CN110498842A - Peste des petits ruminants diagnostic kit - Google Patents
Peste des petits ruminants diagnostic kit Download PDFInfo
- Publication number
- CN110498842A CN110498842A CN201810480175.5A CN201810480175A CN110498842A CN 110498842 A CN110498842 A CN 110498842A CN 201810480175 A CN201810480175 A CN 201810480175A CN 110498842 A CN110498842 A CN 110498842A
- Authority
- CN
- China
- Prior art keywords
- polypeptide
- seq
- sequence shown
- des petits
- peste des
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- 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
- G01N33/56983—Viruses
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
- C12N2760/00011—Details
- C12N2760/18011—Paramyxoviridae
- C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
- C12N2760/18422—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- 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
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/115—Paramyxoviridae, e.g. parainfluenza virus
- G01N2333/12—Mumps virus; Measles virus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2469/00—Immunoassays for the detection of microorganisms
- G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The present invention relates to peste des petits ruminants diagnostic kits.Detection kit of the invention includes substrate, and particular polypeptide or the particular polypeptide combination being independently connected in the substrate.
Description
Technical field
The invention mainly relates to diagnostic kit for animals and diagnostic methods.Specifically, the present invention relates to detections pair
As biological source biological sample in the presence or absence of anti-PPR virus antibody (IgG) kit.
Background technique
Peste des petits ruminants (peste des petits ruminants, PPR) be caused by PPR virus it is small anti-
A kind of Acute exposure communicable disease of hay animal, belongs to a kind of animal epidemic, and the clinical manifestation of the disease is similar to rinderpest, therefore
Referred to as pseudo- rinderpest, it is small ruminate beast vacation rinderpest, be also called and small ruminate beast pest, sheep pest, contagious ecthyma shape gastritis, pneumonia enteritis
Syndrome etc..PPR virus category paramyxovirus section Morbillivirus.Virus is in pleomorphism, usually coarse spherical shape.
PPR virus is the virus for having cyst membrane, and resistance is lower under natural environment, and 50 DEG C can inactivate for 60 minutes；
It is inactivated under the conditions of pH<4.0 or pH>11.0；But it can survive the long period in refrigeration and freezing tissue.Alcohol, ether and common
Detergent can kill virus, and phenol and 2% NaOH are effective disinfectants.
The disease is named as " small to ruminate epidemic disease " after the African western Ivory Coast is broken out for the first time in nineteen forty-two, is now popular in
Interior jar (unit of capacitance), Ghana and the Arabia Peninsula in Africa and middle east, once had case report in Asia, India, Bangladesh and Israel
It accuses, it is to occur on July 25th, 2007 to send out in the village Long Menka, the Ritu County township Re Bang, Tibet Autonomous Region that, which there is the small epidemic disease of ruminating of record in China,
Raw goat peste des petits ruminants epidemic situation, dead 262.The disease is mainly seen in the small ruminants such as sheep, goat, antelope, but goat
Than more serious when morbidity.Ox, pig can infect, but usually subclinical process.Infected animal can produce viremia virusemia, virus
It is distributed widely in various tissues, and can be excreted with various secretion and excreta, becomes the infection sources.Virus is by directly connecing
Touching, is entered animal body by approach such as nose, mouths and is propagated.The disease can pass through air borne between the animal of close contact.This disease
Whole year can occur, but usually based on rainy season and dry and cold season.In morbidity epidemic place, morbidity and mortality be increased significantly.Often
Occurred in the form of scattered epidemic place, after certain times are in outbreak of epidemic, then has the mitigation phase of a 5-6.But old epidemic-stricken area
Epidemic place is then constant to be occurred and is increasing.Clinical symptoms summarize are as follows: high continued fever, conjunctivitis, necrotic stomatitis, pneumonia,
Diarrhea.Dissect symptom are as follows: the naked eyes lesion such as visible conjunctivitis, necrotic stomatitis, several cases can spread to hard palate and bottleneck throat.
Seldom there is lesion in cud, reticulum, omasum, and rotten to the corn lesion then often occurs in abomasum, and the surface of a wound is in blood red.Enteron aisle has rotten to the corn or goes out
Blood variation, especially can usually find characteristic streak-like hemorrhage or zebra batten line in colon and rectum junction.The enlargement of lymph node
Greatly, spleen has gangrenosum acne lesion.There is blood spots in concha, larynx, tracheae etc..
