CN110734993A - nucleic acid detection test paper strip for distinguishing Brucella vaccine strain S19 from naturally infected strain - Google Patents
nucleic acid detection test paper strip for distinguishing Brucella vaccine strain S19 from naturally infected strain Download PDFInfo
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- 241000589562 Brucella Species 0.000 title claims abstract description 96
- 229960005486 vaccine Drugs 0.000 title claims abstract description 56
- 238000001514 detection method Methods 0.000 title claims abstract description 38
- 108020004707 nucleic acids Proteins 0.000 title claims description 21
- 102000039446 nucleic acids Human genes 0.000 title claims description 21
- 150000007523 nucleic acids Chemical class 0.000 title claims description 20
- 208000015181 infectious disease Diseases 0.000 claims abstract description 6
- 239000000523 sample Substances 0.000 claims description 27
- 206010006500 Brucellosis Diseases 0.000 abstract description 6
- 238000011330 nucleic acid test Methods 0.000 abstract description 2
- 244000052769 pathogen Species 0.000 abstract description 2
- 230000001717 pathogenic effect Effects 0.000 abstract description 2
- 239000012634 fragment Substances 0.000 description 9
- 230000003321 amplification Effects 0.000 description 8
- 238000003199 nucleic acid amplification method Methods 0.000 description 8
- 238000000034 method Methods 0.000 description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 6
- 230000035945 sensitivity Effects 0.000 description 6
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 5
- 238000003745 diagnosis Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 229960002685 biotin Drugs 0.000 description 3
- 235000020958 biotin Nutrition 0.000 description 3
- 239000011616 biotin Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 241000607528 Aeromonas hydrophila Species 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241001509299 Brucella canis Species 0.000 description 2
- 241001148111 Brucella suis Species 0.000 description 2
- 230000004544 DNA amplification Effects 0.000 description 2
- 241000531795 Salmonella enterica subsp. enterica serovar Paratyphi A Species 0.000 description 2
- 241000293869 Salmonella enterica subsp. enterica serovar Typhimurium Species 0.000 description 2
- 241000607764 Shigella dysenteriae Species 0.000 description 2
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 2
- 241000607447 Yersinia enterocolitica Species 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000003317 immunochromatography Methods 0.000 description 2
- 238000011901 isothermal amplification Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 229940007046 shigella dysenteriae Drugs 0.000 description 2
- 229940098232 yersinia enterocolitica Drugs 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000596757 Brucella melitensis M5 Species 0.000 description 1
- 241000274793 Brucella melitensis bv. 1 str. 16M Species 0.000 description 1
- 241001333951 Escherichia coli O157 Species 0.000 description 1
- 241001646719 Escherichia coli O157:H7 Species 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
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- 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
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- C12Q1/6844—Nucleic acid amplification reactions
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Abstract
nucleic acid test paper strip for distinguishing Brucella vaccine strain S19 from natural infection strain relates to the field of brucellosis detection, and solves the problem that the existing pathogen detection method can not detect Brucella vaccine strain S19 and natural infection strain at the same time.
Description
Technical Field
The invention relates to the technical field of brucellosis detection, in particular to a duplex detection card of brucella vaccine strains S19 and natural infection strains and a preparation method and a use method thereof.
Technical Field
In recent years, the epidemic situation of brucella in China has a strong threat, and the nation issues long-term plan of brucella disease in China (2016-2020), wherein quarantine and related prevention and control plans of brucella are specified, which is sufficient for the important role of the brucella disease in prevention and control of the epidemic disease in China.
At present, brucellosis of economic animals such as cattle, sheep and the like is mainly prevented, controlled and radically treated by methods such as vaccine immunization, quarantine clearing and the like, S19 strains (brucella bovis) and S2 strains (brucella suis) are mainly applied to the epidemic prevention of brucellosis at present, wherein S19 strains are mainly used for bovine inoculation, the vaccine strains can infect people and cause abortion of female animals, and the vaccine strains have no immune protection effect on brucella infection of other animals.
