same
When detect salmonella paratyphi C and Salmonella choleraesuis method foundation and application " in, " amelioration regionalization one in real time it is glimmering
Light PCR detects Salmonella choleraesuis and salmonella paratyphi C simultaneously " in, it is on the basis of PCR detection again by reality
When fluorescent technique, could effectively distinguish Salmonella choleraesuls and moscow' paratyphi C.
And in the present invention, team of the present invention is by a large amount of theoretical research and test, to the sequence of specific primer
Design optimizes, it was found that the positive anti-primer of above-mentioned Salmonella choleraesuls can effectively distinguish Salmonella choleraesuls and third
Type salmonella paratyphi, without other technologies means.The final resulting five pairs of primers of the present invention, are interfered between each other
Small, non-specific amplification reacts less, sensitivity and specificity are high.Inventors believe that five pairs of specific primer solutions of the invention
It has determined not relying on PCR merely in the prior art and detect and Salmonella choleraesuls and moscow' paratyphi C cannot be distinguished
Technical problem achieves significant progress.
A kind of kit using above-mentioned five weights PCR primer, including five heavy PCR reaction systems, the five heavy PCR reactant
System is contained in terms of 50 μ L of total volume:
2 × Multiplex PCR Mix1,25 μ L,
0.25 μ L of Multiplex PCR Mix2,
- F0.4 μM of salmonella,
- R0.4 μM of salmonella,
- F0.4 μM of Bacterium enteritidis,
- R0.4 μM of Bacterium enteritidis,
- F0.4 μM of salmonella typhimurium,
- R0.4 μM of salmonella typhimurium,
- F0.4 μM of salmonella typhi,
- R0.4 μM of salmonella typhi,
- F0.4 μM of Salmonella choleraesuls,
- R0.4 μM of Salmonella choleraesuls,
4 μ L of template,
Moisturizing is to 50 μ L.
A kind of application detected in four kinds of Salmonella serogroups using mentioned reagent box in PCR.
Preferably, PCR amplification condition are as follows: 94 DEG C of 1min；94℃30sec；57℃ 30sec；72℃ 45sec；35
Circulation, 72 DEG C of 10min；4℃；1 circulation.
On the basis of obtaining five pairs of specific primers, then cooperate the optimization in mentioned reagent box and PCR amplification condition,
Sensitivity and accuracy can be further increased.
It is compared with the prior art, the beneficial effects of the present invention are: the present invention can not only detect whether as salmonella, and
And just simultaneously accurately can identify whether be salmonella typhimurium, Bacterium enteritidis, typhoid fever sand by PCR detection merely
Door Salmonella, Salmonella choleraesuls, without other ancillary technique means.
Detailed description of the invention
Fig. 1 is that five weight PCR detect 4 kinds of different serotypes salmonella results；
Fig. 2 is that five weight PCR detect 10 kinds of different serotypes salmonella results；
Fig. 3 is the specificity result that five weight PCR detect other bacteriums.
Specific embodiment
The present invention will be further described with reference to the examples below.
Embodiment
1 materials and methods
Pathogen is used in 1.1 tests
Salmonella typhi (CICC10867), moscow' paratyphi C (CICC21512), salmonella typhimurium
(CICC21483), Bacterium enteritidis (CICC24119), Salmonella choleraesuls (CICC21493), vibrio parahemolyticus
(ATCC17802), staphylococcus aureus (ATCC6538), Listeria monocytogenes (ATCC19115), Vibrio vulnificus
(VV001), Shigella flexneri (ATCC12022), Escherichia coli (ATCC25922), vibrio mimicus (VM001), sonnei will are congratulated
Bacterium (ATCC51592), staphylococcus epidermis, enterococcus (ATCC33186), citrobacter freundii irrigate formula staphylococcus, pneumonia
Klebsiella.
1.2 key instruments and reagent
(Dalian is precious by multiplexed PCR amplification kit RR062A (the limited work department of Dalian treasured bioengineering), DL1000bp DNAMarker
The limited work department of bioengineering), agarose (the limited work department of Dalian treasured bioengineering), PCR amplification instrument, gel imager, electrophoresis apparatus
And mating electrophoresis tank, supercentrifuge.
