CN103941013A - Rapid detection test strip for actinobacillus pleuropneumoniae endotoxin - Google Patents
Rapid detection test strip for actinobacillus pleuropneumoniae endotoxin Download PDFInfo
- Publication number
- CN103941013A CN103941013A CN201310083779.3A CN201310083779A CN103941013A CN 103941013 A CN103941013 A CN 103941013A CN 201310083779 A CN201310083779 A CN 201310083779A CN 103941013 A CN103941013 A CN 103941013A
- Authority
- CN
- China
- Prior art keywords
- layer
- antibody
- trace
- apx
- monoclonal antibody
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000012360 testing method Methods 0.000 title claims abstract description 75
- 241000606748 Actinobacillus pleuropneumoniae Species 0.000 title claims abstract description 32
- 239000002158 endotoxin Substances 0.000 title claims abstract description 14
- 238000001514 detection method Methods 0.000 title abstract description 26
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 30
- 229920002678 cellulose Polymers 0.000 claims abstract description 24
- 239000001913 cellulose Substances 0.000 claims abstract description 24
- 239000012528 membrane Substances 0.000 claims abstract description 15
- 241000283973 Oryctolagus cuniculus Species 0.000 claims abstract description 12
- 239000000463 material Substances 0.000 claims abstract description 12
- 238000010521 absorption reaction Methods 0.000 claims abstract description 8
- 239000010410 layer Substances 0.000 claims description 44
- 238000002372 labelling Methods 0.000 claims description 31
- 239000010931 gold Substances 0.000 claims description 29
- 229910052737 gold Inorganic materials 0.000 claims description 29
- 238000002360 preparation method Methods 0.000 claims description 27
- 239000000835 fiber Substances 0.000 claims description 15
- 239000011491 glass wool Substances 0.000 claims description 14
- 241001494479 Pecora Species 0.000 claims description 12
- 239000002250 absorbent Substances 0.000 claims description 9
- 230000002745 absorbent Effects 0.000 claims description 9
- 239000002245 particle Substances 0.000 claims description 9
- 108010088160 Staphylococcal Protein A Proteins 0.000 claims description 8
- 239000003550 marker Substances 0.000 claims description 7
- 239000000020 Nitrocellulose Substances 0.000 claims description 6
- 229920001220 nitrocellulos Polymers 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000002033 PVDF binder Substances 0.000 claims description 4
- 239000000123 paper Substances 0.000 claims description 4
- 239000004033 plastic Substances 0.000 claims description 4
- 229920002981 polyvinylidene fluoride Polymers 0.000 claims description 4
- 241000446313 Lamella Species 0.000 claims description 2
- 230000021523 carboxylation Effects 0.000 claims description 2
- 238000006473 carboxylation reaction Methods 0.000 claims description 2
- 230000001681 protective effect Effects 0.000 claims description 2
- 239000011241 protective layer Substances 0.000 claims description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 6
- 244000144972 livestock Species 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 abstract 2
- 238000001179 sorption measurement Methods 0.000 abstract 2
- 241000283707 Capra Species 0.000 abstract 1
- 238000009395 breeding Methods 0.000 abstract 1
- 230000001488 breeding effect Effects 0.000 abstract 1
- 239000012481 endotoxin detection reagent Substances 0.000 abstract 1
- 235000013372 meat Nutrition 0.000 abstract 1
- 238000012856 packing Methods 0.000 abstract 1
- 244000144977 poultry Species 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 20
- 241000282898 Sus scrofa Species 0.000 description 18
- 201000010099 disease Diseases 0.000 description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 18
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 description 14
- 108700012359 toxins Proteins 0.000 description 12
- 238000002965 ELISA Methods 0.000 description 11
- 239000007788 liquid Substances 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 210000002966 serum Anatomy 0.000 description 10
- 239000003053 toxin Substances 0.000 description 10
- 231100000765 toxin Toxicity 0.000 description 10
- 208000015181 infectious disease Diseases 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000003745 diagnosis Methods 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 description 5
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical class N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 5
- 239000000427 antigen Substances 0.000 description 5
- 108091007433 antigens Proteins 0.000 description 5
- 102000036639 antigens Human genes 0.000 description 5
- 230000001717 pathogenic effect Effects 0.000 description 5
- 241000204031 Mycoplasma Species 0.000 description 4
- 206010035664 Pneumonia Diseases 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 244000052769 pathogen Species 0.000 description 4
- 201000006509 pleuropneumonia Diseases 0.000 description 4
- 208000031504 Asymptomatic Infections Diseases 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000010241 blood sampling Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000012797 qualification Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 244000208060 Lawsonia inermis Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 238000002405 diagnostic procedure Methods 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000011888 foil Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000035931 haemagglutination Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 230000000521 hyperimmunizing effect Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 230000001018 virulence Effects 0.000 description 2
- 208000002874 Acne Vulgaris Diseases 0.000 description 1
