CN104073450A - Goat source arcanobacterium pyogenes and application thereof - Google Patents
Goat source arcanobacterium pyogenes and application thereof Download PDFInfo
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- CN104073450A CN104073450A CN201410072433.8A CN201410072433A CN104073450A CN 104073450 A CN104073450 A CN 104073450A CN 201410072433 A CN201410072433 A CN 201410072433A CN 104073450 A CN104073450 A CN 104073450A
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Abstract
The invention relates to the field of microorganisms and particularly relates to goat source arcanobacterium pyogenes and an application thereof. The biological preservation number of the goat source arcanobacterium pyogenes is CCTCC (China Center For Type Culture Collection) NO: M2014037. The goat source arcanobacterium pyogenes, a specific fragment for DNA molecular identification of the goat source arcanobacterium pyogenes, a pol hemolysin genetic fragment, a recombinant vector and a recombinant strain can be widely applied to researching, diagnosing and treating goat diseased induced by the goat source arcanobacterium pyogenes, thereby laying a foundation for further researching the physiological-biochemical characteristics of the goat source arcanobacterium pyogenes and the damage of the goat source arcanobacterium pyogenes to animal bodies. The invention particularly relates to a preparation method of the specific fragment for DNA molecular identification of the goat source arcanobacterium pyogenes. The preparation method is simple to operate and feasible.
Description
Technical field
The present invention relates to microorganism field, be specifically related to a kind of goat source suppurate concealed bacillus and application thereof.
Background technology
Suppuration Arcanobacterium is in Arcanobacterium, former title corynebacterium pyogenes (Corynebacterium pyogenes), Actinomyces pyogenes (Actinomyces pyogenes), within 1997, name as the concealed bacillus of suppurating (Arcanobacterium pyogenes), a kind of polymorphism, Gram-positive bacillus without mobility, Back ground Information is referring to Baidupedia http://baike.baidu.com/view/9523741.htm.
The concealed bacillus of suppurating is a kind of to ruminant domestic animal and the very important symbiosis conditionality of pig pathogenic bacterium.When animal is in the time that stress situation stimulates as transport, cold, wean and other exception conditions, cause Abwehrkraft des Koepers to decline, the concealed bacillus of suppuration in body or the concealed bacillus of suppuration of having located in the situation of inapparent infection breed in a large number, strengthen pathogenic effects and cause morbidity or worsen.This bacterium infects the pyogenic infection that often causes skin, joint and organ; When serious, often because pyaemia septica is dead.The concealed bacillus that confirmed to suppurate is one of important bacterial pathogen of cattle respiratory disease syndrome; And it is also less to the pathogenic effects research of goat for this bacterium at present.
The concealed bacillus virulence factor of suppuration of having found at present has the concealed bacillus hemolysin pyoysin of suppuration (PLO), collagen binding protein (collagen-binding protein, CbpA), neuraminidase (neuraminidases, Nan), pili fimbriae) etc.These virulence factors, suppurate concealed bacillus in different hosts body perch and cause various courses of infection in play an important role.Chongqing region was introduced good sheep kind in recent years in a large number, and sheep husbandry development rapidly; And breeding production condition is very different, additional southwest is moist sombre winter, causes goat pneumonia to happen occasionally.The sheep that have obstinate quite a lot in scale sheep field, suffers from pneumonia only, mostly poor growth, become thin.
The present invention is directed to the problems referred to above, the concealed bacillus of the suppuration in goat source is explored.
Summary of the invention
In view of this, the invention provides a kind of goat source suppurate concealed bacillus and application thereof, for research, diagnosis and the treatment of the microbial goat disease of concealed bar of suppurating provide new application prospect.
For achieving the above object, technical scheme of the present invention is:
The goat source concealed bacillus of suppurating, biological preserving number is CCTCC NO:M2014037.The described goat source concealed bacillus of suppurating is deposited in Chinese Typical Representative culture collection center on January 22nd, 2014, and address is loujia hill belongs Wuhan University Chinese Typical Representative culture collection center in the school, and deposit number is CCTCC NO:M2014037.The concrete sick sheep from sheep field, Chongqing City, a sheep skeletonize, normal cough, stream nose liquid, the poor growth of separating of this bacterium.The 16S rRNA sequence of this bacterium is carried out to sequence alignment at GenBank, find that institute's calling sequence and suppuration concealed bacillus (strain name: the concealed bacillus of suppurating, logs in numbering: HQ712123) homology are up to 99%; Smear staining microscopy, visible Gram-positive coryneform bacteria, singly or Cheng Cong, exist without obvious chain; The called after concealed bacillus FLZ-0211 (Arcanobacterium pyogenes FLZ-0211) that suppurates.