1.3 design of primers
It is upper from the gene pool of NCBI to download salmonella typhimurium, Bacterium enteritidis, salmonella typhi, hog cholera sramana
The target-gene sequence of Salmonella makees the conserved regions design specific primer after sequence analysis in each salmonella target gene.Primer
Synthesis commission Dalian treasured Biotechnology Co., Ltd completes, and DPO primer sequence is shown in Table 1.
The DPO primer sequence of the weight of table 1 five PCR
1.4 pathogens quantify the preparation of bacterium solution
By the bacterium reference culture culture of above-mentioned cause of disease to logarithmic phase, the every mL clump count surveyed by LB agar plate count, by bacterium
Sample carries out 10 times of serial dilutions to 10 with physiological saline respectively1CFU/mL。
1.5 pathogenic bacteria gene group DNA are extracted
Above-mentioned Quantifying Bacteria suspension 1mL is taken to be put into 1.5mL eppendof pipe, 15000rpm high speed centrifugation 1min goes to reset and add
After entering 100 μ L of DNA extractant mixing, 2min in boiling water, 15000rpm high speed centrifugation 2min, supernatant, that is, DNA solution (PCR are set
Template).
The heavy PCR reaction system optimization of 1.6 primer five and foundation
2 × Multiplex PCR Mix1,25 μ L, Multiplex PCR Mix2,0.25 μ L, 5 pairs of primer final concentrations after optimization
It is respectively as follows: 0.4 μM of salmonella-F, 0.4 μM of salmonella-R, 0.4 μM of Bacterium enteritidis-F, Bacterium enteritidis-R
0.4 μM, 0.4 μM of salmonella typhimurium-F, 0.4 μM of salmonella typhimurium-R, 0.4 μM of salmonella typhi-F, typhoid fever
0.4 μM of salmonella-R, 0.4 μM of Salmonella choleraesuls-F, 0.4 μM of Salmonella choleraesuls-R, 4 μ L of template, moisturizing is extremely
50 μ L, are expanded in PCR instrument after centrifugation；Condition are as follows: 94 DEG C of 1min, 94 DEG C of 30sec；57℃ 30sec；72℃
45sec；35 circulations, 72 DEG C of 10min；4℃；1 circulation.End of reaction, the observation of 2% agarose gel electrophoresis is as a result, special
Property band judges reference standard molecular weight (DL1000).
The specificity of 1.7 primer PCRs
1.7.1 the specificity inside five weight PCR
The DNA for expanding 5 kinds of target genes one by one with five weight PCR reaction systems has seen whether that non-specific amplification band generates and has come
Specificity inside five weight PCR of verifying.
1.7.2 it is anti-to carry out five weight PCR using non-targeted salmonella for the specificity that five weight PCR detect other salmonellas
It answers, has seen whether non-specific amplification band.
Table 2-2 is for trying different serotypes salmonella
1.7.3 five weight PCR detect the specificity of other bacteriums
With five heavy PCR reaction systems respectively to vibrio parahemolyticus, staphylococcus aureus, Listeria monocytogenes, Vibrio vulnificus,
Shigella flexneri, Escherichia coli, vibrio mimicus, sonnei shigella dysenteriae, staphylococcus epidermis, enterococcus, citrobacter freundii,
The genomic DNA of Klebsiella Pneumoniae makees five weight PCR reactions, has seen whether non-specific amplification band.
2 results
2.1 the method that five weight PCR detect 4 kinds of pathogenic bacteria simultaneously is established
The heavy PCR reaction system of optimized five can simultaneously to salmonella typhimurium, Bacterium enteritidis, salmonella typhi,
Salmonella choleraesuls are detected, and realize a Guan Duojian.The specific band and expected amplification length one that five weight PCR are expanded
Cause (Bacterium enteritidis 221bp, Salmonella choleraesuls 342bp, salmonella typhi 532bp, salmonella typhimurium
656bp, salmonella species-specific genes 856bp, is shown in Fig. 1, wherein M:DL1000,1 Bacterium enteritidis, 2 hog cholera sramana
3 salmonella typhi of Salmonella, 4 salmonella typhimurium, 5 Bacterium enteritidis, Salmonella choleraesuls, salmonella typhi, mouse
Salmonella typhi mixing.