Currently, effective ways there is no to treat peste des petits ruminants, vaccine inoculation, epidemic situation can only be taken to slaughter and periodically after occurring
The method of sero monitoring is controlled.Therefore, the serodiagnosis of the disease and the assessment of immune effect of vaccine seem with monitoring
It is particularly important.
World Organization for Animal Health recommends the PPR virus antibody detection method used mainly to have virus to neutralize examination
Test (VNT) and enzyme-linked immunosorbent assay (ELISA).Wherein, the testing result of virus neutralization tests method is accurate, and being that detection is small ruminates
The goldstandard of epizootic disease virus, but this method detection time is long and is unsuitable for detecting great amount of samples.In contrast, enzyme-linked immunosorbent assay
Specificity and sensibility is higher, detection time is shorter than virus neutralization tests and therefore the suitable a large amount of samples of detection are answered extensively
Detection for PPR virus antibody.
Enzyme-linked immunosorbent assay method for PPR virus antibody test mainly includes competitive enzyme-linked immune measurement
(c-ELISA), enzyme-linked immunosorbent assay (b-ELISA) and indirect enzyme-linked immunosorbent measurement (indirect ELISA) are blocked.C-ELISA and b-
The specificity and sensibility of ELISA is relatively high, is by clinical generally accepted PPR virus antibody detection method, example
C-ELISA method is used if the peste des petits ruminants diagnostic kit in the French laboratory BIRAD general in the world.But
C-ELISA and b-ELISA is both needed to using monoclonal antibody, this causes testing cost to greatly improve；Moreover, c-ELISA and b-
The cumbersome of ELISA, detection time long (although having been shortened relative to VNT), criterion are complicated.On the other hand, traditional
Indirect ELISA use from PPR virus intact proteins (such as H protein, N protein, F protein etc.) or recombination egg
White as envelope antigen to detect the antibody in serum, cost is lower than c-ELISA and b-ELISA；But since this method makes
It uses intact proteins as antigen, is easy to happen the wrong identification and non-specific identification of antibody, therefore, specificity and sensibility
All not as good as c-ELISA and b-ELISA.
Therefore, it is necessary to develop, testing cost is low, detection time is short, easy to operate, specific and high sensibility small anti-
Hay epizootic disease diagnostic kit.
Summary of the invention
In view of above-mentioned problems of the prior art, low, detection that the purpose of the present invention is to provide a kind of testing costs
Time short, easy to operate, specific and high sensibility peste des petits ruminants diagnostic kit, and can be used in preparing the reagent
The polypeptide or polypeptides in combination of box.
Indirect ELISA method based on antigen epitope polypeptide uses the antigen epitope polypeptide (single of 20 amino acid or so
Polypeptide usually only includes an epitope) it is used as envelope antigen.Inventor's discovery: the sensibility of this method is often relatively low, single
The sensibility of polypeptide does not exceed 50% generally；And the polypeptide that sensitivity is high, false positive rate are also high, i.e., it is specific low.If
The specificity and sensibility of detection can be improved simultaneously by certain mode, so that it may the shortcomings that overcoming traditional indirect ELISA,
Developing specificity and sensibility can match in excellence or beauty c-ELISA and b-ELISA, even higher peste des petits ruminants diagnostic kit.
Inventor has made intensive studies to solve above-mentioned technical problem, as a result, it has been found that following polypeptides in combination 1.When with this
It at least any one in following polypeptides in combination 1 when having response as index to the biological sample in object organisms source, can be with
96.0% sensibility and 87.1% specific diagnosis peste des petits ruminants, completely can be with c-ELISA method and the side b-ELISA
Method matches in excellence or beauty.