Further, the described goat source concealed bacillus of suppurating is to separate and obtain from the suppurative necrosis region of the organ such as painstaking effort or lungs of goat pneumonia case.Described separating step comprises: gathers pathological material of disease from the suppurative necrosis region of the organ such as painstaking effort or lungs, asepticly gets pathological material of disease streak inoculation in blood nutrient agar, and 37 DEG C of constant temperature culture 48-72h, separation obtains; Preferably containing 50ml/L blood nutrient agar.
The invention provides a kind of pol hemolysin gene fragment, biological preserving number is the suppurate pol hemolysin gene fragment of concealed bacillus of the goat source of CCTCC M2014037, and sequence is as shown in SEQ ID NO:1.Hemolysin is a kind of extracellular toxin of the concealed bacillus of suppuration, has and dissolves the red corpuscle of many animals, the characteristic of immunocyte, can induce macrophage apoptosis, and this may be relevant to this bacterium infection initiation suppuration focus.Confirm by experiment, be different from other virulence factors, this toxin is ubiquity in concealed bacillus strain isolated is suppurated in goat source.Therefore, the gene of hemolysin, the complementary sequence fragment of pol hemolysin gene fragment or this fragment can be widely used in the detection of the concealed bacillus of suppurating, as the target as detecting the concealed bacillus of suppurating.
The invention provides the suppurate specific fragment of concealed bacillus DNA Molecular Identification of a kind of goat source, biological preserving number is the suppurate 16S rRNA of concealed bacillus of the goat source of CCTCC M2014037, and sequence is as shown in SEQ ID NO:2.The suppurate complementary sequence fragment of specific fragment or specific fragment of concealed bacillus DNA Molecular Identification of this goat source, can be widely used in goat source suppurates in the detection of concealed bacillus, as utilizing this specific fragment, preparation detects the suppurate test kit of concealed bacillus of goat source.
The present invention also provides the preparation method of the specific fragment that a kind of goat source suppurates concealed bacillus DNA Molecular Identification, and the method comprises the following steps:
A, get the above-mentioned goat source concealed bacillus of suppurating, extract genomic dna, prepare template DNA;
B, described masterplate DNA adopt gene universal primer to carry out pcr amplification, separate to obtain the suppurate specific fragment of concealed bacillus DNA Molecular Identification of goat source; Described gene primer is the Auele Specific Primer of the one or more genes in 16S rRNA, pol, nanH, nanP, cbpA, fimA, fimC, fimE and fimG.
Further, described Auele Specific Primer is specially: the Auele Specific Primer of 16S rRNA gene: its upstream primer is as shown in SEQ ID NO:3, and downstream primer is as SEQ ID NO:4; The Auele Specific Primer of pol gene: its upstream primer is as shown in SEQ ID NO:5, and downstream primer is as SEQ ID NO:6; The Auele Specific Primer of nanH gene: its upstream primer is as shown in SEQ ID NO:7, and downstream primer is as SEQ ID NO:8; The Auele Specific Primer of nanP gene: its upstream primer is as shown in SEQ ID NO:9, and downstream primer is as SEQ ID NO:10; The Auele Specific Primer of cbpA gene: its upstream primer is as shown in SEQ ID NO:11, and downstream primer is as SEQ ID NO:12; The Auele Specific Primer of fimA gene: its upstream primer is as shown in SEQ ID NO:13, and downstream primer is as SEQ ID NO:14; The Auele Specific Primer of fimC gene: its upstream primer is as shown in SEQ ID NO:15, and downstream primer is as SEQ ID NO:16; The Auele Specific Primer of fimE gene: its upstream primer is as shown in SEQ ID NO:17, and downstream primer is as SEQ ID NO:18; The Auele Specific Primer of fimG gene:
Its upstream primer is as shown in SEQ ID NO:19, and downstream primer is as SEQ ID NO:20.
