CN110257540A - Detect PCR primer, primer sets, probe and its kit and detection method of Proteus and proteus mirabilis - Google Patents
Detect PCR primer, primer sets, probe and its kit and detection method of Proteus and proteus mirabilis Download PDFInfo
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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Abstract
The present invention is suitable for Bacteria Detection technical field, provide a kind of PCR primer, primer sets, probe and its kit and detection method for detecting Proteus and proteus mirabilis, disclose the nucleotide sequence of 4 PCR primers of detection Proteus and proteus mirabilis, totally 2 pairs, kit comprising the primer sets, the characteristic with high specificity, simultaneously, high sensitivity, detection lower bound are 102CFU/mL, testing result are accurate and reliable；Without carrying out electrophoresis detection to target gene amplified production, it is easy to operate, detection cycle is short, the workload of operator can be reduced, obtains accurate detection information in time, enormously simplifies the process of proteus detection, shorten detection cycle, it can be used as sensitive primary dcreening operation means, improve the positive rate of proteus culture detection, proteus Hygienic Index in food can be formulated from now on for China, strong technical support is provided.
Description
Technical field
The invention belongs to Bacteria Detection technical field more particularly to a kind of detection Proteus and proteus mirabilises
PCR primer, primer sets, probe and its kit and detection method.
Background technique
Proteus is no pod membrane, without gemma, amphitrichous and fimbriate gram-Negative bacillus, is distributed widely in certainly
It is that the essential condition for causing humans and animals to infect is caused a disease so in the enteron aisle of boundary, animal wastes, clinical samples and humans and animals
Bacterium.In recent years, Proteus Infection fashion trend constantly expands.Caused in food according to related Proteus reported in the literature
The statistical result of poison shows that Proteus food poisoning in China's is in rise year by year trend.In addition to causing food poisoning, deformed rod
Pseudomonas can also cause multi-infection, as respiratory tract infection, urethral infection, wound and infection of burn, pneumonia, enteritis, it is chronic in
Otitis, meningitis, peritonitis, septicemia, rheumathritis, pulmonary tuberculosis etc..Wherein Proteus be urethral infection most
One of the main pathogenic fungi.Therefore, enough attention should be caused to the quick diagnosis of Proteus and prevention and control.
Currently, the quantitative detection method of proteus relies primarily on traditional biochemical identification method and regular-PCR method both at home and abroad,
Conventional method will by Zengjing Granule, isolate and purify, biochemical reaction, the processes such as serological test, there are detection process operation is numerous
It is trivial, it takes a long time, spends high, the requirement height to culture medium, there are larger for the sensitivity detected between the different laboratory of condition
Otherness, be not easy evaluation and quality control the disadvantages of.Although the detection of regular-PCR technology is accurate, quick and cost is relatively low, mesh
Gene magnification after need electrophoresis, the health of time-consuming and easy damage testing staff.
It can be seen that the quantitative detection method of the proteus of the prior art there are it is cumbersome, take a long time, detection spirit
Sensitivity is low and security hidden trouble.
Summary of the invention
The PCR primer for being designed to provide detection Proteus and proteus mirabilis of the embodiment of the present invention, it is intended to
Solve the quantitative detection method of proteus of the prior art there are it is cumbersome, take a long time, detection sensitivity is low and peace
Full potential problem.
The embodiments of the present invention are implemented as follows, detects the PCR primer of Proteus and proteus mirabilis, primer sequence
It is classified as SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4；Or the nucleosides for the primer sequence
Sour complementary series；
The nucleotide sequence of the primer is as follows:
SEQ ID NO:1 AAGTATACGAAGCGTCCCTACG,
SEQ ID NO:2 CGTCTGCCACGGCTTAAAC,
SEQ ID NO:3 CGGGTACCCTGTGGTAATG,
SEQ ID NO:4 CTGTTCGCGGATTATTTGTT。
The another object of the embodiment of the present invention is to detect the PCR primer group of Proteus and proteus mirabilis, draw
Object group is primer sets 1 or primer sets 2；
Wherein, primer sets 1 are made of primer sequence SEQ ID NO:1 and SEQ ID NO:2；Or by the primer sequence
The nucleotide complementary series of column forms；
Primer sets 2 are made of primer sequence SEQ ID NO:3 and SEQ ID NO:4；Or by the primer sequence
Nucleotide complementary series composition.
