CN103773846B - In a kind of food and feeds, goose derived components detects lateral flow ELISA test strip kit and application thereof - Google Patents
In a kind of food and feeds, goose derived components detects lateral flow ELISA test strip kit and application thereof Download PDFInfo
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6804—Nucleic acid analysis using immunogens
Abstract
The invention discloses food and feeds become in goose (<i>Geese</i>) preparation method and the application process thereof of nucleic acid rapid detection kit of composition, belong to molecular biology and field of immunology. The present invention combines the high sensitivity of the PCR in detection of nucleic acids, high specific method with the immune colloid gold Fast Detection Technique in immunology detection, by designing unique primer, and primer is carried out to mark, to the target DNA extracting carry out specific amplification and amplified production in developping solution in test strips fixing golden labelled antibody be combined and forms stablize, visible detection band and quality control band, thereby realization detects fast and accurately to goose derived components in food and feeds.
Description
Technical field
The invention belongs to molecular biology and field of immunology, the PCR nucleic acid amplification of goose derived component in food and feeds,And the methods for making and using same of lateral flow immunity colloidal gold test paper strip kit.
Background technology
Moving through being added on for preventing BSE (bovinespongiformencephalopathy, BSE)Beef, ox bone in thing feed are propagated, and European Union in 1994 and the U.S. in 1997 formulated and forbid at 1994 and 1997 respectivelyIn the feed of ruminant, add the regulation of ruminant endogenous binding protein, within 2000, European Union expands this ban to forbid to use toIn the finished mammal of edible detoxification, birds and fish protein composition. The Ministry of Agriculture of China promulgated in 2004In " feed products from animal sources safety and sanitation management method ", provide against and in ruminant feed, use animal derived feedProduct (except milk and milk products), but in animal feed, people mixes ruminant (being mainly ox, sheep) meat and bone warePhenomenon happens occasionally.
In recent years, both at home and abroad on food products market, the phenomenon of mixing the spurious with the genuine such as false mutton, false beef emerges in an endless stream, animal derived foodTaking place frequently of the various food safety affairs such as product doping is adulterated more and more allows the frightened courage of common people's heart quiver. At the beginning of 2013, " the horseflesh in EuropeDisturbance " grow in intensity, make consumer's " Wen Mase change ". American-European afterwards " flesh of fish disturbance " sound again again food securityAlarm bell. Ensure the safety of food and feeds, needs are sensitive, convenient, detection method provides basis accurately.
In traditional food and feeds, animality Components identification is mainly by microscope inspection, immunity based on protein detectionMethod and all kinds of PCR methods based on detection of nucleic acids, also developed near infrared detection method (NIRS) in recent years. Food and feedsThe various animal sources compositions that mix in composition are easy to be damaged in high-temperature process and process, the egg of different genera animalBetween white matter sequence, difference is little, is difficult to difference, and " SDS-PAGE method is differentiated animal derived one-tenth in meat and meat products to patent of inventionPart " (publication number: CN102213720A) provide a kind of SDS-PAGE electrophoretic with protein qualification pig, ox, sheep,The method of the animal protein of goose, fish, but the protein electrophoresis bands of a spectrum of separate sources and processing mode are different, the barrier that brings result to differentiateHinder, be more difficult to carry out for the animal component in feed.
Because the degeneracy feature of gene coded sequence and the existence of non-coding sequence, nucleic acid becomes compared with albumen and more easily detectsThe target molecule of difference, particularly mitochondrial DNA, copy number is high, and existence interspecific difference is main to the analysis of its differential fragmentAuthentication method, these class methods are because of highly sensitive, specificity is good, rapidly, main method comprises common/glimmering for less consuming time and developmentLight PCR method and biochip technology. In the time that the kind of carrying out species with mtdna sequence is differentiated, conventional region comprisesThe Cytochromeb gene (cytB) of Codocyte pigment B, small subunit ribosome 12SRNA coded sequence (12SRibosomalRNA), cromoci subunit I(cytochromeCoxidasesubunitI) encoding gene coxI,Atp8 and atp6 gene and the control zone D-loop fragment of coding ATP8 and ATP6. The sequence variations in these regions is moderate,In planting, there is certain conservative, embody again certain interspecific difference.