Described pcr amplification reaction carries out according to a conventional method.Described separation can be reclaimed PCR product the specific fragment of DNA molecular qualification after 1% agarose gel electrophoresis, be cloned into pMD18-T carrier, Transformed E .coli DH5 α bacterial strain, screening positive clone, and use carrier primer M13+/M13-to carry out two-way order-checking.16S rRNA gene order in the 16S rRNA gene order of mensuration and GenBank is carried out to homology comparison, from goat suppuration focus, be separated to the concealed bacillus of suppurating.
On the basis of above-mentioned research, the invention provides a kind of recombinant vectors, the suppurate specific fragment of concealed bacillus DNA Molecular Identification of the goat source by sequence as shown in SEQ ID NO:2 is cloned into pMD18-T carrier and forms.Further, the present invention also provides a kind of recombinant bacterial strain, is converted into E.coli DH5 α bacterial strain forms by described recombinant vectors.This recombinant vectors and recombinant bacterial strain can be widely used in goat source and suppurate in the application of research and product etc. of concealed bacillus.
Useful technique effect of the present invention is:
Concealed bacillus, goat source suppurate specific fragment, pol hemolysin gene fragment, recombinant vectors and the recombinant bacterial strain of concealed bacillus DNA Molecular Identification suppurate in goat of the present invention source, can be widely used in research, diagnosis and the treatment of the concealed bar microbial goat disease of suppurating, be suppurate concealed bacillus physio-biochemical characteristics and the harm of animal body is laid a good foundation of further research goat source.The suppurate preparation method of specific fragment of concealed bacillus DNA Molecular Identification of goat of the present invention source is simple to operate, feasible.
Brief description of the drawings
Fig. 1 is that the suppurate PCR of concealed bacillus virulence factor of goat source identifies (wherein, M is that the gene fragment length that DNA Marker DL2000(M band shows to top successively from picture bottom is: 100bp, 250bp, 500bp, 750bp, 1000bp, 2000bp), 1 is pol, 2 is nanH, and 3 is nanP, and 4 is cbpA, 5 is fimA, 6 is fimC, and 7 is fimE, and 8 is fimG);
Fig. 2 is the PCR qualification (wherein, M is DNA Marker DL2000, and 9 is the object band that the concealed bacillus 16S rRNA gene of suppuration is about 1500bp) of the concealed bacillus part 16S rRNA gene of suppuration.
Embodiment
Hereinafter with reference to accompanying drawing, embodiments of the present invention is described in detail.The experimental technique of unreceipted actual conditions in preferred embodiment, carries out according to normal condition.For example, condition described in molecular cloning experiment guide (Huang Peitang etc. translate, Science Press, 2002 for the third edition, the work such as J. Pehanorm Brooker), or the condition of advising according to manufacturer is carried out.The test materials that the present invention is used, if no special instructions, is commercially available purchase product.
1. pathological material of disease: pathological material of disease is from the sick sheep in sheep field, Chongqing City, and a sheep skeletonize, normal cough, stream nose liquid, poor growth, gather the suppurative necrosis regions of organ such as disease sheep painstaking effort and lungs.
2. bacterial strain and main agents: E.coli DH5 α, is preserved by Chongqing Academy of Animal Sciences's veterinary institute; RTaq archaeal dna polymerase, pMDl8-T carrier, dNTPs and DNA Marker, DNA glue reclaim test kit, purchased from precious biotechnology (Dalian) company limited; Genome DNA extracting reagent kit, purchased from TIANGEN company.
3. laboratory animal: clean level kunming mice 3 week age, purchased from Chongqing Teng Xin Bioisystech Co., Ltd; 1 the monthly age goat, purchased near goat plant.
4. the preparation of substratum
Nutrient agar, purchased from Hangzhou microorganism reagent company limited;
Blood nutrient agar, is prepared by Chongqing Academy of Animal Sciences's veterinary institute.Method is as follows: sterilizing adds the triangular flask of granulated glass sphere, through jugular vein aseptic collection goat blood, and at the uniform velocity defiber sheep blood is made in concussion, is cooled to the ordinary nutrient agar substratum of 45-50 DEG C after adding sterilizing in 6% ratio, after shaking up gently, be sub-packed in while hot and in plate, make flat board;
5. separate
The pathological material of disease of aseptic collection is inoculated in containing 50ml/L blood nutrient agar, cultivates 48h for 37 DEG C and observe colony growth characteristic.Get single colony inoculation in cultivating containing 50ml/L blood nutrient agar is dull and stereotyped, pure growth is used for extracting genomic dna, extracts by genome DNA extracting reagent kit specification sheets.Carry out according to a conventional method PCR reaction.Primer is synthesized by Sangon Biotech (Shanghai) Co., Ltd., and primer sequence is in table 1.