Therefore, the present invention includes:
1. a kind of peste des petits ruminants diagnostic kit comprising substrate, and under being individually secured in the substrate
State polypeptides in combination 1；
Polypeptides in combination 1
Sequence shown in SEQ ID NO:1 is the polypeptide of N50:SAEALFRLQAMAKILEDQEE,
Sequence shown in SEQ ID NO:2 is the polypeptide of N49:SSQNPREAQRSAEALFRLQA,
Sequence shown in SEQ ID NO:3 is the polypeptide of F54:LKPDLTGTSKSYVRSL,
Sequence shown in SEQ ID NO:4 is the polypeptide of F13:VATAAQITAGVALHQSLMNS,
Sequence shown in SEQ ID NO:5 is the polypeptide of N41:TGDERTVRGTGPRQAQVSFL,
Sequence shown in SEQ ID NO:6 is the polypeptide of F10:TKNVRPIQTLTPGRRTRRFV, and
Sequence shown in SEQ ID NO:7 is the polypeptide of N9:VESPGQIQRITDDPDVSIR.
2. being used for the biology by test object biological source according to peste des petits ruminants diagnostic kit described in item 1
Whether the antibody (IgG) that whether there is anti-PPR virus in sample diagnoses object organisms by PPR virus sense
Dye or peste des petits ruminants vaccine immunity.
3. according to peste des petits ruminants diagnostic kit described in item 2, wherein the object organisms small ruminant.
4. the peste des petits ruminants diagnostic kit according to item 2 or 3, wherein the object organisms are goat or sheep.
5. the peste des petits ruminants diagnostic kit according to any one of item 2~4, wherein the biological sample is complete
Blood, blood plasma or serum.
6. purposes of following polypeptides in combination 1 in preparation peste des petits ruminants diagnostic kit；
Polypeptides in combination 1
Sequence shown in SEQ ID NO:1 is the polypeptide of N50:SAEALFRLQAMAKILEDQEE,
Sequence shown in SEQ ID NO:2 is the polypeptide of N49:SSQNPREAQRSAEALFRLQA,
Sequence shown in SEQ ID NO:3 is the polypeptide of F54:LKPDLTGTSKSYVRSL,
Sequence shown in SEQ ID NO:4 is the polypeptide of F13:VATAAQITAGVALHQSLMNS,
Sequence shown in SEQ ID NO:5 is the polypeptide of N41:TGDERTVRGTGPRQAQVSFL,
Sequence shown in SEQ ID NO:6 is the polypeptide of F10:TKNVRPIQTLTPGRRTRRFV, and
Sequence shown in SEQ ID NO:7 is the polypeptide of N9:VESPGQIQRITDDPDVSIR.
7. according to purposes described in item 6, wherein the peste des petits ruminants diagnostic kit is used for through test object biology
It whether there is the antibody (IgG) of anti-PPR virus in the biological sample in source, whether diagnosis object organisms are ruminated by small
Epizootic disease virus infection or peste des petits ruminants vaccine immunity.
8. according to purposes described in item 7, wherein the object organisms small ruminant.
9. the purposes according to item 7 or 8, wherein the object organisms are goat or sheep.
10. the purposes according to any one of item 7~9, wherein the biological sample is whole blood, blood plasma or serum.
Aforementioned polypeptides and kit, which can be used for whether there is in the biological sample of test object biological source, resists small ruminate
The antibody (IgG) of epizootic disease virus.In general, the PPR virus antibody (IgG) in biological sample is due to object organisms quilt
PPR virus is infected or is generated by peste des petits ruminants vaccine immunity, and therefore, aforementioned polypeptides and kit can be used
In diagnosis object organisms whether by PPR virus infection or peste des petits ruminants vaccine immunity.
The specific embodiment of invention
Firstly, the present invention provides following polypeptides in combination 1:
Polypeptides in combination 1
Sequence shown in SEQ ID NO:1 is the polypeptide of N50:SAEALFRLQAMAKILEDQEE,
Sequence shown in SEQ ID NO:2 is the polypeptide of N49:SSQNPREAQRSAEALFRLQA,
Sequence shown in SEQ ID NO:3 is the polypeptide of F54:LKPDLTGTSKSYVRSL,
Sequence shown in SEQ ID NO:4 is the polypeptide of F13:VATAAQITAGVALHQSLMNS,
Sequence shown in SEQ ID NO:5 is the polypeptide of N41:TGDERTVRGTGPRQAQVSFL,
Sequence shown in SEQ ID NO:6 is the polypeptide of F10:TKNVRPIQTLTPGRRTRRFV, and
Sequence shown in SEQ ID NO:7 is the polypeptide of N9:VESPGQIQRITDDPDVSIR.