Fluorescent quantitative PCR technique is that a kind of highly sensitive nucleic acid detects and quantitative approach, and general principle is at real-time fluorescenceIn quantitative PCR reaction, introduced a kind of fluorescence chemical material, along with the carrying out of PCR reaction, PCR product constantly accumulates, glimmeringLight signal strength also equal proportion increases. Fluorescent quantitative PCR technique steps qualitative detection and has gone up the step that can quantize, and hasHighly sensitive, specificity and reliability be stronger, can realize multiple reaction, automaticity is high, nonstaining property, have real-time andThe features such as accuracy, particularly rely on the TaqMan method of probe combination, be widely used at present molecular biology research andThe fields such as medical research. In species qualifications application aspect, the domestic national standard of having issued or professional standard contained goose,Mammal or the aquatiles such as sheep, deer, camel, horse, donkey, pig, rabbit, dog and fish. In view of the importance of ruminant, invention speciallyProfit " fluorescence PCR detection reagent and the preparation method and application of discriminating ruminant derived component " (publication number: CN101659996A) describe a kind of fluorescence PCR method that can simultaneously differentiate ox, sheep and goat, carried out animal with three probesScreen in source, patent of invention that publication number is CN102311999A " donkey or donkey derived component real-time fluorescence PCR detection method andDetect with primer and probe " provide with particular probe and differentiated that donkey carrys out the fluorescence PCR method of derived components, and publication number is CNThe patent of invention " horse or horse derived component real-time fluorescence PCR detection method and primer and probe for detection " of 102311998AA kind of fluorescence PCR method of identifying Malaysia derived components is provided. Publication number is patent of invention " a kind of animal of CN101196463The discrimination method of derived component " provide one group can differentiate vertebrate primer sequence and the routines such as ox, sheep, pig, horse, goosePCR-gel electrophoresis method, similarly, aspect the qualification of poultry source, " a pair of goose derived component PCR detects patent of inventionWith primer " (publication number: CN102337337A) provide a pair of drawing for conventional pcr analysis E Xianli D-loop district's fragmentThing and agarose gel electrophoresis authentication method, patent of invention that publication number is CN102382897 " goose composition in food and feedsReal-time fluorescence PCR detection method and kit " disclose one and utilize TaqmanMGB probe with real time quantitative PCR method pairThe method that goose mitochondria 12SRNA fragment increases and analyzes for improving the efficiency detecting, can be examined multiple bird simultaneouslySurvey, " PCR-mtDNA detect the amplimer of total avian composition and detection kit thereof and using method (publication number:CN101270392) " method that detects multiple poultry derived mitochondrial DNA with pair of primers, said method are just further providedIn, real-time quantitative PCR detects strong to device dependence, is difficult to realize Site Detection, and conventional pcr amplification detects need to be to productAfter gel electrophoresis, carry out IMAQ and analysis, expend time in longer, and be difficult to ensure the sensitivity of detection.
Detect the detection effect that can effectively improve nucleic acid product by the method for immunology detection being transplanted in nucleic acid productRate. Can in its amplified production, introduce simultaneously by the specific protein of primer mark or compound that pcr amplification is usedComprise the nucleic acid complexes of two kinds of compound/protein labelings, the product of this mark can be by antibody or part with classMode like double-antibody sandwich detects, the immuno-gold labeling detection technique of the similar routine of this process, and this method isIn the method for SNP and constant-temperature amplification target gene, use, particularly to pathogenic microorganism or disease association baseThe qualification of cause, patent of invention-one that publication number is CN102134596A is based on nucleic acid cross-current ELISA test strip monokaryon glycosidesThe method of acid polymorphism carries out the detection of SNP with the method. Send out and publication number is CN102520172ABright patent-mono-increasing listeria spp nucleic acid chromatography detection kit and detection method thereof and application have been described one and have been adopted respectively lifeSingle listeria spp genome detection method that increases of thing element and digoxigenin labeled primer. Therefore only having amplified production of class methodsWithout judging, the molecular weight of product is not confirmed, therefore the disturbing factor in amplified reaction is more more, and common causesFalse-positive reason comprises: the abnormal renaturation between non-specific amplification, primer dimer, amplified production, solution primer dimer,The interference that abnormal annealing causes can be from the control of the selection of PCR polymerase, primer sequence optimization, dNTP substrate, crossover processCarry out several aspects. The archaeal dna polymerase that selection has thermal starting (Hot-start) function can obviously reduce primer dimer andThe formation of non-specific amplification product. Aspect primer optimization, patent of invention-" a pair of portion that publication number is CN102146432APoint reduce its dimeric method with order primer " described a kind of at primer 5 ' end the design of primers side with short palindromic sequenceMethod, recirculation under this primer normal temperature, avoids forming heterodimer, patent of invention that publication number is CN102719547A-" detecting the real-time fluorescence quantitative PCR reagent of HER2 gene expression dose " has also adopted similar method for real-time quantitative PCRAmplification; Publication number is that patent of invention-" using chimeric primers to reduce heterodimer forms " of CN101842494A describedA kind of method that uses chimeric primers to increase; In to the optimization of reaction substrate, publication number CN101171343A sends outBright patent " 3 ' modified oligonucleotide that contains false iso-cytosine nucleobase derivative and the application as primer or probe thereof "A kind of method that provides nucleotides that uses special modification to form to reduce primer dimer as substrate, use probe orIntroduce the interference that internal control probe also can reduce non-specific amplification, patent of invention-" one that publication number is CN101957373AAdd internal control nucleic acid pathogen nucleic acid to be carried out to the method for half-quantitative detection " adopt internal control probe to reduce interference, above-mentioned sideIn method, using thermal starting technology and cyclisation primer is current method, all the other methods or adopted special synthetic substrate andSubstrate, or need to introduce crossover process for the second time by probe, all can increase the complexity of testing process, particularly probeHybridizing method, because needing insulating process to lose the convenience of test strips method.
In design of primers of the present invention, keeping on specificity basis, in primer pair and primer 5 ' and 3 ' end annealingDimeric formation is avoided in the region that it is suitable that free energy is assessed rear selection, for the dimeric interference of ELISA test strip methodSituation is with suitable amplicon length, and in cooperation expansion buffer solution, the concentration optimization of detergent and denaturant, can realize primerEffective removing of dimer and non-specific amplification product.
Summary of the invention
Can be by the antibody specificity of this little molecule or compound by the nucleic acid molecules of biological micromolecule or compound markIdentification, thus detect by immunological method. The common molecule that can be used for labeled nucleic acid molecule comprises hapten molecule biologyElement (Biotin), digoxin (Digoxin), luminescent dye molecule rhodamine (RBITC), fluorescein isothiocynate(fluoresceinisothiocyanate, FITC), Cy3, Cy5 etc. In above-mentioned labelled molecule, digoxin, isothiocyanic acid are glimmeringLight element is all easy to obtain special antibody, and biotin molecule and its part-streptavidin have the binding characteristic of high special,Therefore these several molecules can be selected mark and the immunology detection for nucleic acid molecules. The present invention is intended to antigen or half to resistThe few nucleic acid of former little molecular labeling strand is as primer, and to target nucleic acid specific amplification to be checked, the complex molecule of formation simultaneouslyPossess two kinds of antigens or hapten-marked, immunogenic, this compound is to be availablely combined with specific antibody or sepcific ligands, realThe immune colloid gold of existing nucleic acid amplification product detects.
Therefore, on the one hand, the invention provides a kind of test strips method detect goose derived components in food and feeds detection skillArt, mainly comprises:
1) two unique amplimers of design, wherein forward primer Geese-12s-F has following sequence: 5 '-TTAKAGCAACAGCCTAACTTCA-3 ', with antigen or hapten molecule biotin (Biotin), fluorescein isothiocynate(FITC) a kind of mark or in digoxin (Digoxin); The sequence of downstream primer Geese-12s-R is 5 '--CTAAGATCATCTTAATGGTGTG-3 ', to be different from a kind of mark of upstream primer; Under suitable amplification condition, can expandIncrease the DNA fragmentation that goose mitochondrial genomes 12s region one segment length is 196bps; When without tested nucleic acid-templated existence, without specialOpposite sex nucleic acid fragment amplified production;
2) by a kind of universal antibody of standard, as crosslinked in anti-FITC mAb and colloid gold particle, form bag at particle surfaceThe antibody of quilt;
3) by the antibody of another kind of labelled molecule or part, as part-chain parent of anti digoxin antibody or biotin moleculeBe fixed on and on film, form detection line with linear with plain molecule;
4) in the time there is specific amplification products to be checked, because of the effect of upstream and downstream primer, in amplified production simultaneously withIn above-mentioned three kinds of labels two kinds, form the compound of labelled molecule 1-amplified production-labelled molecule 2;
5) by step 4) the mark 1-amplified production-mark 2 forming and the mark 1 of colloid gold particle pan coatingAntibody combination, form anti-mark 1 antibody-mark 1-amplified production-mark 2 coloured particle compounds;
6) by step 5) the coloured particle compound that obtains travels up to along tunica fibrosa by capillarity in solutionBe coated with on the antibody of mark 2 or the lines of part, compound is because depositing with antibody (part) combination of mark 2, stagnantStay on detection line, form macroscopic colored line, be judged to be the positive; 7) in the time that specific amplification products does not exist, noThere is above-mentioned steps 4)-6), can not form mark 1-amplified production-mark 2 compounds, also just can not shape be deposited on detectionOn line, on the antibody of mark 2 (part), do not form visible band, be judged to feminine gender.
On the other hand, the present invention also provides the developping solution detecting for nucleotide sequence, and this developping solution can reduce primerThe interference of dimer to experimental result;
On the other hand, the invention provides the test strips for the fast detecting of food and feeds goose derived components, its bagDraw together at the liner with adhesive sticker, have in order: 1: absorbent filter pad; 2: bond pad, with the first nucleic acid markersThe particle that adds lustre to (collaurum or latex) of antibody or part; 3: lateral chromatography matrix (nitrocellulose filter or nylon membrane); 4: inspectionMeasuring tape, with the antibody of the second nucleic acid markers; 5: quality control band, having can be in conjunction with the antibody of specific Species origin antibody; 6:Substrate; 7: adsorptive pads.
On the other hand, the present invention also provides the universal standard test strips for detection of nucleic acid amplification product.
Finally, the present invention also provide above-mentioned detection method and test strips in food and feeds goose derived component detect inPurposes.
The explanation of technical terms using in description of the present invention:
The starting material that primer: DNA is synthetic. Be generally a pair of strand oligonucleotide, with template hybridization after, DNA synthetic fromIts 3 ' end starts.
Mark: by detectable signaling molecule (as haptens, fluorescence, radioactivity etc.) and the coupling of single stranded oligonucleotide phaseMethod.
Hybridization: refer in particular to complementary DNA single chain and form duplex structure by base pairing.
Launch: opened by test strips adsorptive pads bottom under the chromatography effect of expansion buffer solution through the PCR of amplified reaction productThe process moving to detection line, nature controlling line direction begins.
Nucleic acid: the common name of DNA (DNA) and ribonucleic acid (RNA).
Antigen and haptens: possess immunogenic material. Be generally macro-molecular protein or cellular component. But some is littleMolecule also possesses immunogenicity, is called as haptens ((Hapten). Haptens is often used to label probe.
Antibody: the protein molecule that can be combined with antigen or hapten specificity.
Complex: the bond of being formed by two or more molecular specificity.
Primer dimer: in the process of PCR (PCR), because of between primer pair or wall scroll primer mutually annealThe dimer molecule forming, the dimer being made up of two different primers is heterodimer, because drawing with two kinds of marksRise with planting the combination of antibody (part) and cause false positive, the dimer being formed by single primer annealing is homodimer, only bandHave a kind of mark and can not cause false positive, but the meeting of formation of too much dimer reduces amplification efficiency.
Immunity test strip: for the medical tool of fast detecting. Be called again colour generation membrane chromatographic.
Nucleic acid polymerase: the enzyme of nucleic acid long-chain. Be divided into archaeal dna polymerase and the large class of RNA polymerase two.
Beneficial effect
Because the present invention is the genetic fragment with the primer amplified suitable length of mark, can ensure the height detectingSpend specificity, thereby ensured the accuracy of testing result. Novel part of the present invention has been avoided amplified production dilution and probeThe flow processs such as hybridization, have kept the advantages such as immune colloid gold (test strips) technology is directly perceived, quick, convenient, ripe, cheap, possess againThe feature of pcr amplification method height sensitivity, high degree of specificity, therefore, method of the present invention is a kind of effectively goose source animalComponent detection method.
The current techique platform detecting as a nucleic acid amplification product, method of the present invention and test strips thereof can be extensiveFor the qualification of feed animal sources composition and the detection of food borne pathogenic microorganism, have in farming and animal husbandry, customs inspection quarantineApplication prospect widely. Though have animal sources Components identification method, its design of primers and the product fragment of multiple PCR-based at presentAll be suitable for sonde method or gel electrophoresis, and for the interference that reduces primer dimer and non-specific renaturation in colloidal gold strip methodRemove and lack necessary consideration, solution is also also inapplicable. It is a kind of based on immune colloid gold side that the object of the invention is to provideMethod detects PCR primer, amplification and the detection method of goose source animal component, and the application of the method. Other common species, asThe qualification of sheep, pig, ox, donkey, rabbit, camel, sheep, chicken, duck, fox, ermine, deer etc. can adopt method similarly to complete.