In recent years, series of novel molecular diagnosis technologies based on hybridization, amplification and sequencing are continuously emerging, which not only shortens the detection period and reduces the detection cost, but also greatly improves the accuracy and sensitivity of detection and diagnosis of pathogenic microorganisms, but no single molecular diagnosis technologies can simultaneously meet the requirements of simplicity, rapidness, high sensitivity, high accuracy, convenience, practicality and the like.A Chinese patent with the publication number of 105018628A discloses a kit for identifying brucella A19 vaccine strains and wild strains, which is mainly identified by a sequencing-based method, and the specific method is to use PCR amplification-electrophoresis detection to distinguish brucella from other conventional bacterial strains, carry out -step sequencing analysis on PCR amplification products, and identify and diagnose brucella A19 vaccine strains in clinical samples by sequencing results.
Disclosure of Invention
In order to solve the problem that the existing pathogen detection method can not simultaneously detect the Brucella vaccine strain S19 and the naturally infected strain, the invention provides nucleic acid detection test strips for distinguishing the Brucella vaccine strain S19 and the naturally infected strain.
The technical scheme adopted by the invention for solving the technical problem is as follows:
the invention relates to nucleic acid detection test paper strips for distinguishing Brucella vaccine strains S19 from naturally infected strains, which mainly comprise:
the sequence of the forward primer of the Brucella is shown as SEQ ID NO. 1, the sequence of the reverse primer of the Brucella is shown as SEQ ID NO. 2, and the sequence of the probe of the Brucella is shown as SEQ ID NO. 3;
the sequence of the forward primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 4, the sequence of the reverse primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 5, and the sequence of the probe of the Brucella vaccine strain S19 is shown as SEQ ID NO. 6.
The invention relates to nucleic acid detection test paper strips for distinguishing Brucella vaccine strains S19 from naturally infected strains, which mainly comprise:
the sequence of the forward primer of the Brucella is shown as SEQ ID NO. 1, the sequence of the reverse primer of the Brucella is shown as SEQ ID NO. 2, and the sequence of the probe of the Brucella is shown as SEQ ID NO. 3;
the sequence of the forward primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 4, the sequence of the reverse primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 7, and the sequence of the probe of the Brucella vaccine strain S19 is shown as SEQ ID NO. 8.
The invention has the beneficial effects that:
the invention combines the differential segment described in the latest brucellosis diagnosis technology (GBT 18646-.
The RPA-LFD test strip duplex test strips capable of simultaneously detecting Brucella vaccine strain S19 and a naturally infected strain are established, nucleic acid and gene amplification is carried out by adopting fluorescent groups such as FAM, FITC, DIG and the like and/or biotin and other labeled probes and/or primers or labeled primers based on RPA or a related isothermal amplification method, and nucleic acid detection of the Brucella vaccine strain S19 and the naturally infected strain RPA-LFD test strip is carried out by combining with an immunochromatography test strip.
Drawings
FIG. 1 shows the result of the RPA-LFD detection. In the figure, 1, negative control; 2. brucella S19; 3. brucella M5; 4. brucella 16M; 5. brucella S2; 6. brucella RM 6/66.
FIG. 2 shows the result of the specific detection of RPA-LFD, wherein 1 is a negative control, 2 is Brucella S19, 3 is Vibrio parahaemolyticus, 4 is Shigella dysenteriae, 5 is Pseudomonas hydrophila, 6 is Yersinia enterocolitica, 7 is Salmonella typhimurium , 8 is Escherichia coli O157, H7, 9 is Salmonella paratyphi A .
FIG. 3 shows the results of the sensitivity test of Brucella samples. In the figure, 1, blank control; 2. 4.6X 10-1ng/μL；3、4.6×10-2ng/μL；4、4.6×10-3ng/μL；5、4.6×10-4ng/μL；6、4.6×10-5ng/μL。
FIG. 4 shows the results of the sensitivity test of Brucella samples. In the figure, 1, blank control; 2. 4.6X 10-1ng/μL；3、4.6×10-2ng/μL；4、4.6×10-3ng/μL；5、4.6×10-4ng/μL。
Detailed Description
The nucleic acid detection test paper for distinguishing the Brucella vaccine strain S19 from the naturally infected strain is mainly characterized in that biotin and/or a fluorescent group is used for marking a primer and/or a probe, and a chromatographic test paper with two marks is used for quickly detecting a nucleic acid amplification product, wherein the nucleic acid amplification product is obtained by amplifying specific fragments of the Brucella vaccine strain S19 and the naturally infected strain.