- 241000606750 Actinobacillus Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 241000726221 Gemma Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 101000723151 Hemachatus haemachatus Short neurotoxin 2 Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 101000740663 Leiurus hebraeus Toxin Lqh4 Proteins 0.000 description 1
- 101000740664 Leiurus quinquestriatus quinquestriatus Alpha-toxin Lqq4 Proteins 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 101710116435 Outer membrane protein Proteins 0.000 description 1
- 241000606860 Pasteurella Species 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 229920001030 Polyethylene Glycol 4000 Polymers 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 238000005411 Van der Waals force Methods 0.000 description 1
- 206010000496 acne Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009739 binding Methods 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 208000030303 breathing problems Diseases 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 231100000284 endotoxic Toxicity 0.000 description 1
- 230000002346 endotoxic effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000012092 latex agglutination test Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 210000003928 nasal cavity Anatomy 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- QQONPFPTGQHPMA-UHFFFAOYSA-N propylene Natural products CC=C QQONPFPTGQHPMA-UHFFFAOYSA-N 0.000 description 1
- -1 propylene glucosan Chemical compound 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000012153 swine disease Diseases 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000005570 vertical transmission Effects 0.000 description 1
- 239000000304 virulence factor Substances 0.000 description 1
- 230000007923 virulence factor Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/30—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Mycoplasmatales, e.g. Pleuropneumonia-like organisms [PPLO]
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Physics & Mathematics (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to an actinobacillus pleuropneumoniae endotoxin detection reagent displaying appliance, and especially relates to a rapid detection test strip for actinobacillus pleuropneumoniae endotoxin (ApxIV). The test strip contains a support layer and a reaction reagent carrier adsorption layer; the support layer is a water-nonabsorbent thin strip, and the reaction reagent carrier adsorption layer is pasted on the support layer; the test strip successively comprises a fibrous layer, an ApxIV gold-labeled monoclonal-antibody or polyclonal-antibody fibrous layer and a cellulose membrane layer from the sample test end, and the handle end is a water-absorption material layer; an ApxIV paired monoclonal antibody or polyclonal antibody or monoclonal antibody solution is sprayed on the fibrous layer for obtaining a detection trace '|', '/' or '\'; and a goat (rabbit) anti-mouse or anti-pig IgG polyclonal antibody or a SPA solution is sprayed on the cellulose membrane layer for obtaining a contrast trace '|', '/' or '\'. The detection test strip is specific, sensitive, intuitionistic, accurate, convenient and rapid, and can be popularized and applied in departments relevant to livestock and poultry breeding, meat packing and quarantine and the like.
Description
one, technical field
The present invention is the detection reagent displaying appliance that one relates to actinobacillus pleuropneumoniae endotoxin (ApxIV), particularly relates to one and can detect the test strip of actinobacillus pleuropneumoniae endotoxin (ApxIV).
two, technical background
Pig pleuropneumonia is the breathing problem of a kind of acute, hot, the hyperinfection of the pig that caused by actinobacillus pleuropneumoniae (APP).Actinobacillus pleuropneumoniae is the member of Pasteurella section Actinobacillus, is the little coccobacillus of a kind of Gram-negative, and being of having is very thin shaft-like or thread, shows as multiform state property and the two poles of the earth coloring.There is pod membrane, have pili, without gemma.Clinically taking acute hemorrhagic fiber disposition pleuropneumonia and chronic fiber disposition gangrenosum acne pleuropneumonia as feature.This disease is distributed in world many countries and area, is a kind of worldwide disease, is at present the fashion trend of rising.China, in this disease of alleged occurrence in 1987, causes great economic loss to large scale of pig farm field.The variation of weather and the arriving of autumn and winter season easily cause the generation of this disease and popular, should attract great attention, and take effective measures, and avoid the generation of this disease cold season and popular.
The pig that carries disease germs of sick pig, subclinical infection is that the sow of the main infection sources, particularly subclinical infection of this disease is the main reservoir host of cause of disease, in the propagation of this bacterium, plays an important role.Although this disease can not infect piglet by vertical transmission, can, by directly contacting through respiratory tract infection, have influence on the safety of piglet.By air borne, can pass to another pig house from a morbidity pig house great-jump-forward.Also can by contact by the vehicle of pathogen contamination, instrument, apparatus and keeper flow etc. infect indirectly.Rodent and birds also can be propagated this disease.The pig of various ages, sex has neurological susceptibility, wherein more with the pig morbidity in 6～16 week age, the highest with the pig incidence of disease at 3 monthly ages, can reach 80%～100%, and mortality ratio is generally 50%.