In the indirect ELISA method based on antigen epitope polypeptide, the sensibility for being usually no more than 50% of single polypeptide
It compares, is having response as index the biological sample in object organisms source using at least any one in the polypeptides in combination 1
In the case of, it can be with 96.0% sensibility and 87.1% specific diagnosis peste des petits ruminants.
In this specification, sensibility refers to: in the positive sample with the confirmation of " goldstandard " method, being measured as by other methods
The ratio of positive sample.Specificity refers to: in the negative sample with the confirmation of " goldstandard " method, being measured as feminine gender by other methods
The ratio of sample.For the detection of PPR virus antibody, " goldstandard " of the art is that virus neutralizes examination
Test (VNT).
Aforementioned polypeptides combination can be used as detection probe be used to prepare in the biological sample of test object biological source whether
There are the kits of the antibody of anti-PPR virus (IgG).In general, the PPR virus antibody in biological sample
It (IgG) is to be infected due to object organisms by PPR virus or generated by peste des petits ruminants vaccine immunity, on
It states polypeptides in combination and whether kit can be used for diagnosing object organisms by PPR virus infection or peste des petits ruminants epidemic disease
Seedling is immune.
Therefore, the present invention also provides a kind of peste des petits ruminants diagnostic kits comprising substrate, and be independently connected to
Aforementioned polypeptides combination 1 in the substrate.
In the present specification, substrate is normally solid, can be one, be also possible to it is multiple, but preferably one, i.e., entirely
Portion's polypeptide is independently connected in same substrate.In the present invention, substrate is not particularly limited, as long as solid or insoluble
Material support.The connection of polypeptide and substrate can be using well known to a person skilled in the art the connections of polypeptide and solid material
Method carries out.
In the present specification, the object organisms are preferably small ruminant, more preferably goat or sheep.
In the present specification, the biological sample can be whole blood, blood plasma or serum.
In the case where stating kit diagnosis peste des petits ruminants in use, as any in the polypeptides in combination 1 or one
When the above polypeptide has response to the biological sample in object organisms source, determine that the object organisms are infected by PPR virus
Or peste des petits ruminants vaccine immunity (i.e. positive)；Conversely, determining the object organisms not by PPR virus infection or small anti-
Hay epizootic disease vaccine immunity (i.e. negative).
In the present specification, " response " refers to: signal-to-noise ratio (SNR) is greater than or equal to 2, wherein signal-to-noise ratio=(polypeptide point letter
Number value-negative control point signal value)/negative control point signal value.
Embodiment
1. the preparation and confirmation of polypeptide
The polypeptide of NO:1~7 SEQ ID is synthesized by gill biochemistry (Shanghai) Co., Ltd., and is carried out by mass spectrum to it
Confirmation.Wherein,
The sequence of SEQ ID NO:1 is N50:SAEALFRLQAMAKILEDQEE；
The sequence of SEQ ID NO:2 is N49:SSQNPREAQRSAEALFRLQA；
The sequence of SEQ ID NO:3 is F54:LKPDLTGTSKSYVRSL；
The sequence of SEQ ID NO:4 is F13:VATAAQITAGVALHQSLMNS；
The sequence of SEQ ID NO:5 is N41:TGDERTVRGTGPRQAQVSFL；
The sequence of SEQ ID NO:6 is F10:TKNVRPIQTLTPGRRTRRFV；
The sequence of SEQ ID NO:7 is N9:VESPGQIQRITDDPDVSIR.
2. the preparation of kit (polypeptide chip) 1
Kit 1
Polypeptide chip is the solution difference point sample by the polypeptide of NO:1~7 SEQ ID in solid support material --- " 0+X "
Be prepared on film (iPDMS film) (while one goat IgG of point sample is used as feminine gender as positive quality control point and a PB point
Quality Control point)." 0+X " film is that the initiator with olefin-terminal, surface initiated polymerization is added to dimethyl silicone polymer
In material, then in the three-dimensional structure in a manner of heat cross-linking (si-h bond bonding) fixed to dimethyl silicone polymer, obtain
A kind of material.Its manufacturing process can be found in published International patent WO2014/044184.