The primer of the concealed bacillus of table 1PCR amplification suppuration
6. qualification
1) PCR of virulence factor qualification:
PCR product, through 1% agarose gel electrophoresis, detects Major Virulence Factors hemolysin gene plo, Neuraminidase Gene nanH and nanP, collagen binding protein cbpA, fimbriae gene fimA, fimC, fimE and the fimG of the concealed bacillus of suppurating.As shown in Figure 1, wherein, M is DNA Marker DL2000 to its electrophorogram, and 1 is pol, and 2 is nanH, and 3 is nanP, and 4 is cbpA, and 5 is fimA, and 6 is fimC, and 7 is fimE, and 8 is fimG.Qualification result shows, the concealed bacillus strain isolated that suppurates exists hemolysin gene plo, Neuraminidase Gene nanH and nanP, collagen binding protein gene cbpA, fimbriae gene fimC and fimE, without fimbriae gene fimA and fimG.
The concealed bacillus of known suppuration has host infection scope and infected tissue widely, and to contain the virulence factor such as collagen binding protein and neuraminidase relevant with it for this.Collagen generally distributes in higher animal tissue, for utilizing collagen binding protein to adhere to infected tissue, the concealed bacillus of suppurating provides possibility, and neuraminidase can decompose the sialic acid of host cell, and reduce the glutinousness of film surface mucus, be convenient to bacterium and slip into deep tissues.In the two field planting at thalline and invasive procedure, play an important role.To the Molecular Detection result of the concealed bacillus virulence factor that suppurates as Fig. 1, this result shows that this strain isolated contains hemolysin gene plo, Neuraminidase Gene nanH and nanP, collagen binding protein gene cbpA, fimbriae gene fimC and fimC simultaneously, points out its virulence may be stronger.
In addition, from isolate, amplify the 270bp special hemolysin gene pol fragment of concealed bacillus of suppurating.Hemolysin gene pol fragment produces hemolysin, hemolysin is a kind of extracellular toxin of the concealed bacillus of suppuration, have and dissolve the red corpuscle of many animals, the characteristic of immunocyte, can induce macrophage apoptosis, this may be relevant to this bacterium infection initiation suppuration focus, be different from other virulence factors, this toxin is at the ubiquity in concealed bacillus strain isolated that suppurates.Therefore, available its gene is as the target that detects the concealed bacillus of suppurating.
2) the goat source qualification of specific fragment of concealed bacillus DNA Molecular Identification of suppurating:
Utilize bacterium universal primer to carry out PCR qualification to the concealed bacillus of suppurating, result, as Fig. 2, amplifies the object band that is about 1500bp.The specific fragment of the DNA molecular qualification that PCR product reclaims after 1% agarose gel electrophoresis, be cloned into pMD18-T carrier, Transformed E .coli DH5 α bacterial strain, screening positive clone uses carrier primer M13+/M13-to carry out two-way order-checking, and the 16S rRNA gene order in the 16S rRNA gene order of mensuration and GenBank is carried out to homology comparison.
Draw by bacterial 16 S rRNA gene identification, from goat suppuration focus, separate the concealed bacillus that obtains suppurating.Smear staining microscopy, visible Gram-positive coryneform bacteria, singly or Cheng Cong, exist without obvious chain.7. challenge test
The single bacterium colony of picking, is inoculated in containing 50ml/L blood broth culture, and 48h is cultivated in 37 DEG C of concussions, then carries out live bacterial count, adjusts bacterial concentration to 1 × 10
9cFU/ml, 0.2ml/ mouse of abdominal injection bacterium liquid.Observe clinical symptom, dead mouse is weighed, cuts open inspection, bacterium separation and Culture and cause of disease qualification, not dead mouse is carried out to weighing body weight, after execution, cut open inspection.