Pig pleuropneumonia is one of principal disease of current harm countries in the world pig industry.At present APP finds that there is 15 kinds of serotypes altogether, and the popular advantage serotype of every country is different.Between different serotypes and the virulence of the different strains of same serotype all there are differences, pathogenic also have power point, thereby cause this disease diagnosis and treatment very difficult.Research in recent years shows, the factor relevant to pathogen virulence has toxin, pod membrane, outer membrane protein, urase etc., wherein toxin is main virulence factor, also be main immunogene, can be used as diagnostic antigen and set up Serology test, can also be used for diagnosis and weigh immune state, therefore toxin study has become this sick study hotspot.APP has 4 kinds of toxin, i.e. toxin
, II,
and the toxin IV of recent findings, they all belong to RTX (Repeatsinthe structural, RTX) toxin family.In recent years find, do not have the APP of One serotype can secrete 4 kinds of toxin, but all serotype of APP can be secreted ApxlV toxin protein in infection animal body, and the bacterium of in vitro culture can not secrete this toxin simultaneously.Therefore, ApxlV toxin detected in animal body, just can prove the infection of APP, ApxlV toxin protein is being set up larger theory value and the application prospect of tool aspect diagnostic method, can be used for infection and the non-infection of difference diagnosis APP.At present the diagnostic method of above-mentioned disease is mainly contained following several.
(1) microscopy.Get nasal cavity and bronchial secretion or pulmonary lesion material smear, Gram’s staining, microexamination, as see dense gram-negative little coccobacillus or the bacillus exilis dying in polymorphic the two poles of the earth, can do further qualification.
(2) pathogen separation qualification.The aseptic pathological material of disease of getting, intersect streak inoculation with 5% Blood In Sheep agar plate and staphylococcus aureus, under 5%～10% gas concentration lwevel condition, cultivate 24 hours for 37 DEG C, around staphylococcus aureus, have satellite colony, then carry out the biochemical identification such as urase and mannose fermentation.And carry out that CAMP (CAMP) test, hemolytic are measured and the inspection such as serotype.
(3) serodiagnosis.Fluorescent antibody technics can be used for the inspection of specific antigen; Enzyme linked immunosorbent assay (ELISA) can be used for detecting antibody with complement fixation test (CFT), finds the pig of subclinical infection.Indirect hemagglutination test (IHA), the indirect hemagglutination test of APP is for detection and the somatotype of APP.Latex agglutination test is a kind of simple and rapid detection method, and it both can, for the diagnosis of APP, also can be used for somatotype.Carry out the serodiagnosis of APP with enzyme linked immunosorbent assay technology, can be divided into type specificity ELISA and species specificity ELISA.The former can only make detection to the APP of One serotype, and the latter can make detection to the APP of all serotypes.
(4) PCR (PCR) technology: round pcr has the advantage such as high sensitivity, high specific, and multiple PCR method has been widely used in the qualification of APP or ApxIV and epidemiology survey.
Above-mentioned detection method needs professional in laboratory operation, complex operation, and detection is wasted time and energy; And need expensive instrument and equipment, as PCR instrument, microplate reader etc., for layman, above-mentioned detection method has been difficult to.Although the special sensitivity of said method, cannot realize field quick detection or diagnosis.The present invention, research a kind of easy fast, real-time online Test paper, to controlling and to eliminate this disease significant.
three, summary of the invention
The object of the invention is the shortcoming that detects the existence of swine disease cause of disease in prior art in order to overcome, provide a kind of special, responsive, easy fast detecting actinobacillus pleuropneumoniae endotoxic method, develop the test strip that detects actinobacillus pleuropneumoniae endotoxin (ApxIV).
Technical scheme of the present invention is: the test strip that a kind of actinobacillus pleuropneumoniae endotoxin (ApxIV) is provided, this test strips contains supporting layer and adsorbed layer, supporting layer is the lamella not absorbing water, adsorbed layer is attached on supporting layer, adsorbed layer is followed successively by the absorbent material layer of sample adsorbing fiber layer, golden labeling antibody fibrage, cellulose rete and handle end from test lead, be provided with and detect trace and contrast trace on cellulose rete; The absorption of gold labeling antibody fibrage has the monoclonal antibody of the anti-ApxIV of nanometer grade gold particle marker, detects the pairing monoclonal antibody of the anti-ApxIV of trace and prints, the polyclonal antibody of sheep or the anti-mouse IgG of rabbit for contrast trace; Or golden labeling antibody fibrage absorption has the polyclonal antibody of the anti-ApxIV of nanometer grade gold particle marker, the monoclonal antibody preparation of anti-ApxIV for detection trace, contrast staphylococcal protein A (SPA) or how anti-preparation of anti-pig IgG for trace.