3. being detected with kit
Polypeptide chip is placed in room temperature 20min, visually observes chip surface, there will be the polypeptide chip of defect to eliminate, simultaneously
The correct direction for determining polypeptide chip, is sandwiched in tongue, and the number of chip is carried out according to the quantity of serum, then in qualified polypeptide
TBST (0.4M Tris-HCl, 2.74M NaCl, 2%Tween20, pH7.2 ± 0.2) is filled in chip, is placed at room temperature for 2-
4min checks for that in the dead of night, the polypeptide chip of leakage being discarded.The polypeptide chip of vacancy is filled up, operation is same as above.It is qualified to examine
Polypeptide chip be sandwiched on tongue, every 8 chips are one group, and with TBST rinse 2 times, watering can elution a, hole Kong Jinyi goes out, and are formed
Flowing, is finally gently patted on gauze, another one takes lid to dry, but it is noted that holding polypeptide chip surface wettability, standby
With.
Serum is diluted 50 times (+245 μ l serum dilutions of 5 μ l serum sample) in super-clean bench to number respectively, is shaken
It mixes, chip number is made to number consistent with each other, the loading on the chip of rinse with serum, 200 holes μ l/ avoid generating gas
Bubble, is added the 200 μ l of serum after diluting, room temperature, and shaking table 150rpm is incubated for 30min.
Liquid is abandoned, 4 times (method is same as above) is eluted, drying is added secondary antibody (rabbit-anti goat IgG), room temperature, and shaking table 150rpm is incubated
Educate 30min.
Liquid is abandoned, lid is abandoned, is eluted 4 times, drying, every 8 chips are one group, add luminescent solution (Thermo, the Prod# in the hole 20ul/
37074) 2min, is exposed.Data are acquired using GenePix Pro6.0.
For every a serum, whether each polypeptide counted in the kit respectively has response (that is, signal-to-noise ratio
(SNR) it is more than or equal to 2), and determined.
For mentioned reagent box 1, when detect any one or one in polypeptide shown in NO:1~7 SEQ ID with
On when having response, be determined as the peste des petits ruminants positive；Conversely, being determined as feminine gender.
Wherein, signal-to-noise ratio=(polypeptide point signal value-negative control point signal value)/negative control point signal value.Polypeptide point
Signal value refers to the chemiluminescence intensity value for the polypeptide point that software is read, and negative control point signal value refers to the feminine gender that software is read
The chemiluminescence intensity value of control point.
For 755 parts of goats and sheep serum sample from China regions, it is respectively adopted and is prepared in above-mentioned steps 2
Kit (the step of pressing above-mentioned 3) and virus neutralization tests (VNT, by OIE (2010) " PPR virus neutralization test "
The method) it is detected, testing result is as follows.
Table 1
Sensibility=383/399*100%=96.0%
Specificity=310/356=87.1%
As known from the above, it (and is tested using VNT method that the sensibility of kit 1, which is 96.0%, specificity is 87.1%
Card is verified using contrast agents cassette method), it is sufficient to the kit to match in excellence or beauty using c-ELISA method or b-ELISA method.
Sequence table
<110>China Veterinery Drug Inspection Office
Luoyang Zhong Ke Bioarray Solutions Ltd.
<120>peste des petits ruminants diagnostic kit
<130> PB00107
<141> 2018-04-24
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 1
Ser Ala Glu Ala Leu Phe Arg Leu Gln Ala Met Ala Lys Ile Leu Glu
1 5 10 15
Asp Gln Glu Glu
20
<210> 2
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 2
Ser Ser Gln Asn Pro Arg Glu Ala Gln Arg Ser Ala Glu Ala Leu Phe
1 5 10 15
Arg Leu Gln Ala
20
<210> 3
<211> 16
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 3
Leu Lys Pro Asp Leu Thr Gly Thr Ser Lys Ser Tyr Val Arg Ser Leu
1 5 10 15
<210> 4
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 4
Val Ala Thr Ala Ala Gln Ile Thr Ala Gly Val Ala Leu His Gln Ser
1 5 10 15
Leu Met Asn Ser
20
<210> 5
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 5
Thr Gly Asp Glu Arg Thr Val Arg Gly Thr Gly Pro Arg Gln Ala Gln
1 5 10 15
Val Ser Phe Leu
20
<210> 6
<211> 20
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 6
Thr Lys Asn Val Arg Pro Ile Gln Thr Leu Thr Pro Gly Arg Arg Thr
1 5 10 15
Arg Arg Phe Val
20
<210> 7
<211> 19
<212> PRT
<213>artificial sequence (Artificial Sequence)
<400> 7
Val Glu Ser Pro Gly Gln Ile Gln Arg Ile Thr Asp Asp Pro Asp Val
1 5 10 15
Ser Ile Arg
Claims (10)
1. a kind of peste des petits ruminants diagnostic kit is used to whether there is in the biological sample by test object biological source
The IgG antibody of anti-PPR virus, whether diagnosis object organisms are by PPR virus infection or peste des petits ruminants epidemic disease
Seedling is immune.