Mouse all shows the phenomenons such as spirit is depressed, appetite stimulator after concealed bacillus is suppurated in inoculation after about 1h, after 1d, symptom takes a turn for the better to some extent.After 7d, all mouse are not dead, put to death mouse.Approximately 1/3 mouse gets well, and has dermapostasis at inoculation position, and the speed of growth and normal mouse are suitable.All the other mouse stop growing, and become thin, and body weight declines before attacking poison, and the mental status is not good.Cuing open the inspection mouse organ of not finding to get well has obviously visible pathology, except dermapostasis.The mouse that becomes thin has serious peritonitis, purulence peritonaeum parcel liver, and liver spreads all over blutpunkte, has a large amount of necrotic lesions, stomach, the serious oedema of enteron aisle, mucous membrane comes off, empty without food.Other organ-tissues are without obvious pathology.From above-mentioned mouse focus, separate, identify the concealed bacillus of suppurating.
Goat challenge test shows that pathology mainly concentrates on inoculation position.These result lesions showed occur in inoculation position or next-door neighbour's inoculation position, and prompting cause of disease is difficult for diffusion in vivo.But pathogenic bacteria looks infection site and tissue depth to the harm of animal and is different.From clinical case and challenge test result, the goat concealed coli infections that suppurates is that chronic expendable infects, and acute death can not occur.But this chronic infection is polyinfection has created condition, disseminates for a long time cause of disease, makes epidemic situation complicated.
Finally explanation is, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the art is to be understood that, can modify or be equal to replacement technical scheme of the present invention, and not departing from aim and the scope of technical solution of the present invention, it all should be encompassed in the middle of claim scope of the present invention.
Claims (10)
1. the goat source concealed bacillus of suppurating, is characterized in that: biological preserving number is CCTCC NO:M2014037.
2. the goat according to claim 1 source concealed bacillus of suppurating, is characterized in that: the described goat source concealed bacillus of suppurating is to separate and obtain from the painstaking effort of goat pneumonia case or the suppurative necrosis region of lungs.
3. biological preserving number is the suppurate pol hemolysin gene fragment of concealed bacillus of the goat source of CCTCC NO:M2014037, and sequence is as shown in SEQ ID NO:1.
4. the application that described in claim 3, the complementary sequence fragment of pol hemolysin gene fragment or this fragment is suppurated in concealed bacillus in detection.
5. the goat source specific fragment of concealed bacillus DNA Molecular Identification that suppurates, sequence is as shown in SEQ ID NO:2.
6. the goat claimed in claim 5 source preparation method of specific fragment of concealed bacillus DNA Molecular Identification of suppurating, is characterized in that, comprises the following steps:
A, the goat source described in claim 1 or 2 of the getting concealed bacillus of suppurating, extracts genomic dna, prepares template DNA;
B, described masterplate DNA adopt gene universal primer to carry out pcr amplification, separate to obtain the suppurate specific fragment of concealed bacillus DNA Molecular Identification of goat source; Described gene primer is the Auele Specific Primer of the one or more genes in 16S rRNA, pol, nanH, nanP, cbpA, fimA, fimC, fimE and fimG.
7. preparation method according to claim 6, is characterized in that: described Auele Specific Primer is specially:
The Auele Specific Primer of 16S rRNA gene: its upstream primer is as shown in SEQ ID NO:3, and downstream primer is as SEQ ID NO:4;
The Auele Specific Primer of pol gene: its upstream primer is as shown in SEQ ID NO:5, and downstream primer is as SEQ ID NO:6;
The Auele Specific Primer of nanH gene: its upstream primer is as shown in SEQ ID NO:7, and downstream primer is as SEQ ID NO:8;
The Auele Specific Primer of nanP gene: its upstream primer is as shown in SEQ ID NO:9, and downstream primer is as SEQ ID NO:10;
The Auele Specific Primer of cbpA gene: its upstream primer is as shown in SEQ ID NO:11, and downstream primer is as SEQ ID NO:12;
The Auele Specific Primer of fimA gene: its upstream primer is as shown in SEQ ID NO:13, and downstream primer is as SEQ ID NO:14;
The Auele Specific Primer of fimC gene: its upstream primer is as shown in SEQ ID NO:15, and downstream primer is as SEQ ID NO:16;
The Auele Specific Primer of fimE gene: its upstream primer is as shown in SEQ ID NO:17, and downstream primer is as SEQ ID NO:18;
The Auele Specific Primer of fimG gene: its upstream primer is as shown in SEQ ID NO:19, and downstream primer is as SEQ ID NO:20.
8. the goat claimed in claim 5 source application that the specific fragment of concealed bacillus DNA Molecular Identification or the complementary sequence fragment of this fragment are detecting goat source and suppurate in concealed bacillus of suppurating.