The another object of the embodiment of the present invention is the PCR detection probe of detection Proteus and proteus mirabilis,
Probe sequence is as follows:
SEQ ID NO:5 ACAGTTAAAGTTCAGCAGGCGGTGGTAT,
SEQ ID NO:6 TACCCCACAGTCCCGGTATTTGGAA；
Or the nucleotide complementary series for the probe sequence.
The another object of the embodiment of the present invention is to detect the PCR detection reagent of Proteus and proteus mirabilis
Box contains above-mentioned PCR primer group.
The another object of the embodiment of the present invention is the PCR detection method of detection Proteus and proteus mirabilis,
Include:
Obtain the template DNA of sample to be tested；
QPCR SuperMix, primer sets, probe, template DNA and H2O is anti-according to scheduled preparation method preparation PCR
Answer system；
After the PCR reaction system mixing centrifugation, PCR reaction is carried out under preset amplification program, and fixed based on fluorescence
The measurement that PCR instrument carries out fluorescence intensity is measured, to detect to Proteus and proteus mirabilis.
For the embodiment of the present invention using Proteus and proteus mirabilis as aimed strain, deformation can be detected simultaneously by establishing
The PCR primer of Bacillus and proteus mirabilis, primer sets, probe and its kit and detection method, with high specificity
Characteristic, meanwhile, high sensitivity, detection lower bound is 102CFU/mL, testing result are accurate and reliable；It is produced without being expanded to target gene
Object carries out electrophoresis detection, and easy to operate, detection cycle is short, it is possible to reduce the workload of operator obtains accurate in time
Detection information enormously simplifies the process of proteus detection, shortens detection cycle, can be used as sensitive primary dcreening operation means, improves
The positive rate of proteus culture detection, proteus Hygienic Index in food can be formulated from now on for China and provides strong skill
Art support.
Detailed description of the invention
Fig. 1 is the specific test figure of Proteus provided in an embodiment of the present invention and proteus mirabilis；
Fig. 2 is the standard curve of fluorescent PCR amplification Proteus provided in an embodiment of the present invention；
Fig. 3 is the standard curve of fluorescent PCR amplification proteus mirabilis provided in an embodiment of the present invention.
Specific embodiment
In order to more fairly set out technology contents of the invention, it is described in detail in conjunction with specific embodiments herein, it is clear that
Cited embodiment is the preferred embodiment of the technical program, and those skilled in the art can be according to disclosed skill
The other technologies scheme that art content is apparent from still falls within protection scope of the present invention.
Embodiment 1 detects the PCR primer group of Proteus and proteus mirabilis
The embodiment of the present invention is by Proteus (Proteus), proteus mirabilis (Proteus mirabilis) and 10
The non-targeted bacterium of kind (Escherichia coli, single increasing listeria spp, slope bacillus, comma bacillus, Vibrio vulnificus, Vibrio flurialis, parahemolyticas arc
Bacterium, staphylococcus aureus, campylobacter jejuni, salmonella) genomic dna sequence be compared, finally determine deformed rod
The specific gene of Pseudomonas is atpD gene, and the specific gene of proteus mirabilis is ureC gene.
Further, in the target area of Proteus and proteus mirabilis, pass through design of primers principle and a variety of meters
Calculation machine program, design obtain the PCR primer group of Proteus and proteus mirabilis, and are filtered out both by many experiments
It is guarded in certifiable kind, and can guarantee the special primer sequence of inter-species, including 2 groups, nucleotide sequence distinguishes following institute
Show:
Primer sets 1:
SEQ ID NO:1 AAGTATACGAAGCGTCCCTACG,
SEQ ID NO:2 CGTCTGCCACGGCTTAAAC,
Or the nucleotide complementary series for the primer sequence；
Primer sets 2:
SEQ ID NO:3 CGGGTACCCTGTGGTAATG,
SEQ ID NO:4 CTGTTCGCGGATTATTTGTT,
Or the nucleotide complementary series for the primer sequence.