The invention relates to nucleic acid detection test paper strips for distinguishing Brucella vaccine strains S19 from naturally infected strains, which mainly comprise:
brucella forward primer sequence F1: catccttacgcgcaacgatatggatcgtttcc are provided.
Brucella reverse primer sequence R1: biotin-tttccggcgcaggcgaaaagggcgtgccgggtctc.
Brucella Probe sequence Probe 1: FAM (or FITC) -accttgcccttgccatcataaaggccggtgc(dSpacer) gttataggcccaataggc-P.
Forward primer sequence F2 of brucella vaccine strain S19: cccttggcattgaacgcggctgtgaaaggacgatg are provided.
Brucella vaccine strain S19 reverse primer sequence R2: biotin-tttccggcgcaggcgaaaagggcgtgccgggtctc.
Brucella vaccine strain S19 Probe sequence Probe 2: DIG-cccttgtccatcaggctttgcttgatatggatga(dispacer) gggcgagactttcggc-P.
Or, the nucleic acid test paper strip for distinguishing the Brucella vaccine strain S19 from the naturally infected strain mainly comprises:
brucella forward primer sequence F1: catccttacgcgcaacgatatggatcgtttcc are provided.
Brucella reverse primer sequence R1: biotin-tttccggcgcaggcgaaaagggcgtgccgggtctc.
Brucella Probe sequence Probe 1: FAM (or FITC) -accttgcccttgccatcataaaggccggtgc(dSpacer) gttataggcccaataggc-P.
Forward primer sequence F2 of brucella vaccine strain S19: cccttggcattgaacgcggctgtgaaaggacgatg are provided.
Brucella vaccine strain S19 reverse primer sequence R2: DIG-tttccggcgcaggcgaaaagggcgtgccgggtctc.
Brucella vaccine strain S19 Probe sequence Probe 2: FAM (or FITC) -cccttgtccatcaggctttgcttgatatggatga(dispacer) gggcgagactttcggc-P.
The primer and the probe are prepared into a test strip with two detection lines according to the existing conventional test strip preparation method, and the test strip is used for identifying the nucleic acid detection of the Brucella vaccine strain S19 and the natural infection strain.
The invention adopts fluorescent groups such as FAM, FITC, DIG and the like, biotin and other labeled probes and primers or labeled primers to carry out nucleic acid and gene amplification based on RPA or related isothermal amplification methods, and combines an immunochromatography test strip to carry out RPA-LFD nucleic acid detection on Brucella vaccine strain S19 and a naturally infected strain. The adopted chromatographic test strip has two detection lines which are respectively marked with FAM or DIG, so that the specific genes of Brucella and the specific genes of Brucella vaccine strain S19 can be identified.
The technical solutions in the embodiments of the present invention will be described clearly and completely below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only partial embodiments of of the present invention, rather than all embodiments.
1. The fluorescence amplification system used in this example was a twist Amp nfo kit from RPA-twist Dx, and the in-system amplification reaction system is shown in the following Table.
The amplification procedure was as follows: mixing the above components, adding into amplification reaction system, adding 2.5 μ L of 280mM MgAc, amplifying at 37-39 deg.C, preferably 37 deg.C, reacting for 20-40min, preferably 20min according to template amount,
the sequence of the Brucella amplified fragment is as follows:
catccttacgcgcaacgatatggatcgtttccgggtaaagcgtcgccagaaggcgcaaatcttccaccttgcccttgccatcataaaggccggtgccgttataggcccaataggcaacgtctgactgcgtaaagccggactccagagcgcccgacttgatcgcattgatattggcaaccgagccattcgacgaaacggccgtcgcgacgagacccgg。
the sequence of the fragment amplified by the Brucella vaccine strain S19 is as follows:
cccttggcattgaacgcggctgtgaaaggacgatgcccgcccttgtccatcaggctttgcttgatatggatgatgggcgagactttcggcacggtgcgggcccatgcgtaaggatcgtagtcgtccgggttagcggatgtcacgtcaccatggtcgatatcggccatcatccacatgggg。
2. after the amplification reaction is finished, diluting the sample according to the volume ratio of 1:50, sucking 10 mu L of diluted reaction solution, dripping the diluted reaction solution into a sample area of a chromatography test strip, then putting the sample area into 100 mu Lbuffer, and reading the result after 15 min.