The specificity of 2.2 5 weight PCR detection methods
2.2.1 the specificity inside five weight PCR
The five weight PCR methods established can effectively expand Bacterium enteritidis, Salmonella choleraesuls, salmonella typhi, mouse wound
Cold salmonella obtains five specific amplification bands, compares target base when being in design after above-mentioned band sequencing through BLAST
Because in sequence context.See Fig. 1.
2.2.2 five weight PCR detect the specificity of other salmonellas
Using non-targeted salmonella, five weight PCR reactions are carried out, have seen whether non-specific amplification band.See Fig. 2, wherein
M:DL1000；1: moscow' paratyphi C；2:ZJZSJK-201708；3:ZJZSJK-201709；4:ZJZSJK-
201711；5:ZJZSJK-201713；6:ZJZSJK-201717；7:ZJZSJK-201719；8:ZJZSJK-201722；9:
ZJZSJK-201724；10:ZJZSJK-201729.
Only there is salmonella species-specific amplification condition, it was demonstrated that this is salmonella, is not occurred other non-specific
Property amplified band.
2.2.3 five weight PCR detect the specificity of other bacteriums
To vibrio parahemolyticus, staphylococcus aureus, Listeria monocytogenes, Vibrio vulnificus, Shigella flexneri, Escherichia coli,
Vibrio mimicus, sonnei shigella dysenteriae, staphylococcus epidermis, enterococcus, citrobacter freundii are 12 kinds of Klebsiella Pneumoniae etc. thin
The amplification of bacterium genomic DNA is feminine gender.See Fig. 3, wherein M:DL1000；1: five heavy PCR specific amplification band
(221bp,342bp,532bp,656bp,856bp)；2: vibrio parahemolyticus；3: staphylococcus aureus；4: single to increase Liszt
Bacterium；5: Vibrio vulnificus；6: Shigella flexneri；7: Escherichia coli；8: vibrio mimicus；9: sonnei shigella dysenteriae；10: epidermis grape ball
Bacterium；11: enterococcus；12: citrobacter freundii；13: Klebsiella Pneumoniae.
From the above verification result it is known that the five weight PCR that the present invention establishes, which detect 4 kinds, does not have to serotype salmonella tool
There is fine specificity.The present invention can not only detect whether as salmonella, and can be just quasi- simultaneously by PCR detection merely
Really identify whether be salmonella typhimurium, Bacterium enteritidis, salmonella typhi, Salmonella choleraesuls, without borrowing
Help other ancillary technique means.
Raw materials used in the present invention, equipment is unless otherwise noted the common raw material, equipment of this field；In the present invention
Method therefor is unless otherwise noted the conventional method of this field.
The above is only presently preferred embodiments of the present invention, is not intended to limit the invention in any way, it is all according to the present invention
Technical spirit any simple modification, change and equivalent transformation to the above embodiments, still fall within the technology of the present invention side
The protection scope of case.
Sequence table
<110>complex art service centre, Zhoushan Entry-Exit Inspection and Quarantine Bureau
<120>a kind of five heavy PCR primers, kit and its application for detecting four kinds of Salmonella serogroups
<130> 2018
<160> 10
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 1
ggacttgccg actggttaat 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 2
aatactgcgc tgccagatag 20
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 3
tggttggttc gtcactgatt 20
<210> 4
<211> 23
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 4
ctacgttcgt tcttctggta ctt 23
<210> 5
<211> 23
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 5
cacctccact actacaagga aac 23
<210> 6
<211> 20
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 6
gggtggcaag ggaatgaata 20
<210> 7
<211> 22
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 7
gttcattggc aaggtcgtta tt 22
<210> 8
<211> 22
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 8
gggctttcta tcgagggatt ag 22
<210> 9
<211> 21
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 9
cctgacaacg aagggtgatt t 21
<210> 10
<211> 20
<212> DNA
<213>artificial sequence (Artificial sequence)
<400> 10
agtcgctctt tcccgtatct 20
Claims (4)
1. a kind of five heavy PCR primers for detecting four kinds of Salmonella serogroups, including five pairs of specific primers, it is characterised in that:
Five pairs of specific primers are respectively as follows:
Salmonella-F, sequence information is as shown in SEQ ID No.1；
Salmonella-R, sequence information is as shown in SEQ ID No.2；
Bacterium enteritidis-F, sequence information is as shown in SEQ ID No.3；
Bacterium enteritidis-R, sequence information is as shown in SEQ ID No.4；
Salmonella typhimurium-F, sequence information is as shown in SEQ ID No.5；
Salmonella typhimurium-R, sequence information is as shown in SEQ ID No.6；
Salmonella typhi-F, sequence information is as shown in SEQ ID No.7；
Salmonella typhi-R, sequence information is as shown in SEQ ID No.8；
Salmonella choleraesuls-F, sequence information is as shown in SEQ ID No.9；
Salmonella choleraesuls-R, sequence information is as shown in SEQ ID No.10.