Embodiment 2 detects the PCR detection kit of Proteus and proteus mirabilis
The embodiment of the invention provides it is a kind of detection Proteus and proteus mirabilis PCR detection kit,
Including following component:
1) containing the primer sets in embodiment 1；
2) detection probe: probe sequence is as follows:
SEQ ID NO:5 ACAGTTAAAGTTCAGCAGGCGGTGGTAT,
SEQ ID NO:6 TACCCCACAGTCCCGGTATTTGGAA；
Or the nucleotide complementary series for the probe sequence.
The fluorophor of the end of probe sequence 5 ' label is one of FAM, VIC；The end label of probe sequence 3 ' is quenched
The group that goes out is BHQ1.
3) PCR reaction system:
Primer and probe is dissolved into 100uM, is diluted to 10uM.Respectively into 20uL system be added primer 0.6uL,
0.3uL, probe 0.3uL, 0.15uL, template DNA (10-100ng) 1uL, 3uL.QPCR SuperMix 10.0uL is constant.Through excellent
After change, determine that PCR reaction system (20 μ L) is as follows:
QPCR SuperMix 10.0uL, primer (10uM) each 0.3uL, probe (10uM) each 0.15uL, template DNA (10-
100ng) 3uL, H2O 5.5uL；
Wherein, qPCR SuperMix is purchase gained comprising Taq enzyme, PCR buffer, triphosphoric acid deoxyribose core
Glycosides, MgCl2。
Embodiment 3 detects the PCR detection method of Proteus and proteus mirabilis
The embodiment of the invention provides a kind of detection Proteus and the PCR detection methods of proteus mirabilis, utilize
The PCR detection kit that embodiment 2 is established, detects sample to be tested, steps are as follows:
Obtain the template DNA of sample to be tested:
1, the cell of 1.0~5.0E+09 is collected with 1.5ml centrifuge tube, 12000rmp is centrifuged 2 minutes, abandons supernatant (cell
Culture solution)；Be added Buffer GL of 180 μ L, the Proteinase K (Proteinase K) of 20 μ L and 10 μ L RNaseA (10mg,
ML) (ribonuclease A), sufficiently inhale play mixing, in 56 DEG C water-bath warm bath 10 minutes；The Buffer GB and 200 μ of 200 μ L is added
100% ethyl alcohol of L is sufficiently inhaled and plays mixing；
2. centrifugal column is placed on collecting pipe, solution is moved in centrifugal column, and 12000rpm is centrifuged 2 minutes, abandons filtrate.
3. being added the BufferWA of 500 μ L into centrifugal column, 12000rpm is centrifuged 1 minute, abandons filtrate.
4. being added the BufferWB of 700 μ L into centrifugal column, 12000rpm is centrifuged 1 minute, abandons filtrate.
5. repeating previous step.
6. centrifugal column is placed on collecting pipe, 12000rpm is centrifuged 2 minutes.
7. centrifugal column is placed on the centrifuge tube of new 1.5mL, it is added 50~200 μ L's in the centre of centrifugal column film
Aqua sterilisa or elution buffer are stored at room temperature 5 minutes.
8. 12000rpm is centrifuged 2 minutes and elutes template DNA.
The preparation of PCR reaction system:
PCR reaction system is prepared in sterilizing PCR pipe according to following system, wherein 20 μ LPCR reaction system configuration proportions
It is as follows:
QPCR SuperMix 10.0uL, primer (10uM) each 0.3uL, probe (10uM) each 0.15uL, template DNA (10-
100ng) 3uL, H2O 5.5uL。
PCR amplification program:
It is placed in PCR thermal cycler after PCR pipe is mixed centrifugation, following amplification program is set and starts PCR reaction,
Amplification program are as follows: the first step, 94 DEG C of thermal startings and 3~5min of initial denaturation；Second step, 94 DEG C of 15~30s of denaturation；60 DEG C of third step
45~60s of annealing and extension；Repeat second step, third step totally 40~45 circulations.
After the PCR reaction system mixing centrifugation, PCR reaction is carried out under preset amplification program, and fixed based on fluorescence
The measurement that PCR instrument carries out fluorescence intensity is measured, to detect to Proteus and proteus mirabilis.
4 specific test of embodiment
By proteus vulgaris, proteus mirabilis and 10 kinds of non-targeted bacterium (Escherichia coli, single increasing listeria spp, slope
Bacillus, comma bacillus, Vibrio vulnificus, Vibrio flurialis, vibrio parahemolyticus, staphylococcus aureus, campylobacter jejuni, salmonella
DNA is extracted after culture, carries out specific test with the determining amplification system of optimization and amplification program, as the result is shown: common variation
Bacillus and proteus mirabilis are in typical S type amplification curve, other 10 kinds of non-targeted bacterium no cross reactions (as shown in Figure 1).
Judgment basis: Ct<35.0 are judged to the positive, and Ct>35.0 are judged to feminine gender.
Therefore, according to specific test result it is found that the primer and probe that the present invention designs can expand Proteus
And proteus mirabilis, 10 kinds of non-targeted bacterium (Escherichia coli, it is single increase listeria spp, slope bacillus, comma bacillus, Vibrio vulnificus,
Vibrio flurialis, vibrio parahemolyticus, staphylococcus aureus, campylobacter jejuni, salmonella) it does not expand, show that the present invention is set
The primed probe of meter has good specificity.
The detection method for being separately cultured the proteus positive and negative sample verifying embodiment 3 of embodiment 5
The embodiment of the present invention is separately cultured the proteus positive and 20 parts of diarrhea patient excrement using 55 parts of diarrhea patient excrement
Just it is separately cultured the accuracy of the PCR detection method of proteus feminine gender verifying detection Proteus, wherein 75 parts of excrement marks
Originally both from the adult diarrhea patient of the food origin disease monitoring project randomly selected.
Stool sample extracts the step of nucleic acid:
1, excrement 180-220mg is weighed in 2mL centrifuge tube.
2,1mL InhibitEX Buffer is added, whirlpool shakes 1min to mixing well.
3,70 DEG C of incubation 5min.14000rpm is centrifuged 1min.
4,15uL Proteinase K is drawn in new 1.5mLEp pipe.
5, the supernatant 200uL drawn in 3 steps is added in the Ep pipe containing Proteinase K.
6, plus 200uL Buffer AL, vortex mix 15s.70 DEG C of incubation 10min.
7, add 200uL dehydrated alcohol, mix.
8, the liquid in 7 steps is carefully drawn in QIAapm centrifugal column, and 14000rpm is centrifuged 1min.Collecting pipe is abandoned, is replaced
The collecting pipe renewed.
9, plus 500uL Buffer AW1,14000rpm are centrifuged 1min.Collecting pipe is abandoned, new collecting pipe is replaced.
10, plus 500uL Buffer AW2,14000rpm are centrifuged 3min.Collecting pipe is abandoned, new collecting pipe is replaced.
11,14000rpm is centrifuged 3min.
12, QIAapm centrifugal column is transferred in new 1.5mLEp pipe, adds 200uL Buffer ATE, is placed at room temperature for
1min。
13,14000rpm is centrifuged 1min.
Using the DNA extracted in stool sample as template, fluorescence is carried out according to PCR reaction system above-mentioned and amplification program
Record is as a result, the Ct value for being separately cultured positive sample and ' negative ' specimens tested through fluorescence quantitative PCR instrument after PCR amplification
As shown in table 1-2.The results show that amplification is consistent with result is separately cultured.55 parts of proteus are separately cultured positive sample
In, 44 parts of sample multiple fluorescence PCR amplifications are positive, and coincidence rate reaches 80%.This reason may is that bacterium solution content in excrement
It is lower, although being separately cultured out proteus, cause fluorescent PCR that can not detect in the loss of DNA extraction process amplifying nucleic acid.20 parts
Proteus is separately cultured in negative sample, and 2 parts of sample multiple fluorescence PCR amplifications are positive.This two parts of samples expand through fluorescent PCR
It is sequenced after increasing, it was demonstrated that be proteus.The reason of isolated culture and multiple fluorescence PCR method have differences may are as follows: 1, multiple
The high sensitivity of fluorescent PCR method is in the method for conventional biochemical culture；2, there are the remaining nucleic acid of dead bacterium in sample.
