CN110643723A - Primer group for detecting ureaplasma parvum - Google Patents
Primer group for detecting ureaplasma parvum Download PDFInfo
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- CN110643723A CN110643723A CN201910915087.8A CN201910915087A CN110643723A CN 110643723 A CN110643723 A CN 110643723A CN 201910915087 A CN201910915087 A CN 201910915087A CN 110643723 A CN110643723 A CN 110643723A
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- ureaplasma
- primer
- primer group
- detecting
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- 241000935255 Ureaplasma parvum Species 0.000 title claims abstract description 18
- 240000008791 Antiaris toxicaria Species 0.000 claims abstract description 8
- 108010046334 Urease Proteins 0.000 claims abstract description 8
- 241000202898 Ureaplasma Species 0.000 abstract description 13
- 238000001514 detection method Methods 0.000 abstract description 11
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 244000052769 pathogen Species 0.000 abstract description 2
- 230000001717 pathogenic effect Effects 0.000 abstract description 2
- 241000202921 Ureaplasma urealyticum Species 0.000 description 12
- 239000000523 sample Substances 0.000 description 8
- 238000012408 PCR amplification Methods 0.000 description 4
- 241000204031 Mycoplasma Species 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 230000037029 cross reaction Effects 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 206010046367 Ureaplasma infections Diseases 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241001430197 Mollicutes Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Images
Classifications
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- 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
Abstract
The invention relates to the technical field of pathogen detection, and discloses a primer group for detecting ureaplasma parvum, which comprises a primer group of ureaplasma parvum urease genes, wherein the name of a forward primer is UpS, and the name of a reverse primer is UPAS. The primer group can be specially used for detecting the small ureaplasma, can accurately detect the small ureaplasma, has low requirement on detection equipment, low cost, easy realization and high detection sensitivity and accuracy, and does not generate cross reaction.
Description
Technical Field
The invention relates to the technical field of pathogen detection, in particular to a primer group for detecting ureaplasma parvum.
Background
Ureaplasma urealyticum (Ureaplasma) is one of 13 mycoplasma isolated from human body and capable of self-reproduction, and belongs to the genus Ureaplasma of the family mycoplasma of the order mycoplasma of the class mollicutes of the phylum firmicutes. Ureaplasma is intermediate between viruses and bacteria, is the smallest prokaryotic cell type microorganism that can reproduce on artificial media, has no cell wall, can self-replicate, is in the shape of a sphere and has high polymorphism. The Ureaplasma urealyticum is divided into 2 biological groups and 14 serotypes according to the molecular biological characteristics, the two biological groups comprise Ureaplasma parvum (Up) and Ureaplasma urealyticum (Uu), and of the 14 serotypes, serotypes 1, 3, 6 and 14 with smaller genome belong to the Ureaplasma parvum and account for 90 to 92 percent of Ureaplasma infection; serotypes 2, 4, 5 and 8-13 with larger genomes belong to ureaplasma urealyticum and account for 8-10% of ureaplasma infection.
The distribution of the small ureaplasma and the ureaplasma urealyticum in people and the sensitivity to drugs are different, and the common ureaplasma serotyping method is more complicated in operation, has higher requirements on equipment, is easy to generate cross reaction, and is difficult to accurately judge the result.
Therefore, in order to overcome the disadvantages and shortcomings of the prior art, the present invention intends to provide a primer set for accurately detecting ureaplasma parvum, so as to avoid cross-reaction and improve the accuracy of the detection result.
Disclosure of Invention
Based on the problems, the invention provides the primer group for detecting the small ureaplasma, the primer group can accurately detect the small ureaplasma, and the primer group has the advantages of high sensitivity and accuracy, low requirement on equipment and strong practicability.
In order to solve the technical problems, the invention provides the following technical scheme:
the primer group for detecting the ureaplasma parvum comprises a primer group of ureaplasma parvum urease genes, wherein the primer group comprises a forward primer UpS and a reverse primer UpAS, and the sequences of UpS and UpAS are as follows:
compared with the prior art, the invention has the beneficial effects that: the primer group can be specially used for detecting the small ureaplasma, can accurately detect the small ureaplasma, has low requirement on detection equipment, low cost, easy realization and high detection sensitivity and accuracy, and does not generate cross reaction.
Drawings
FIG. 1 is a diagram showing the results of electrophoresis obtained by detecting a primer set for a urease gene of ureaplasma parvum in an example of the present invention;
FIG. 2 is a diagram showing the results of electrophoresis obtained by detecting the primer set of the urease gene of ureaplasma urealyticum in the example of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples and accompanying drawings, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not meant to limit the present invention.