9. a recombinant vectors, is characterized in that: be cloned into pMD18-T carrier and formed by the suppurate specific fragment of concealed bacillus DNA Molecular Identification of goat claimed in claim 5 source.
10. a recombinant bacterial strain, is characterized in that: be converted into E.coli DH5 α bacterial strain by recombinant vectors claimed in claim 9 and form.
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| CN104561259A (en) * | 2014-10-23 | 2015-04-29 | 中华人民共和国黄埔出入境检验检疫局 | Primers, probe and detection method for real-time fluorescent PCR detection of arcanobacterium pyogenes |
| CN104561259B (en) * | 2014-10-23 | 2017-01-04 | 中华人民共和国黄埔出入境检验检疫局 | The real-time PCR detection primer of a kind of Arcanobacterium pyogenes and probe and detection method |
| CN107466326A (en) * | 2015-03-12 | 2017-12-12 | 硕腾服务有限责任公司 | Hemolysin method and composition |
| CN108048588A (en) * | 2018-01-17 | 2018-05-18 | 重庆市畜牧科学院 | The detection primer and detection kit of a kind of Arcanobacterium pyogenes |
| CN109337996A (en) * | 2018-12-24 | 2019-02-15 | 福建省农业科学院畜牧兽医研究所 | It is a kind of for detecting the primer and probe of goat Arcanobacterium pyogenes |
| CN109439779A (en) * | 2018-12-24 | 2019-03-08 | 林裕胜 | One kind is for detecting goat Arcanobacterium pyogenes real-time fluorescence quantitative PCR primer |
| CN109929780A (en) * | 2019-03-25 | 2019-06-25 | 江西省农业科学院畜牧兽医研究所 | One boar source Arcanobacterium pyogenes and its application |
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| CN112611864A (en) * | 2020-12-03 | 2021-04-06 | 重庆市畜牧科学院 | System and method for screening germ models |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104561259A (en) * | 2014-10-23 | 2015-04-29 | 中华人民共和国黄埔出入境检验检疫局 | Primers, probe and detection method for real-time fluorescent PCR detection of arcanobacterium pyogenes |
| CN104561259B (en) * | 2014-10-23 | 2017-01-04 | 中华人民共和国黄埔出入境检验检疫局 | The real-time PCR detection primer of a kind of Arcanobacterium pyogenes and probe and detection method |
| CN107466326A (en) * | 2015-03-12 | 2017-12-12 | 硕腾服务有限责任公司 | Hemolysin method and composition |
| CN107466326B (en) * | 2015-03-12 | 2021-10-15 | 硕腾服务有限责任公司 | Hemolysin methods and compositions |
| CN108048588B (en) * | 2018-01-17 | 2021-04-06 | 重庆市畜牧科学院 | Detection primer and detection kit for cryptococcus pyogenes |
| CN108048588A (en) * | 2018-01-17 | 2018-05-18 | 重庆市畜牧科学院 | The detection primer and detection kit of a kind of Arcanobacterium pyogenes |
| CN109439779A (en) * | 2018-12-24 | 2019-03-08 | 林裕胜 | One kind is for detecting goat Arcanobacterium pyogenes real-time fluorescence quantitative PCR primer |
| CN109337996A (en) * | 2018-12-24 | 2019-02-15 | 福建省农业科学院畜牧兽医研究所 | It is a kind of for detecting the primer and probe of goat Arcanobacterium pyogenes |
| CN109929780A (en) * | 2019-03-25 | 2019-06-25 | 江西省农业科学院畜牧兽医研究所 | One boar source Arcanobacterium pyogenes and its application |
| CN109929780B (en) * | 2019-03-25 | 2021-12-17 | 江西省农业科学院畜牧兽医研究所 | Humanized cryptobacter pus and application thereof |
| CN110642927A (en) * | 2019-10-16 | 2020-01-03 | 重庆市畜牧科学院 | Application of protein in preparation of medicine for preventing cryptococcus pyogenes infection |
| CN110642927B (en) * | 2019-10-16 | 2023-04-14 | 重庆市畜牧科学院 | Application of protein in preparation of medicine for preventing cryptococcus pyogenes infection |
| CN112611864A (en) * | 2020-12-03 | 2021-04-06 | 重庆市畜牧科学院 | System and method for screening germ models |
| CN112611864B (en) * | 2020-12-03 | 2024-03-12 | 重庆市畜牧科学院 | System and method for screening germ model |
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