2. a kind of kit using five weights PCR primer as described in claim 1, which is characterized in that including five heavy PCR reactants
System, the five weights PCR reaction system are contained in terms of 50 μ L of total volume:
2 × Multiplex PCR Mix1,25 μ L,
0.25 μ L of Multiplex PCR Mix2,
0.4 μM of salmonella-F,
0.4 μM of salmonella-R,
0.4 μM of Bacterium enteritidis-F,
0.4 μM of Bacterium enteritidis-R,
0.4 μM of salmonella typhimurium-F,
0.4 μM of salmonella typhimurium-R,
0.4 μM of salmonella typhi-F,
0.4 μM of salmonella typhi-R,
0.4 μM of Salmonella choleraesuls-F,
0.4 μM of Salmonella choleraesuls-R,
4 μ L of template,
Moisturizing is to 50 μ L.
3. a kind of application detected in four kinds of Salmonella serogroups using kit as claimed in claim 2 in PCR.
4. application as claimed in claim 3, which is characterized in that PCR amplification condition are as follows: 94 DEG C of 1min；94℃30sec；57℃
30sec；72℃ 45sec；35 circulations, 72 DEG C of 10min；4℃；1 circulation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811462307.8A CN109468394A (en) | 2018-11-30 | 2018-11-30 | A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811462307.8A CN109468394A (en) | 2018-11-30 | 2018-11-30 | A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109468394A true CN109468394A (en) | 2019-03-15 |
Family
ID=65674737
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811462307.8A Pending CN109468394A (en) | 2018-11-30 | 2018-11-30 | A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109468394A (en) |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008161170A (en) * | 2007-01-04 | 2008-07-17 | Daikin Ind Ltd | Oligonucleotide for highly-sensitive detection of bacterium of genus salmonella and detection method and detection kit each using the same |
| CN101565737A (en) * | 2008-04-21 | 2009-10-28 | 于维森 | Fast check reagent of Salmonella causing food poisoning |
| WO2013007996A1 (en) * | 2011-07-08 | 2013-01-17 | Health Protection Agency | Salmonella detection assay |
| CN103320507A (en) * | 2013-07-01 | 2013-09-25 | 黑龙江出入境检验检疫局检验检疫技术中心 | DPO primer sequences for salmonella detection by using DPO-PCR method, and detection kit thereof |
| CN104059977A (en) * | 2014-06-25 | 2014-09-24 | 上海交通大学 | Salmonella serotype identification method and kit thereof |
| CN104087654A (en) * | 2013-04-01 | 2014-10-08 | 中国农业大学 | Multiple PCR identification kit of salmonella and five serotypes of salmonella |
| CN105154530A (en) * | 2015-07-23 | 2015-12-16 | 舟山出入境检验检疫局综合技术服务中心 | Eightfold PCR amplification kit and amplification primer thereof for detecting pathogenic bacteria in food |
-
2018
- 2018-11-30 CN CN201811462307.8A patent/CN109468394A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008161170A (en) * | 2007-01-04 | 2008-07-17 | Daikin Ind Ltd | Oligonucleotide for highly-sensitive detection of bacterium of genus salmonella and detection method and detection kit each using the same |
| CN101565737A (en) * | 2008-04-21 | 2009-10-28 | 于维森 | Fast check reagent of Salmonella causing food poisoning |
| WO2013007996A1 (en) * | 2011-07-08 | 2013-01-17 | Health Protection Agency | Salmonella detection assay |
| CN104087654A (en) * | 2013-04-01 | 2014-10-08 | 中国农业大学 | Multiple PCR identification kit of salmonella and five serotypes of salmonella |
| CN103320507A (en) * | 2013-07-01 | 2013-09-25 | 黑龙江出入境检验检疫局检验检疫技术中心 | DPO primer sequences for salmonella detection by using DPO-PCR method, and detection kit thereof |
| CN104059977A (en) * | 2014-06-25 | 2014-09-24 | 上海交通大学 | Salmonella serotype identification method and kit thereof |
| CN105154530A (en) * | 2015-07-23 | 2015-12-16 | 舟山出入境检验检疫局综合技术服务中心 | Eightfold PCR amplification kit and amplification primer thereof for detecting pathogenic bacteria in food |
Non-Patent Citations (2)
| Title |
|---|