Detecting the pairing monoclonal antibody preparation of the anti-ApxIV of trace prepares with the pairing monoclonal antibody solution of ApxIV; The polyclonal antibody preparation that detects the anti-ApxIV of trace is the polyclonal antibody preparation with ApxIV.
Supporting layer is made with the hard plastic slip or the cardboard bar that do not absorb water; Test lead sample adsorbing fiber layer is made with glass wool; Gold labeling antibody fibrage is made with glass wool and golden labeling antibody, and golden labeling antibody can be monoclonal antibody or polyclonal antibody.
Cellulose rete is made with nitrocellulose filter or pure cellulose film or carboxylation cellulose membrane or polyvinylidene fluoride PVDF cellulose membrane.
Absorbent material layer is made with thieving paper.
Detect trace and contrast trace is orthoscopic or oblique line formula, on cellulose rete, contain one and detect trace and one contrast trace, the spread pattern that detects trace and contrast trace be "
||", "
//", "
?" in any.
Above test strips adsorbed layer, contain layer protective layer; protective seam is attached on adsorbed layer; on test lead sample adsorbing fiber layer, golden labeling antibody fibrage and absorbent material layer, be coated with diaphragm; on the test lead sample adsorbing fiber layer diaphragm corresponding with golden labeling antibody fibrage intersection, be printed with sample mark line, this mark line deflection test lead sample adsorbing fiber layer one about 0.5cm place of side place.
As required, select above-mentioned golden labeling antibody fibrage, detect a kind of form in trace and contrast trace spread pattern.
Positive beneficial effect of the present invention:
1. detection specificity is strong, susceptibility is high: test strip of the present invention is made as basis taking nanometer grade gold particle marker high-affinity monoclonal antibody specific or specific polyclonal antibody, in gold labeling antibody, between gold grain and antibody molecule, form without covalent bond, the two combines by the Van der Waals force between the charges of different polarity, gold grain does not affect specificity and the adhesion of monoclonal antibody or polyclonal antibody, and has higher mark rate.Test strip of the present invention has higher specificity and susceptibility, nanogram level actinobacillus pleuropneumoniae endotoxin can be detected.
2. easy and simple to handle, quick: to use when ELISA test strip of the present invention without additional any Other Instruments and reagent, its test lead need be inserted in sample liquid to be checked about 30 seconds, then in 1-5 minute, can judge testing result.
3. testing result is directly perceived, accurate: whether test strips of the present invention is to show that henna detection line and control line are as the foundation of judging positive and negative findings, only show a brownish red control line C at the control line marking place of cellulose membrane, and show without brownish red band at detection line marking place, represent the negative result of detected ApxIV; Control line marking place at cellulose membrane shows a brownish red control line C, detecting a brownish red band T of trace place demonstration, represents the positive result of detected ApxIV.No matter positive findings or negative findings control line C all should show, in the time that control line C does not show, illustrate that test strips lost efficacy.
4. testing cost reduces: use test strip of the present invention, do not need Other Instruments and reagent, saved instrument, equipment and additive reagent expense; Layman is real-time online detection at any time also, without paying expert diagnosis Laboratory Fee and correlative charges thereof, can reduce greatly the input of testing cost, reduces testing cost.
5. usable range is wide: test strip of the present invention is simple to operate, i.e. " foolproof " operation, and easy to carry, easily preservation, can meet the not needs of commensurate and different levels personnel, comprise that professional chemical examination, customs quarantine control, health and epidemic prevention, quality monitoring, livestock products processing, intensive culture arrive individual cultivation etc., have wide market outlook and social benefit.
four, brief description of the drawings:
The side-looking structural representation of the test strip of a kind of actinobacillus pleuropneumoniae endotoxin of Fig. 1 (ApxIV)
The plan structure schematic diagram of the test strip of a kind of actinobacillus pleuropneumoniae endotoxin of Fig. 2 (ApxIV)
five, embodiment:
Following examples only, in order to further illustrate the present invention, do not limit content of the present invention.The preparation of the test strip of actinobacillus pleuropneumoniae endotoxin (ApxIV), need to prepare monoclonal antibody and the polyclonal antibody of anti-ApxIV, for the preparation of detecting trace and golden labeling antibody fibrage; Need to prepare sheep or rabbit anti-mouse igg antibody, the anti-pig IgG antibody of sheep or rabbit, for the preparation of contrast trace simultaneously.
1. the preparation of the anti-mouse of sheep (rabbit) or anti-pig IgG antibody:
Extract the IgG in mouse or pig serum with saturated ammonium sulfate method, get 1 part of serum and add 2 parts of PBS liquid (pH 7.2) and mix, add equal-volume saturated ammonium sulfate liquid and mix, put 2h in 4 DEG C of refrigerators, at 4 DEG C, the centrifugal 15min of 10000r/min, abandon supernatant; With appropriate PBS liquid (pH7.2) dissolution precipitation, adding saturated ammonium sulfate liquid to its ultimate density is 33%, put 2h in 4 DEG C of refrigerators, centrifugal 15min under 4 DEG C, 10000r/min condition, abandons supernatant, with a small amount of PBS liquid (pH7.2) dissolution precipitation, put in 4 DEG C of refrigerators with PBS liquid (pH7.2) dialyzed overnight, change liquid 2～3 times, centrifugal 15min under 4 DEG C, 10000r/min condition, collect supernatant, measure its protein concentration with ultraviolet spectrophotometer.With 50 μ g～100 μ g(IgG)/kg body weight is through subcutaneous or intramuscular injection negative antibody Healthy Sheep or rabbit 3～4 times, last immunity is after 20 days, venous blood collection, measure its serum antibody titer more than 1:2000 with ELISA, heart blood sampling or arteria carotis bloodletting, collect its hyper-immune serum, and its extracting method of IgG(that extracts the anti-mouse of sheep (rabbit) or pig with saturated ammonium sulfate method is identical with said extracted mice serum IgG, no longer repeat), for the preparation of the contrast trace of test strips of the present invention.
2. the preparation of ApxIV monoclonal antibody (Mi):
Every use 50 μ g～100 μ g ApxIV antigen immune Balb/c are mouse three times, every minor tick 15～30d; 3～4d after booster immunization for the third time, by the bloodletting of immune mouse eyeball, draws neck lethal, and with 75% alcohol-pickled 5～10min, aseptic its spleen of getting, shreds and through 100 order nylon net filters, the centrifugal 10min of 1000r/min, collects splenocyte; By 1 × 10
8individual splenocyte and 2～5 × 10
7individual NS0 myeloma cell mixes, 1000r/min is centrifugal, and 10min abandons supernatant, the centrifuge tube that contains sedimentation cell is placed in to the water of 37 DEG C, and slowly add 0.7～1ml, 40%～50% PEG4000(pH 8.5～9.0) effect 1min, then slowly add serum-free 1640 nutrient culture media 15ml, to stop the effect of PEG, 37 DEG C of water-bath 5～10min, 1000r/min is centrifugal, and 10min abandons supernatant, cell precipitation is resuspended in to HAT and selects in nutrient culture media, and add
96 well culture plates (100 μ l/ hole, μ l～200), are placed in 37 DEG C of 5% CO
2in incubator, cultivate.Cultivate after 7～10d, with the coated 96 hole ELISA Plate of pathogen specific antigen of the purifying of 5 μ g～10 μ g/ml, detect the culture supernatant of hybridoma with enzyme linked immunosorbent assay (ELISA), picking strong positive cell clone (OD
450>=0.5), carry out the limiting dilution assay cloning of continuous three times, obtain positive hybridoma cell strain, its chromosome number is 92～98, and the monoclonal antibody of its secretion can specific recognition ApxIV, and not with other toxin generation cross reaction, affinity constant reaches 10
9 ~ 10, light chain subtype is к or λ, heavy chain hypotype is IgG
1, IgG
2a, IgG
2b, IgG
3; The pairing monoclonal antibody obtaining, for making gold mark monoclonal antibody body glass wool or detecting trace.
3. the preparation of gold mark monoclonal antibody glass wool:
Utilize sodium citrate reduction method for preparing nanometer level gold grain: in 0.01～0.05% aqueous solution of chloraurate of 50～100ml boiling, add 0.5～2% citric acid three sodium solution of 2～4ml, obtain the nanometer grade gold particle of diameter 15nm left and right.With the K of 0.1mol/L
2cO
3adjust pH to 8.5～9.5 of gold grain solution, mark with 1:1000～1300 adds in the aurosol of pH8.5～9.5 than by monoclonal antibody to be marked, after mark 10min, add 20% PEG10000 to ultimate density be 0.05%, 4 DEG C, the centrifugal 20min of 1500～3000r/min, remove unconjugated gold grain particle, 4 DEG C, the centrifugal 1h of 15000r/min, abandon supernatant, obtain after golden labeling antibody potpourri, with propylene glucosan S-400 column chromatography, separation and purification gold labeling antibody, the golden labeling antibody of acquisition.By the golden labeling antibody of 1:100～500 dilution, be adsorbed in processed glass cotton 4 DEG C of low-temperature vacuum dryings, preparation gold mark monoclonal antibody glass wool.
4. the preparation of ApxIV polyclonal antibody (Ci):
The preparation of ApxIV polyclonal antibody (Ci).Adopt repeatedly immunity inoculation negative antibody health pig of ApxIV antigen.Last immunity posterior vein blood sampling in 20 days, measure its serum antibody titer more than 1:2000 with ELISA, heart blood sampling or arteria carotis bloodletting, collect its hyper-immune serum, extract IgG antibody (method is identical with the extraction of mice serum IgG, no longer repeats) in serum with saturated ammonium sulfate method.
Gold mark resists the preparation with the how anti-glass wool of gold mark more, identical with the preparation method of gold mark monoclonal antibody glass wool, no longer repeats.Refer to the content 3 in embodiment.
5. test strip of the present invention detects principle
When test strip test lead of the present invention inserts after sample solution to be checked, solution to be checked drives ApxIV to be checked to enter golden labeling antibody fibrage by siphon, and spread to handle end along nitrocellulose filter with together with golden labeling antibody (Mi or Ci) wherein, the final handle end absorbent material layer that infiltrates, in diffusion process, golden labeling antibody can be combined with corresponding ApxIV to be checked, can on cellulose membrane, be detected pairing monoclonal antibody or the how anti-interception of trace in conjunction with the ApxIV of golden labeling antibody, in the time containing tested ApxIV in sample liquid, there is a henna detection line; The anti-mouse of sheep or rabbit or anti-pig IgG can be marked monoclonal antibody or many anti-bindings with corresponding gold, occur 1 brownish red control line.In the time not containing ApxIV in sample liquid to be checked, test strips only demonstrates a brownish red control line; In the time not having control line to show on cellulose membrane, show that test strips lost efficacy.
6. the detection method of operating of test strip of the present invention
(1) detect the processing of sample: get disease pig pathological tissues, 1:1～5 add physiological saline and shred with scissors, and leachate is sample to be checked, and sick pig whole blood or serum are sample to be checked after adding the dilution of physiological saline 1:1～5.
(2) detect operation: test strip sample end of the present invention is inserted in sample liquid to be checked, and insertion depth is no more than mark line 9, after approximately 30 seconds, takes out test strips, horizontal positioned approximately 1～5 minute, simultaneously observations.
(3) result is judged: if only demonstrate a brownish red control line C on test strip cellulose membrane, represent that testing result is negative, illustrate in test sample not containing ApxIV; If there is control line C in the cellulose membrane in test strip, detect trace place and occur a detection line, represent that testing result is positive, in sample to be checked, contain ApxIV; If do not have control line C to show on cellulose membrane, show that test strips lost efficacy.
Embodiment mono-: the test strip of actinobacillus pleuropneumoniae endotoxin (ApxIV)
Referring to Fig. 1 and Fig. 2, in figure, 1 is supporting layer, make by hard plastic strip of foil, 2 is the sample adsorbing fiber layer of test lead, make with glass wool, 3 is golden labeling antibody fibrage, absorption has the glass wool of the monoclonal antibody of the anti-ApxIV of nanometer grade gold particle marker, prepare its gold mark monoclonal antibody glass wool according to the preparation method described in above-mentioned embodiment 3, 4 is cellulose rete, employing nitrocellulose filter is made, 5 is absorbent material layer, make with thieving paper, to number 2, 3, 4, 5 each layers stick on hard plastic strip of foil 1 from left end test lead to the right side, intersection crosses one another overlapping each other.On cellulose nitrate rete 4, the 6 detection trace T for the pairing monoclonal antibody solution printing with anti-ApxIV, 7 is the contrast trace C printing with sheep or rabbit anti-mouse igg solution, detects trace and contrast trace is orthoscopic or oblique line formula, two kinds of trace bands arrange the array configuration forming be "
||", "
//", "
?" in any.8-1 covers test lead sample adsorbing fiber layer 2 and golden labeling antibody fibrage 3 white diaphragm above; on the corresponding diaphragm 8-1 of 2 and 3 intersections position, be partial to sample adsorbing fiber layer 2 one side 0.5cm place and be printed on mark line 9; 9 right-hand member is printed on arrow and max printed words, absorbent material layer 5(handle end) on be coated with other color (as yellow) diaphragm 8-2.