2. peste des petits ruminants diagnostic kit according to claim 1 comprising substrate, and be individually secured to described
Following polypeptides in combination 1 in substrate；
Polypeptides in combination 1
Sequence shown in SEQ ID NO:1 is the polypeptide of N50:SAEALFRLQAMAKILEDQEE,
Sequence shown in SEQ ID NO:2 is the polypeptide of N49:SSQNPREAQRSAEALFRLQA,
Sequence shown in SEQ ID NO:3 is the polypeptide of F54:LKPDLTGTSKSYVRSL,
Sequence shown in SEQ ID NO:4 is the polypeptide of F13:VATAAQITAGVALHQSLMNS,
Sequence shown in SEQ ID NO:5 is the polypeptide of N41:TGDERTVRGTGPRQAQVSFL,
Sequence shown in SEQ ID NO:6 is the polypeptide of F10:TKNVRPIQTLTPGRRTRRFV, and
Sequence shown in SEQ ID NO:7 is the polypeptide of N9:VESPGQIQRITDDPDVSIR.
3. peste des petits ruminants diagnostic kit according to claim 2, wherein the object organisms small ruminant.
4. peste des petits ruminants diagnostic kit according to claim 2, wherein the object organisms are goat or sheep.
5. peste des petits ruminants diagnostic kit according to claim 2, wherein the biological sample be whole blood, blood plasma or
Serum.
6. purposes of following polypeptides in combination 1 in preparation peste des petits ruminants diagnostic kit；
Polypeptides in combination 1
Sequence shown in SEQ ID NO:1 is the polypeptide of N50:SAEALFRLQAMAKILEDQEE,
Sequence shown in SEQ ID NO:2 is the polypeptide of N49:SSQNPREAQRSAEALFRLQA,
Sequence shown in SEQ ID NO:3 is the polypeptide of F54:LKPDLTGTSKSYVRSL,
Sequence shown in SEQ ID NO:4 is the polypeptide of F13:VATAAQITAGVALHQSLMNS,
Sequence shown in SEQ ID NO:5 is the polypeptide of N41:TGDERTVRGTGPRQAQVSFL,
Sequence shown in SEQ ID NO:6 is the polypeptide of F10:TKNVRPIQTLTPGRRTRRFV, and
Sequence shown in SEQ ID NO:7 is the polypeptide of N9:VESPGQIQRITDDPDVSIR.
7. purposes according to claim 6, wherein the peste des petits ruminants diagnostic kit is used for raw by test object
It whether there is the IgG antibody of anti-PPR virus in the biological sample in object source, whether diagnosis object organisms are ruminated by small
Epizootic disease virus infection or peste des petits ruminants vaccine immunity.
8. purposes according to claim 7, wherein the object organisms small ruminant.
9. purposes according to claim 7, wherein the object organisms are goat or sheep.