The results of the detection are shown in FIG. 1. Through research, the nucleic acid detection test strip for distinguishing the Brucella vaccine strain S19 from the naturally infected strain can identify the Brucella and Brucella S19 strains. The invention can amplify specific target amplified fragments and DIG labeled fragments containing FAM labels of Brucella such as Brucella vaccine strain S19, Brucella melitensis M5 and 16M, Brucella suis S2 and Brucella canis RM6/66, and the T2 detection line can amplify specific target fragments containing DIG labels of Brucella such as Brucella S2, Brucella canis RM 6/66.
3. As shown in FIG. 2, it was found from the test results that the test strip of the present invention failed to amplify the FAM-labeled fragment (T1 detection line) and DIG-labeled fragment (T2 detection line) of bacteria such as Vibrio parahaemolyticus, Shigella dysenteriae, Pseudomonas hydrophila, Yersinia enterocolitica, Salmonella typhimurium , Salmonella paratyphi , Escherichia coli O157: H7, etc., whereas Brucella vaccine strain S19 failed to amplify the FAM-labeled fragment (T1 detection line) and T2 detection line.
4. And (3) sensitivity test: as shown in FIG. 3, the nucleic acid of Brucella vaccine strain S19 was diluted by 10X fold and then subjected to RPA-LFD detection, and it was found from the reaction that the detection concentration of Brucella vaccine strain S19 could reach 4.6X 10-3ng/μL。
5. And (3) sensitivity test: as shown in FIG. 4, the nucleic acid sample of Brucella S19 was diluted 10X times with commercially available milk, and then RPA-LFD detection was performed, whereby it was found that the detection concentration of Brucella vaccine strain S19 could reach 4.6X 10-2ng/mu L, the detection effect in milk is obviously lower than that of simple nucleic acid detection, probably because tissue samples such as milk have definite influence on the system.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> military medical institute of military sciences institute of military veterinary research institute
<120> nucleic acid detection test paper strips for distinguishing Brucella vaccine strain S19 from naturally infected strain
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<170>SIPOSequenceListing 1.0
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<213> Brucella forward primer (primer)
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catccttacg cgcaacgata tggatcgttt cc 32
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<213> Brucella reverse primer (primer)
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btntttccgg cgcaggcgaa aagggcgtgc cgggtctc 38
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accttgccct tgccatcata aaggccggtg cdacrgttat aggcccaata ggc 53
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<213> Brucella vaccine strain S19 Forward primer (primer)
<400>4
cccttggcat tgaacgcggc tgtgaaagga cgatg 35
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<213> Brucella vaccine strain S19 reverse primer (primer)
<400>5
btntttccgg cgcaggcgaa aagggcgtgc cgggtctc 38
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<212>DNA
<213> Brucella vaccine strain S19 Probe (primer)
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cccttgtcca tcaggctttg cttgatatgg atgadsacrg ggcgagactt tcggc 55
<210>7
<211>35
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<213> Brucella vaccine strain S19 reverse primer (primer)
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tttccggcgc aggcgaaaag ggcgtgccgg gtctc 35
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<212>DNA
<213> Brucella vaccine strain S19 Probe (primer)
<400>8
cccttgtcca tcaggctttg cttgatatgg atgadsacrg ggcgagactt tcggc 55
Claims (2)
1, kinds of nucleic acid detection test paper strip for distinguishing Brucella vaccine strain S19 and natural infection strain, which is characterized in that it includes:
the sequence of the forward primer of the Brucella is shown as SEQ ID NO. 1, the sequence of the reverse primer of the Brucella is shown as SEQ ID NO. 2, and the sequence of the probe of the Brucella is shown as SEQ ID NO. 3;
the sequence of the forward primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 4, the sequence of the reverse primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 5, and the sequence of the probe of the Brucella vaccine strain S19 is shown as SEQ ID NO. 6.
2, kinds of nucleic acid detection test paper strip for distinguishing Brucella vaccine strain S19 and natural infection strain, which is characterized in that it includes:
the sequence of the forward primer of the Brucella is shown as SEQ ID NO. 1, the sequence of the reverse primer of the Brucella is shown as SEQ ID NO. 2, and the sequence of the probe of the Brucella is shown as SEQ ID NO. 3;
the sequence of the forward primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 4, the sequence of the reverse primer of the Brucella vaccine strain S19 is shown as SEQ ID NO. 7, and the sequence of the probe of the Brucella vaccine strain S19 is shown as SEQ ID NO. 8.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111781378A (en) * | 2020-07-13 | 2020-10-16 | 祁寒松 | Colloidal gold test strip for pathogen and antibody detection and differential diagnosis thereof |
| CN112852698A (en) * | 2021-01-30 | 2021-05-28 | 军事科学院军事医学研究院军事兽医研究所 | Construction method and application of asd gene deletion strain of Brucella A19 strain |
| NL2031171B1 (en) * | 2021-03-19 | 2022-08-12 | Inst Animal Health Guangdong Academy Agricultural Sciences | Primer, Probe and Application for Identifying Brucella Vaccine Strain A 19 and Wild Strain |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950759A (en) * | 2016-06-23 | 2016-09-21 | 中国检验检疫科学研究院 | Kit for simultaneously detecting four pathogenic bacteria and non-diagnostic detection method thereof |
| CN108034739A (en) * | 2018-01-29 | 2018-05-15 | 中国农业科学院兰州兽医研究所 | Differentiate primer, kit and its application of brucella gene-deleted vaccine and natural infection strain based on RPA methods |
| CN108048586A (en) * | 2017-12-22 | 2018-05-18 | 中国动物卫生与流行病学中心 | A kind of detection method of ox kind Brucella sp attenuated vaccine strain S19 |
| CN109439727A (en) * | 2018-04-25 | 2019-03-08 | 内蒙古华星康为生物科技有限公司 | A kind of method and kit detecting microorganism |
-
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- 2019-12-04 CN CN201911225968.3A patent/CN110734993A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105950759A (en) * | 2016-06-23 | 2016-09-21 | 中国检验检疫科学研究院 | Kit for simultaneously detecting four pathogenic bacteria and non-diagnostic detection method thereof |
| CN108048586A (en) * | 2017-12-22 | 2018-05-18 | 中国动物卫生与流行病学中心 | A kind of detection method of ox kind Brucella sp attenuated vaccine strain S19 |
| CN108034739A (en) * | 2018-01-29 | 2018-05-15 | 中国农业科学院兰州兽医研究所 | Differentiate primer, kit and its application of brucella gene-deleted vaccine and natural infection strain based on RPA methods |
| CN109439727A (en) * | 2018-04-25 | 2019-03-08 | 内蒙古华星康为生物科技有限公司 | A kind of method and kit detecting microorganism |
Non-Patent Citations (1)
| Title |
|---|
| LIDE QIN等: "A novel approach for detection of brucella using a real-time recombinase", 《MOLECULAR AND CELLULAR PROBES》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111781378A (en) * | 2020-07-13 | 2020-10-16 | 祁寒松 | Colloidal gold test strip for pathogen and antibody detection and differential diagnosis thereof |
| CN111781378B (en) * | 2020-07-13 | 2023-04-14 | 祁寒松 | Colloidal gold test strip for pathogen and antibody detection and differential diagnosis thereof |
| CN112852698A (en) * | 2021-01-30 | 2021-05-28 | 军事科学院军事医学研究院军事兽医研究所 | Construction method and application of asd gene deletion strain of Brucella A19 strain |
| CN112852698B (en) * | 2021-01-30 | 2022-11-29 | 军事科学院军事医学研究院军事兽医研究所 | Construction method and application of Brucella A19 strain asd gene deletion strain |
| NL2031171B1 (en) * | 2021-03-19 | 2022-08-12 | Inst Animal Health Guangdong Academy Agricultural Sciences | Primer, Probe and Application for Identifying Brucella Vaccine Strain A 19 and Wild Strain |
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