The invention of nucleic acid test strip detection method is by the trace routine of greatly simplifying after nucleic acid amplification. Result interpretation is simply bright, directly perceived (detecting signal result as accompanying drawing 2).
Goose derived components test strips Fast Detection Technique in the food and feeds that this patent is applied for, as a kind of novel coreDetection technique after acid amplification, has the following advantages:
Simple to operate: only the sample after nucleic acid amplification directly need to be dripped to nucleic acid reagent and detect version above, not needProfessional's operation, is convenient to promote;
Quick: to detect sentence read result after 5 minutes;
Sensitive: compared with agarose gel electrophoresis, detection sensitivity improves nearly 100～200 times;
Specificity is high: be specificity labeled primers due to what add use in testing process, make result more accurate;
Low price: expense is significantly less than traditional gel electrophoresis and ELISA detects.
Brief description of the drawings
Fig. 1 shows the basic structure of the test strips of amplification of nucleic acid sequences product detection of the present invention;
Fig. 2 is the testing result schematic diagram of detection of nucleic acids test strips;
Fig. 3 is the method showing with embodiment 1, adopts specificity labeled primers to detect the inspection of goose composition PCR-test stripsSurvey result;
Fig. 4 is the method showing with embodiment 2, the specific PCR-test strips of employing detection goose specificity labeled primersTesting result;
Fig. 5 is the method with embodiment 2, the susceptibility comparison of test strips and detected through gel electrophoresis method after show nucleic acid amplificationResult.
Detailed description of the invention
Following embodiment illustrates detection method of the present invention and detects effect. Embodiment is only used for for example, noForm any restriction to protection domain of the present invention, protection scope of the present invention illustrates at appending claims.
Embodiment 1
1 materials and methods
Goose mitochondrial DNA
1.2 design of primers
1.3PCR amplification system:
1.3PCR amplification system:
Reaction condition:
Get respectively 5 μ l simultaneously and examine for nucleic acid test strip, measure 5 μ L amplified productions, join in 95 μ L developping solutionsRow test point is in sample pad, observed result after 5 minutes.
The experiment of 1.4PCR specificity
The PCR reaction system that utilization is set up is to 1: mouse; 2: ox; 3: rabbit; 4: horse; 5: sheep; 6: dog; 7: pig; 8: cat; 9:Chicken; 10: duck; 11: goose; 12:NTC blank (water) is verified its specificity.
2 results
2.1PCR reaction system and condition
The HSTaqDNApolymerase that adopts TaKaRa company, overall reaction system is 20 μ l. Use Bio-RadPCRInstrument detects, and response parameter is: 94 DEG C of 5min, and 94 DEG C of 30s, 55 DEG C of 30s, 72 DEG C of 30s increase, and 30 are followedAfter ring, proceed to 72 DEG C of 5min, 4 DEG C of preservations. Get 5 μ l products containing carrying out electricity in 2% agarose gel electrophoresis of ethidium bromideSwimming, after electrophoresis, judgement has or not specific band. After nucleic acid amplification, the testing result of test strips and gel electrophoresis is shown in accompanying drawing 4In.
2.2 specific test
The PCR method that the present invention sets up has good specificity to goose genomic DNA, only has slight friendship with rabbit mitochondriaFork, can be diluted and be avoided by sample.
Embodiment 2
The sensitivity of nucleic acid test strip detection method, by pcr template with 1/10,1/102，1/103，1/104，1/105，1/106，1/107，1/108，1/109，1/1010Dilution proportion after, increase by the PCR system of having set up.
1 materials and methods
Goose mitochondrial DNA
1.2 design of primers
1.3PCR amplification system:
Reaction condition:
Get respectively 5 μ l for agarose gel electrophoresis. Gel electrophoresis condition is: 1XTBE buffer solution, voltage 100V, electricity30 points of swimming times. Get respectively 5 μ l simultaneously and detect for nucleic acid test strip, get 5 μ l sample spot in sample pad, join 95 μ LIn developping solution, detect observed result after 5 minutes.