Table 1 is separately cultured positive sample summary sheet
Table 2 is separately cultured ' negative ' specimens summary sheet
| Sample number | Proteus (FAM) | Proteus mirabilis (VIC) |
| Syq170 | —— | —— |
| Syq171 | —— | —— |
| Syq172 | —— | —— |
| Syq173 | —— | —— |
| Syq174 | 26.608 | 26.677 |
| Syq175 | —— | —— |
| Syq176 | —— | —— |
| Syq177 | 30.435 | 29.487 |
| Syq178 | —— | —— |
| Syq179 | —— | —— |
| Syq180 | —— | —— |
| Syq181 | —— | —— |
| Syq182 | —— | —— |
| Syq183 | —— | —— |
| Syq184 | —— | —— |
| Syq185 | —— | —— |
| Syq186 | —— | —— |
| Syq187 | —— | —— |
| Syq188 | —— | —— |
| Syq189 | —— | —— |
6 sensitivity test of embodiment
(1) with colony counting method to the concentration quantitative of proteus mirabilis bacteria suspension
A takes out proteus mirabilis from -80 DEG C of refrigerators, and after thawing, one ring of picking is coated on LB plate, 37 DEG C of trainings
It supports for 24 hours.
B picks a small amount of lawn in 10mL sterile saline with cotton swab, mixes.
C draws 100uL bacteria suspension and detects OD value, until OD600 value reaches 1.0.
When d has been generally acknowledged that OD600 value is 1.0, the content of bacterium is about 108CFU/mL.The 1OD bacteria suspension that c is measured is step by step
It dilutes (100uL bacterium solution+900uL physiological saline).
E takes 10 respectively4、103、102、101Bacterium solution 100uL be coated on LB plate, 37 DEG C culture for 24 hours, count.
(2) fluorescent PCR amplification Proteus and proteus mirabilis standard curve (as Figure 2-3)
A will dilute 108-101Bacterium solution be centrifuged 5min in 95 DEG C of incubation 10min, 12000rpm, Aspirate supernatant in
In new centrifuge tube.
B does two repetitions with a system 20uL, each concentration, adds respectively in centrifuge tube according to the amplification system optimized
Enter required premixed liquid qPCR SuperMix, high pressure sterilization water, primer, probe, mixing are sub-packed in microcentrifugal tube.
(3) it is centrifuged after the template being added in (2) a, amplification program (the 94 DEG C of 5min of initial denaturation optimized according to embodiment 3；
94 DEG C of 15s are denaturalized, anneal 60 DEG C of 1min, 40 circulations) carry out PCR amplification.
The detection lower bound of tests determined Proteus and proteus mirabilis is 102CFU/mL shows the present invention
The primer and probe high sensitivity of design.
To sum up, using Proteus and proteus mirabilis as aimed strain, establish to examine the embodiment of the present invention simultaneously
Survey PCR primer, primer sets, probe and its kit and detection method of Proteus and proteus mirabilis, on the one hand, this
Two pairs of primer and probe specific detection Proteus and proteus mirabilis are used in invention, and to other, non-targeted bacterium is (greatly in 10
Enterobacteria, single increasing listeria spp, slope bacillus, comma bacillus, Vibrio vulnificus, Vibrio flurialis, vibrio parahemolyticus, golden yellow grape
Coccus, campylobacter jejuni, salmonella) without amplification, the characteristic with high specificity, meanwhile, high sensitivity, detecting lower bound is
102CFU/mL, testing result are accurate and reliable；On the other hand, the present invention is not necessarily to carry out electrophoresis inspection to target gene amplified production
It surveys, easy to operate, detection cycle is short, it is possible to reduce the workload of operator obtains accurate detection information in time, significantly simple
Change the process of proteus detection, shortened detection cycle, can be used as sensitive primary dcreening operation means, improves proteus culture detection
Positive rate, proteus Hygienic Index in food can be formulated from now on for China, strong technical support is provided.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
Limitations on the scope of the patent of the present invention therefore cannot be interpreted as.It should be pointed out that for those of ordinary skill in the art
For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to guarantor of the invention
Protect range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Made any modifications, equivalent replacements, and improvements etc., should all be included in the protection scope of the present invention within mind and principle.