The preparation of testing sample solution and detection operation steps, identical with the detection method of operating in embodiment 6, no longer repeat.
Embodiment bis-: the test strip of actinobacillus pleuropneumoniae endotoxin (ApxIV), basic identical with embodiment mono-, difference is:
The 3 use absorption of gold labeling antibody fibrage have the glass wool of the polyclonal antibody of the anti-ApxIV of gold grain mark to make, and prepare its gold mark polyclonal antibody glass wool according to the preparation method described in above-mentioned embodiment 3; On cellulose nitrate rete 4, the 6 detection trace T for the monoclonal antibody solution printing with anti-ApxIV, 7 is to print contrast trace C with the IgG solution of sheep or the anti-pig of rabbit, and it is " || ", " // ", " \ any in \ " that two kinds of trace bands are arranged the array configuration forming.Other comprises that detection sample preparation, method of operating and result judgement etc. are all identical with the method for operating in embodiment 6, no longer repeats.
Claims (8)
1. one kind is detected the test strip of actinobacillus pleuropneumoniae endotoxin (ApxIV), this test strips contains supporting layer and adsorbed layer, supporting layer is the lamella not absorbing water, adsorbed layer is attached on supporting layer, adsorbed layer is followed successively by the absorbent material layer of sample fiber layer, golden labeling antibody fibrage, cellulose rete and handle end from test lead, on cellulose rete, be prepared with and detect trace and contrast trace, it is characterized in that golden labeling antibody fibrage absorption has the anti-Apx of nanometer grade gold particle marker
monoclonal antibody, detect trace anti-Apx
pairing monoclonal antibody or polyclonal antibody print, the polyclonal antibody of sheep or the anti-mouse IgG of rabbit or staphylococcal protein A (SPA) are printed for contrast trace.
2. test strips according to claim 1, is characterized in that golden labeling antibody fibrage absorption has the anti-Apx of nanometer grade gold particle marker
monoclonal antibody, detects the anti-Apx of trace
pairing monoclonal antibody or polyclonal antibody print, the polyclonal antibody of sheep or the anti-mouse IgG of rabbit or staphylococcal protein A (SPA) are printed for contrast trace, or the absorption of golden labeling antibody fibrage has the anti-Apx of nanometer grade gold particle marker
polyclonal antibody, detect trace anti-Apx
monoclonal antibody preparation, contrast staphylococcal protein A (SPA) or how anti-preparation of anti-pig IgG for trace.
3. test strips according to claim 1 and 2, is characterized in that detecting the anti-Apx of trace
pairing monoclonal antibody preparation use anti-Apx
pairing monoclonal antibody solution preparation; Detect the anti-Apx of trace
polyclonal antibody preparation be and use anti-Apx
polyclonal antibody preparation.
4. test strips according to claim 1, is characterized in that supporting layer the hard plastic slip or the cardboard bar that do not absorb water make; Test lead sample adsorbing fiber layer is made with glass wool; Gold labeling antibody fibrage is made with glass wool and golden labeling antibody, and golden labeling antibody can be monoclonal antibody or polyclonal antibody.
5. test strips according to claim 1, is characterized in that cellulose rete nitrocellulose filter or pure cellulose film or carboxylation cellulose membrane or polyvinylidene fluoride PVDF cellulose membrane make.
6. test strips according to claim 1, is characterized in that absorbent material layer thieving paper makes.
7. test strips according to claim 1, is characterized in that detecting trace and contrast trace is orthoscopic or oblique line formula, on cellulose rete, contain three and detect traces and one contrast trace, the spread pattern that detects trace and contrast trace be "
||", "
//", "
?" in any.
8. test strips according to claim 1; it is characterized in that containing layer protective layer above test strips adsorbed layer; protective seam is attached on adsorbed layer; on test lead sample adsorbing fiber layer, golden labeling antibody fibrage and absorbent material layer, be coated with diaphragm; on the test lead sample adsorbing fiber layer diaphragm corresponding with golden labeling antibody fibrage intersection, be printed with sample mark line, this mark line deflection test lead sample adsorbing fiber layer about 0.5cm of one side place.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310083779.3A CN103941013B (en) | 2013-03-15 | 2013-03-15 | Actinobacillus pleuropneumoniae endotoxin Rapid detection test strip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310083779.3A CN103941013B (en) | 2013-03-15 | 2013-03-15 | Actinobacillus pleuropneumoniae endotoxin Rapid detection test strip |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103941013A true CN103941013A (en) | 2014-07-23 |
| CN103941013B CN103941013B (en) | 2016-03-16 |
Family
ID=51188766
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310083779.3A Expired - Fee Related CN103941013B (en) | 2013-03-15 | 2013-03-15 | Actinobacillus pleuropneumoniae endotoxin Rapid detection test strip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103941013B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105116142A (en) * | 2015-08-05 | 2015-12-02 | 上海拜攸生物科技有限公司 | Novel bacterial endotoxin detection test paper and detection method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004052925A2 (en) * | 2002-12-09 | 2004-06-24 | Imperial College Innovations Limited | Actinobacillus pleuropneumoniae virulence genes |
| CN101265457A (en) * | 2007-07-20 | 2008-09-17 | 华中农业大学 | Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof |
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
| CN101949934A (en) * | 2010-09-04 | 2011-01-19 | 扬州大学 | Early infection detection kit of monoclonal antibody-mediated pig pleuropneumoniae |
-
2013
- 2013-03-15 CN CN201310083779.3A patent/CN103941013B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004052925A2 (en) * | 2002-12-09 | 2004-06-24 | Imperial College Innovations Limited | Actinobacillus pleuropneumoniae virulence genes |
| CN101265457A (en) * | 2007-07-20 | 2008-09-17 | 华中农业大学 | Vaccine for differentiating porcine actinobacillus pleuropneumonia serum 7-type double-gene deletion mutant of vaccine immunity and virus infection animal and application thereof |
| CN101339192A (en) * | 2008-08-28 | 2009-01-07 | 河南省农业科学院 | Test paper for one-step detection for pig virus diarrhoea disease pathogen |
| CN101949934A (en) * | 2010-09-04 | 2011-01-19 | 扬州大学 | Early infection detection kit of monoclonal antibody-mediated pig pleuropneumoniae |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105116142A (en) * | 2015-08-05 | 2015-12-02 | 上海拜攸生物科技有限公司 | Novel bacterial endotoxin detection test paper and detection method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103941013B (en) | 2016-03-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101339192A (en) | Test paper for one-step detection for pig virus diarrhoea disease pathogen | |
| CN104407137A (en) | Test paper for identifying and detecting virulent strain and low virulent strain of hog cholera virus | |
| CN205506834U (en) | Short -term test aflatoxin B1 and zearalenone's colloidal gold test paper | |
| CN104459144A (en) | Porcine pseudorabies virus virulent strain and vaccine strain identification and detection test paper | |
| CN105137073A (en) | Bovine Brucella colloidal gold antibody detection test paper strip | |
| CN103266088A (en) | H7 subtype avian influenza virus monoclonal antibody of and kit | |
| CN106018799A (en) | Test paper for salmonella and preparation method of test paper | |
| CN104459142A (en) | Test paper for distinguishing virulent strain from attenuated strain of newcastle disease virus | |
| CN104374914B (en) | A kind of pseudomonas putida test strip and preparation method thereof | |
| CN102994455A (en) | Monoclonal antibody and kit for cucumber green mottle mosaic viruses (CGMMVs) | |
| CN101424689B (en) | Bluetongue virus detection test paper | |
| CN103941013B (en) | Actinobacillus pleuropneumoniae endotoxin Rapid detection test strip | |
| CN101339193B (en) | Eperythrozoonosis rapid diagnosis test paper | |
| CN103941001B (en) | Haemophilus parasuis infection Rapid detection test strip | |
| CN104101707A (en) | 2, 4-dichlorphenoxyacetic acid residue rapid detection test paper strip | |
| CN101074955B (en) | Immune chromatography test paper for inspecting legionella pneumophilia antibody and its production | |
| CN103941014B (en) | Swine enzootic pneumonia, porcine contagious pleuropneumonia and swine plague three test strip | |
| CN204495834U (en) | Detect the test strips of chicken trachitis virus | |
| CN204495838U (en) | A kind of test strips detecting Avianreovirus | |
| CN103940991B (en) | Pig salmonella heat-stable toxin Rapid detection test strip | |
| CN103235129B (en) | Marek's disease virus and J subgroup avian leucosis virus fast joint inspection test strips | |
| CN103941003B (en) | Pig Listeria monocytogenes, pig necrobacillus and pig Bacillus tularensis three test strip | |
| CN204287191U (en) | The two-in-one immune colloid gold Rapid detection test strip of a kind of rice leaf spot bacteria and xanthomonas oryzae pv. oryzicola | |
| CN104777302A (en) | Immunochromatographic assay test strip for bacterial fruit blotch of melons | |
| CN103217527B (en) | Marek's disease virus and avian reticuloendotheliosis virus fast joint inspection test strips |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160316 |