2 results
2.1PCR reaction system and condition
The HSTaqDNApolymerase that adopts TaKaRa company, overall reaction system is 20 μ l. Use Bio-RadPCRInstrument detects, and response parameter is: 94 DEG C of 5min, and 94 DEG C of 30s, 56.5 DEG C of 30s, 72 DEG C of 80s increase, 30After circulation, proceed to 72 DEG C of 5min, 4 DEG C of preservations. Get 5 μ l products containing carrying out electricity in 2% agarose gel electrophoresis of ethidium bromideSwimming, after electrophoresis, judgement has or not specific band. The susceptibility comparative result of test strips and gel electrophoresis after nucleic acid amplification is shown inIn accompanying drawing 5, can be found out by the result of accompanying drawing 5, compared with traditional gel electrophoresis, its susceptibility increases approximately 100 times.
TTAGAGCAACAGCCTAACTTCAAGGGTTGGTAAATCTTGTGCCAGCCACCGCGGTCATACAAGAAACCCAAATCAACCGTCCTGTTGACACGGCGTAAAGAGTGGTAAAATGCCTATCCTAGCTAACTAAGATCAAAATGCAACTGAGCTGTCATAAGCCCAAGATGCACCTAAACACACCATTAAGATGATCTTAG
Claims (3)
1. a PCR-immunity colloidal gold test paper strip method for quick for goose derived components in food and feeds, it is upper that PCR usesTrip primer is: Geese-12s-F:5 '-TTAGAGCAACAGCCTAACTTCA-3 ', and downstream primer is: Geese-12s-R:5 '-CTAAGATCATCTTAATGGTGTG-3 ', selects biotin molecule Biotin, fluorescein isothiocynate FITC or digoxinIn Digoxin, two kinds of different marks, 5 ' end to primer carries out mark respectively so that detect, and the mark of upstream and downstream primerDifferent; The amplification condition of PCR is: 95 DEG C of denaturation 5min, 94 DEG C of sex change 30sec; 55 DEG C of annealing 30sec; 72 DEG CExtend 30sec, 30 circulations of increasing, last 72 DEG C are extended 5min.
2. detection method as claimed in claim 1, is characterized in that, adopts composition to be: 10mMTris, 1%BSA, 1%The developping solution of the NaOH of Tween20 and 0.05mol/L launches colour developing to amplified production.
3. detection method as claimed in claim 1, is characterized in that, also comprises and adopts test strips to carry out nucleic acid amplification productDetect step, the coloured particle of described test strips is colloid gold particle, the matrix of described test strips be nitrocellulose filter orNylon membrane.
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| CN104017872B (en) * | 2014-05-29 | 2017-02-08 | 天津出入境检验检疫局动植物与食品检测中心 | Lateral flow test paper strip detection kit for detection of fox source components in food and feed and application thereof |
| CN109852708B (en) * | 2019-04-11 | 2021-01-01 | 中国农业大学 | Kit for rapidly identifying goose-derived components in food and application thereof |
| CN109811065B (en) * | 2019-04-11 | 2021-04-06 | 中国农业大学 | Rapid detection kit for goose-derived components in food and application thereof |
| CN109825611B (en) * | 2019-04-11 | 2020-11-24 | 中国农业大学 | Method and kit for rapidly detecting goose-derived components in food |
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| CN101196463A (en) * | 2007-12-14 | 2008-06-11 | 东北农业大学 | Identification method for animal derived materials |
| CN102337337A (en) * | 2011-09-28 | 2012-02-01 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Polymerase chain reaction (PCR) detection primer for goose-derived component |
| CN102382897A (en) * | 2011-12-07 | 2012-03-21 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | Real-time fluorescent PCR (Polymerase Chain Reaction) detection method and kit for goose components in food and feedstuff |
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2014
- 2014-03-05 CN CN201310670537.4A patent/CN103773846B/en not_active Expired - Fee Related
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| CN101196463A (en) * | 2007-12-14 | 2008-06-11 | 东北农业大学 | Identification method for animal derived materials |
| CN102337337A (en) * | 2011-09-28 | 2012-02-01 | 深圳出入境检验检疫局动植物检验检疫技术中心 | Polymerase chain reaction (PCR) detection primer for goose-derived component |
| CN102382897A (en) * | 2011-12-07 | 2012-03-21 | 上海出入境检验检疫局动植物与食品检验检疫技术中心 | Real-time fluorescent PCR (Polymerase Chain Reaction) detection method and kit for goose components in food and feedstuff |
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