Example (b):
the primer group for detecting the ureaplasma parvum comprises a primer group of ureaplasma parvum urease genes, wherein the primer group comprises a forward primer UpS and a reverse primer UpAS, and the sequences of UpS and UpAS are as follows:
in this embodiment, a set of primer sets for detecting ureaplasma urealyticum is further selected, the primer sets include a forward primer UuS and a reverse primer UuAS, the primer sets used for detection are primers of urease genes of ureaplasma urealyticum, and the sequences of the UuS and the UuAS are as follows:
in this embodiment, a clinical sample containing both ureaplasma parvum and ureaplasma urealyticum is selected, the sample is equally divided into two groups, i.e., a group a and a group B, and then the group a sample is detected by using the above-mentioned ureaplasma parvum primer group, and the group B sample is detected by using the ureaplasma urealyticum primer group, the detection method includes the following steps:
s1: separately extracting sample DNA of group A and group B
Extracting sample DNA of group A and group B using a TIANAmp Genomic DNA Kit, wherein the Kit may be any one of those for extracting nucleic acid;
s2: PCR amplification of sample DNA
The PCR amplification system is shown below:
the specific primer-as is UuAS or UpAS, and the specific primer-s is UuS or UpS, in this example, two sets of the PCR reaction systems are respectively configured to be used for detecting ureaplasma micans and ureaplasma urealyticum in the sample, and the reaction procedure of the PCR amplification is as follows: pre-denaturation at 94 ℃ for 3 min; denaturation at 94 ℃ for 15s, annealing at 60 ℃ for 30s, and extension at 72 ℃ for 30s for 35 cycles; finally, extension is carried out for 7min at 72 ℃;
s3: detection of amplified bands by agarose gel electrophoresis
After the PCR amplification in step S2 was completed, a sample solution was prepared in the following proportions:
after the preparation of the above-mentioned sample solution was completed, 2% agarose gel electrophoresis was performed, and the position and brightness of the objective band were observed, the voltage of the agarose gel electrophoresis was 100V, and the electrophoresis time was 18 min.
The results of the two groups of primer groups after being respectively detected are shown in the attached figures 1 and 2, the attached figure 1 is a result graph obtained by using the primer group of the ureaplasma micrum urease gene, the attached figure 2 is a result graph obtained by using the primer group of the ureaplasma urealyticum, and the strips of the detection result in the attached figure 1 are clear, the boundary is clear, and the situations of dragging and mixing do not exist.
The above is an embodiment of the present invention. The embodiments and specific parameters in the embodiments are only for the purpose of clearly illustrating the verification process of the invention and are not intended to limit the scope of the invention, which is defined by the claims, and all equivalent structural changes made by using the contents of the specification and the drawings of the present invention should be covered by the scope of the present invention.
Sequence listing
<110> Liujun
<120> primer set for detecting ureaplasma parvum
<130> 20190910
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 22
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
caggatcatc aagtcaattt ag 22
<210> 2
<211> 23
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
gataaaaagg ggaacattat gtt 23
Claims (1)
1. The primer group for detecting the ureaplasma parvum is characterized by comprising a primer group of a ureaplasma parvum urease gene, wherein the primer group comprises a forward primer UpS and a reverse primer UpAS, and the sequences of UpS and UpAS are as follows:
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910915087.8A CN110643723A (en) | 2019-09-26 | 2019-09-26 | Primer group for detecting ureaplasma parvum |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910915087.8A CN110643723A (en) | 2019-09-26 | 2019-09-26 | Primer group for detecting ureaplasma parvum |
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| Publication Number | Publication Date |
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| CN110643723A true CN110643723A (en) | 2020-01-03 |
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| CN201910915087.8A Pending CN110643723A (en) | 2019-09-26 | 2019-09-26 | Primer group for detecting ureaplasma parvum |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050244836A1 (en) * | 2004-04-28 | 2005-11-03 | Tat-Kin Tsang | Methods and compositions to detect bacteria using multiplex PCR |
| US20070037181A1 (en) * | 2005-02-17 | 2007-02-15 | Hovsep Melkonyan | Compositions and methods for detecting pathogen specific nucleic acids in urine |
| WO2014088088A1 (en) * | 2012-12-07 | 2014-06-12 | 三菱化学メディエンス株式会社 | Method for detecting and identifying bacterium belonging to genus mycoplasma or ureaplasma |
| CN103882133A (en) * | 2014-03-31 | 2014-06-25 | 深圳意达凯生物科技有限公司 | Primer pair for detecting ureaplasma urealyticum, kit and application thereof |
-
2019
- 2019-09-26 CN CN201910915087.8A patent/CN110643723A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050244836A1 (en) * | 2004-04-28 | 2005-11-03 | Tat-Kin Tsang | Methods and compositions to detect bacteria using multiplex PCR |
| US20070037181A1 (en) * | 2005-02-17 | 2007-02-15 | Hovsep Melkonyan | Compositions and methods for detecting pathogen specific nucleic acids in urine |
| WO2014088088A1 (en) * | 2012-12-07 | 2014-06-12 | 三菱化学メディエンス株式会社 | Method for detecting and identifying bacterium belonging to genus mycoplasma or ureaplasma |
| CN103882133A (en) * | 2014-03-31 | 2014-06-25 | 深圳意达凯生物科技有限公司 | Primer pair for detecting ureaplasma urealyticum, kit and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| FANRONG KONG 等: "Species Identification and Subtyping of Ureaplasma parvum and Ureaplasma urealyticum Using PCR-Based Assays", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
| MAŁGORZATA BIERNAT-SUDOLSKA等: "ssessment of various diagnostic methods of ureaplasma respiratory tract infections in newborns", 《DIAGNOSTIC METHODS OF UREAPLASMA INFECTIONS》 * |
| SHARLENE GOVENDER: "Epidemiology and Antibiotic Susceptibility Patterns of Mycoplasmasp. and Ureaplasma urealyticum", 《DISSERTATION PRESENTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY IN MEDICAL MICROBIOLOGY AT THE UNIVERSITY OF STELLENBOSCH》 * |
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