Sequence table
<110>Inst of Infection Disease Prevention and Control, Chinese Diseases Prevention an
<120>PCR primer, primer sets, probe and its kit and the detection side of Proteus and proteus mirabilis are detected
Method
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<170> SIPOSequenceListing 1.0
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aagtatacga agcgtcccta cg 22
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cgtctgccac ggcttaaac 19
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cgggtaccct gtggtaatg 19
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Claims (10)
1. detecting the PCR primer of Proteus and proteus mirabilis, which is characterized in that primer sequence be SEQ ID NO:1,
SEQ ID NO:2,SEQ ID NO:3,SEQ ID NO:4；Or the nucleotide complementary series for the primer sequence；
The nucleotide sequence of the primer is as follows:
2. detecting the PCR primer group of Proteus and proteus mirabilis, which is characterized in that primer sets are primer sets 1 or draw
Object group 2；
Wherein, primer sets 1 are made of primer sequence SEQ ID NO:1 and SEQ ID NO:2；Or by the primer sequence
Nucleotide complementary series composition；
Primer sets 2 are made of primer sequence SEQ ID NO:3 and SEQ ID NO:4；Or the nucleosides by the primer sequence
Sour complementary series composition.
3. detecting the PCR detection probe of Proteus and proteus mirabilis, which is characterized in that probe sequence is as follows:
SEQ ID NO:5 ACAGTTAAAGTTCAGCAGGCGGTGGTAT,
SEQ ID NO:6 TACCCCACAGTCCCGGTATTTGGAA；
Or the nucleotide complementary series for the probe sequence.
4. the PCR detection probe of detection Proteus according to claim 3 and proteus mirabilis, feature exists
In the fluorophor of the end of probe sequence 5 ' label is one of FAM, VIC；The quenching group of the end of probe sequence 3 ' label
For BHQ1.
5. the PCR detection kit of a kind of detection Proteus and proteus mirabilis, which is characterized in that contain claim
PCR primer group described in 2.
6. the PCR detection kit of a kind of detection Proteus according to claim 5 and proteus mirabilis, special
Sign is that the PCR detection kit also contains detection probe as claimed in claim 3.
7. the PCR detection kit of a kind of detection Proteus according to claim 5 and proteus mirabilis, special
Sign is that the PCR detection kit also contains: qPCR SuperMix, primer sets, probe, template DNA, H2O。
8. the PCR detection kit of a kind of detection Proteus according to claim 7 and proteus mirabilis, special
Sign is that the qPCR SuperMix includes Taq enzyme, PCR buffer, triphosphoric acid dezyribonucleoside, MgCl2。
9. the PCR detection method of a kind of detection Proteus and proteus mirabilis characterized by comprising
Obtain the template DNA of sample to be tested；
By qPCR SuperMix, primer sets, probe, template DNA and H2O prepares PCR reactant according to scheduled preparation method
System；
After the PCR reaction system mixing centrifugation, PCR reaction is carried out under preset amplification program, and be based on quantitative fluorescent PCR
Instrument carries out the measurement of fluorescence intensity, to detect to Proteus and proteus mirabilis.
10. the PCR detection method of a kind of detection Proteus according to claim 9 and proteus mirabilis, special
Sign is, the preset amplification program are as follows: 94 DEG C of 3~5min of initial denaturation；Be denaturalized 94 DEG C of 15~30s, annealing 60 DEG C 45~
60s, 40~45 circulations.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110551835A (en) * | 2019-10-16 | 2019-12-10 | 四川农业大学 | specific primer and method for detecting bovine proteus mirabilis |
| CN110951895A (en) * | 2019-12-24 | 2020-04-03 | 重庆市畜牧科学院 | System and method for detecting and distinguishing proteus mirabilis, proteus vulgaris and proteus pani |
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| CN110551835A (en) * | 2019-10-16 | 2019-12-10 | 四川农业大学 | specific primer and method for detecting bovine proteus mirabilis |
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| CN113512601A (en) * | 2021-06-30 | 2021-10-19 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Molecular target for screening proteus and quantitative detection method |
| CN113512601B (en) * | 2021-06-30 | 2023-10-20 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Molecular targets for screening for Proteus and quantitative detection methods |
| CN114214440A (en) * | 2021-12-14 | 2022-03-22 | 广州双螺旋基因技术有限公司 | LAMP detection kit for detecting proteus mirabilis |
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