| SI HONG PARK等: "Identification of Salmonella enteric subspecies I, Salmonella enteric serovars Typhimurium, Enteritidis and Typhi using multiplex PCR", 《FEMS MICROBIOL LETT》 * |
| 郭华麟: "双启动寡核苷酸聚合酶链式反应法检测食品中的沙门氏菌", 《食品安全质量检测学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Oniciuc et al. | The present and future of whole genome sequencing (WGS) and whole metagenome sequencing (WMS) for surveillance of antimicrobial resistant microorganisms and antimicrobial resistance genes across the food chain | |
| Sung et al. | Comparison of gastric microbiota between gastric juice and mucosa by next generation sequencing method | |
| Wei et al. | Simultaneous detection of Escherichia coli O157: H7, Staphylococcus aureus and Salmonella by multiplex PCR in milk | |
| Laing et al. | Pan-genome analyses of the species Salmonella enterica, and identification of genomic markers predictive for species, subspecies, and serovar | |
| Van Immerseel et al. | Salmonella Gallinarum field isolates from laying hens are related to the vaccine strain SG9R | |
| Roussel et al. | Comparison of conventional plating, PMA-qPCR, and flow cytometry for the determination of viable enterotoxigenic Escherichia coli along a gastrointestinal in vitro model | |
| CN105648055B (en) | A kind of fowl source salmonella multiple PCR detection kit and its nondiagnostic detection method | |
| CN105936935B (en) | PCR detection kit for rapidly identifying specific serotype salmonella | |
| Liang et al. | Comparison of the filtration culture and multiple real-time PCR examination for Campylobacter spp. from stool specimens in diarrheal patients | |
| Lee et al. | Antibiotic susceptibility, genetic diversity, and the presence of toxin producing genes in Campylobacter isolates from poultry | |
| CN107365869B (en) | Method and primer for detecting Klebsiella pneumoniae by loop-mediated isothermal amplification technology | |
| Silva et al. | Genomic and phenotypic characterization of Campylobacter fetus subsp. venerealis strains | |
| CN115786543A (en) | Multiplex PCR detection kit for identifying and distinguishing salmonella pullorum and salmonella gallinarum | |
| Badr et al. | Phenotypic and genotypic screening of colistin resistance associated with emerging pathogenic Escherichia coli isolated from poultry | |
| Sánchez-Alonzo et al. | Nutrient deficiency promotes the entry of Helicobacter pylori cells into candida yeast cells | |
| Akter et al. | Prevalence of Shigella boydii in Bangladesh: isolation and characterization of a rare phage MK-13 that can robustly identify shigellosis caused by Shigella boydii type 1 | |
| Zhang et al. | Simple and rapid detection of Salmonella by direct PCR amplification of gene fimW | |
| Rani et al. | Point-of-care lateral flow detection of viable Escherichia coli O157: H7 using an improved propidium monoazide-recombinase polymerase amplification method | |
| CN106244690B (en) | A kind of multiple PCR detection kit of Rapid identification Salmonella enteritidis, white diarrhea/Salmonella gallinarum and Salmonella dublin | |
| CN104278102B (en) | The Quadruple-PCR detection method of a kind of salmonella | |
| CN109468394A (en) | A kind of five heavy PCR primers, kit and its application detecting four kinds of Salmonella serogroups | |
| CN104531860B (en) | A kind of molecular detecting method of Shigella and its application | |
| Odyniec et al. | Assessment of the Role of Free-Living and Farmed Fallow Deer (Dama dama) as A Potential Source of Human Infection with Multiple-Drug-Resistant Strains of Yersinia enterocolitica and Yersinia pseudotuberculosis | |
| AU2019100071A4 (en) | Pcr detection kit for rapidly identifying salmonella of specific serotypes | |
| Kheiri et al. | Multiplex PCR for detection of water-borne bacteria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190315 |
|
| RJ01 | Rejection of invention patent application after publication |