10. purposes according to claim 7, wherein the biological sample is whole blood, blood plasma or serum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810480175.5A CN110498842B (en) | 2018-05-18 | 2018-05-18 | Peste des petits ruminants diagnostic kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810480175.5A CN110498842B (en) | 2018-05-18 | 2018-05-18 | Peste des petits ruminants diagnostic kit |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110498842A true CN110498842A (en) | 2019-11-26 |
| CN110498842B CN110498842B (en) | 2021-01-26 |
Family
ID=68585136
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810480175.5A Active CN110498842B (en) | 2018-05-18 | 2018-05-18 | Peste des petits ruminants diagnostic kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110498842B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014044184A1 (en) * | 2012-09-21 | 2014-03-27 | 苏州偲聚生物材料有限公司 | Protein for detecting type 1 diabetes mellitus and partial peptide thereof |
| CN107098979A (en) * | 2017-06-22 | 2017-08-29 | 中国动物疫病预防控制中心 | PPR virus H F fusion proteins and its relevant biological material and application |
| CN107216372A (en) * | 2017-06-27 | 2017-09-29 | 中国农业科学院兰州兽医研究所 | A kind of PPR virus HN Protein Epitopes peptide H362 and its determination, preparation method and application |
-
2018
- 2018-05-18 CN CN201810480175.5A patent/CN110498842B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2014044184A1 (en) * | 2012-09-21 | 2014-03-27 | 苏州偲聚生物材料有限公司 | Protein for detecting type 1 diabetes mellitus and partial peptide thereof |
| CN107098979A (en) * | 2017-06-22 | 2017-08-29 | 中国动物疫病预防控制中心 | PPR virus H F fusion proteins and its relevant biological material and application |
| CN107216372A (en) * | 2017-06-27 | 2017-09-29 | 中国农业科学院兰州兽医研究所 | A kind of PPR virus HN Protein Epitopes peptide H362 and its determination, preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| E. COUACY-HYMANN 等: "Interference in the vaccination of cattle against rinderpest virus by antibodies against peste des petits ruminants (PPR) virus", 《VACCINE》 * |
| 石晓妮 等: "小反刍兽疫病毒H和F蛋白多抗原表位的表达及其反应原性", 《中国兽医科学》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110498842B (en) | 2021-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106018800A (en) | Detecting device for Brucella infection | |
| CN101379398A (en) | Diagnostic formulation for tsutsugamushi disease | |
| Ozan et al. | Serological determination of pestivirus, bluetongue virus and peste des petits ruminants virus in small ruminants in Samsun province of Turkey. | |
| Ren et al. | A brief review of diagnosis of small ruminants brucellosis | |
| CN115873079A (en) | Canine infectious hepatitis virus hexon protein antigen, truncation and application thereof | |
| CN110498842A (en) | Peste des petits ruminants diagnostic kit | |
| Dhliwayo et al. | Seroprevalence of leptospirosis in dogs in urban Harare and selected rural communities in Zimbabwe | |
| CN110221060A (en) | A kind of antigen and its application for early diagnosing haemonchus contortus infection | |
| CN110498845A (en) | Peste des petits ruminants diagnostic kit | |
| Baloch et al. | Study on the Prevalence of Peste Des Petits Virus Antibodies in Caprine and Ovine Through the Contrast of Serological Assessments in Sindh, Pakistan | |
| CN110498844A (en) | Peste des petits ruminants diagnostic kit | |
| CN112213493B (en) | Peste des petits ruminants detection kit capable of being used for distinguishing vaccine immunity from natural infection | |
| CN111537713B (en) | Peste des petits ruminants virus antibody detection kit | |
| Genc et al. | Development of qualitative and quantitative ELISA models for bovine brucellosis diagnosis | |
| CN110498843B (en) | Peste des petits ruminants diagnostic kit | |
| CN110501505A (en) | Peste des petits ruminants diagnostic kit | |
| Aziz et al. | Thin layer immunoassay; an economical approach to diagnose Helicobacter pylori Infection in gastroduodenal ulcer disease patients of Pakistan; a comparative analysis | |
| LU101824B1 (en) | Novel coronavirus testing | |
| CN106198973A (en) | What a kind of simplicity quickly detected B rucella melitensis antibody directly blocks ELISA method | |
| KR102021538B1 (en) | Peptides for diagnosis of mosquito-borne viruses and uses thereof | |
| Lewin-Van Den Broek et al. | Validation and value of an enzyme-linked immunosorbent assay for Helicobacter pylori in primary care | |
| Feliciano et al. | Helicobacter pylori antigen recognition patterns among cuban patients using Helicoblot 2.1 diagnostic system | |
| Duman et al. | Virological and serological investigations of bluetongue virus (BTV) infection in sheep in Konya region | |
| Gospodinova et al. | Comparison of a rapid enzyme-linked immunosorbant assay test with an indirect immunofluorescent antibody test in diagnosing Ehrlichia and Anaplasma infections in dogs | |
| CN115160412A (en) | Peste des petits ruminants virus antibody detection kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |