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CN105891479A - Stomach disease detection kit - Google Patents

Stomach disease detection kit Download PDF

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Publication number
CN105891479A
CN105891479A CN201610504405.8A CN201610504405A CN105891479A CN 105891479 A CN105891479 A CN 105891479A CN 201610504405 A CN201610504405 A CN 201610504405A CN 105891479 A CN105891479 A CN 105891479A
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China
Prior art keywords
detection kit
disease detection
nucleic acid
stomach disease
aptamer
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CN201610504405.8A
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Chinese (zh)
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卢美珍
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Individual
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    • 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
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/16—Aptamers
    • 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
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)
    • G01N2333/95—Proteinases, i.e. endopeptidases (3.4.21-3.4.99)
    • G01N2333/964—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue
    • G01N2333/96425—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals
    • G01N2333/96427—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general
    • G01N2333/9643—Proteinases, i.e. endopeptidases (3.4.21-3.4.99) derived from animal tissue from mammals in general with EC number
    • G01N2333/96472—Aspartic endopeptidases (3.4.23)
    • G01N2333/96475—Aspartic endopeptidases (3.4.23) with definite EC number
    • G01N2333/96477—Pepsin (3.4.23.1; 3.4.23.2; 3.4.23.3)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/06—Gastro-intestinal diseases
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/06—Gastro-intestinal diseases
    • G01N2800/062—Gastritis or peptic ulcer disease

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • General Engineering & Computer Science (AREA)
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  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Hematology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • Biophysics (AREA)
  • Pathology (AREA)
  • Plant Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention relates to a stomach disease detection kit, and belongs to the technical field of in-vitro reagent diagnosis. The stomach disease detection kit is characterized in that a nucleic acid aptamer which can be specifically combined with pepsinogen II is screened by a SELEX (systematic evolution of ligands by exponential enrichment) technique, the pepsinogen II in blood is specifically screened or detected by the nucleic acid aptamer, and the nucleic acid aptamer is prepared into the detection kit, so that the direct reaction for detection without thinning of blood is realized, and the workload is decreased; the measuring range is wide, and the stomach disease detection kit is suitable for clinical application. The stomach disease detection kit has the beneficial effects that the sensitivity is high, the specificity is good, the measuring range is wide, the operation is simple, the stomach disease detection kit is suitable for large-scale popularization and application, and the market prospect is broad.

Description

A kind of test kit for detection of stomach disorders
The application is filing date on November 29th, 2015, Application No. 201510849189.6, invention entitled " a kind of Test kit for detection of stomach disorders " the divisional application of application for a patent for invention.
Technical field
The application relates to field of biological detection, is specifically related to the detection of gastropathy.
Background technology
Pepsinogen I I (being called for short PG II) derives from full gastric gland and distal duodenum BrunnerShi gland, and gastric mucosa closes The PGII become is about the 25% of total amount, and PG II major part enters digestive tract, enters blood circulation on a small quantity, is therefore referred to as " reflection Stomach state and the pointer of function ".
Clinical research both at home and abroad is pointed out, PG II is relatively big with the dependency of gastric mucosa pathological changes, and its rising is withered with fundic gland pipe Contracting, intestinal epithelial metaplasia or Pseudopyloric gland metaplasia, dysplasia are relevant.Measure PG II content help detect duodenal ulcer, Other digestive tract disease such as atrophic gastritis, gastritis, gastric cancer, for Clinical detection and health check-up examination demand.In crowd's gastropathy examination In, everyone makees gastroscope is unpractical, can be detected superficial gastritis, erosive gastritis, stomach by Noninvasive serum PG Out, then to carry out gastroscopy be a kind of realistic plan to the high-risk Mass screenings such as ulcer, duodenal ulcer, gastric cancer. Employing time-resolved fluorescence known in the art measures technology, and the TRFIIA test kit of the PGII of establishment is current PGII detection side In method the sensitiveest, measure one of the method for widest range, but expensive be unfavorable for large-scale promotion.
Phyletic evolution index concentration technology (SELEX technology) is a kind of new combination grown up early 1990s Chemical technology, it uses jumbo random oligonucleotide library, the outer PCR amplification technique of coalition, divides with target with index concentration The oligonucleotide of sub-specific bond, through multi-turns screen, it is thus achieved that affinity is high, the oligonucleotide aptamer of high specificity (aptamers).The own Successful utilization of this technology is in the screening of many target molecules, including metal ion, organic dyestuff, medicine, albumen Matter, aminoacid and various cytokines etc..
Summary of the invention
It is an object of the invention to the widow by phyletic evolution index concentration technology (SELEX technology) screening pepsinogen I I Nucleotide aptamer substitutes antibody, it is provided that a kind of succinct quick, pepsinogen I I inspection in early days of high sensitivity, high specific Survey and isolation and purification method.
Technical scheme:
For single stranded DNA random library and the primer of phyletic evolution index concentration technology screening in the present invention, by the U.S. Invitrogen company synthesizes, and two ends are fixed sequence program, and centre is the random sequence of 42 bases: 5'- TACGACATGAACCGTGATAA (N42) CAGTGAAACCTGATGATCGA-3', storage capacity is 1014Above；
Primer 1:5'TACGACATGAACCGTGATAA-3'；
Primer 2: 5'-TCGATCAGCAGGTTTCACTG-3'.
Human pepsinogen II is fitted together to egg according to the recombined human pepsinogen I I isozyme disclosed in CN103387971A White method prepares；
GelRed nucleic acid dye is purchased from Biotium company；
Nitrocellulose filter is purchased from U.S. Mi Libo (MilliPore) company；
It is Time Inc. that the purified reagent of oligonucleotide is purchased from sky, Beijing；
PCR kit and carrier T are purchased from U.S. Pu Luomaige (ftOmega) company.
A kind of affine human pepsinogen II nucleic acid aptamer, it is characterised in that nucleotides sequence is classified as: SEQ ID NO:1- 17 arbitrary shown DNA moleculars.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: there is the oligonucleoside of identical function Acid aptamer homologous sequence accounts for more than 60%.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: derivative RNA sequence has identical Function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: for hybridizing with DNA sequence Oligonucleotide sequence.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: in any position of nucleic acid aptamer Oligonucleotide aptamer sequence obtained by putting deletion or increasing part oligonucleotide residues has identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: in any position of nucleic acid aptamer Putting after carrying out the displacement of nucleotide kind and rare bases, the oligonucleotide aptamer sequence obtained has identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: in any position of nucleic acid aptamer Put carry out phosphorylation, methylate, amino, sulfydryl, isotope, biotin, digoxin, fluorescent material, nano luminescent material or enzyme After labelling is modified, the oligonucleotide aptamer sequence obtained has identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: for human pepsinogen II's Detection is with isolated and purified, thus is used for the detection of disease of brain.
The preparation method of a kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, sequentially includes the following steps: 1) strand The synthesis of DNA random oligonucleotide library；2) utilize phyletic evolution index concentration method that oligonucleotide library is screened；3) Amplification and the oligonucleotide of human albumin specific bond；4) carry out next round screening, after 12 take turns above screening, obtain purpose Oligonucleotide sequence；5) cloning and sequencing.
The invention have the advantage that have simplicity, quickly, economic dispatch feature, with other combinatorial chemical libraries such as random peptide library, resist Body storehouse is compared with phage display libraries, the many advantages of aptamer tool filtered out from oligonucleotide library: 1) itself It is oligonucleotide, molecular weight, can be cost-effective with chemosynthesis；2) there is affinity more higher than antibody and specificity； 3) labelling and can be at different parts selectively labelling it is easy to；4) repeatability and good stability, and be prone to preserve, i.e. to height Gentle drastic conditions is insensitive.Therefore, phyletic evolution index concentration technology has a good application prospect.
Detailed description of the invention
The preparation of embodiment 1 human pepsinogen II
Method preparation according to the recombined human pepsinogen I I isozyme chimeric protein disclosed in CN103387971 A obtains Obtaining human pepsinogen II, protein concentration is 100mg/mL.
Embodiment 2 synthesizes random single-stranded DNA banks and primer
Random single chain DNA (ssDNA) library: 5'-TACGACATGAACCGTGATAA (N42) CAGTGAAACCTGATGATCGA-3', constructs the ssDNA pool of a length of 82nt, and two ends are immobilized primer sequence, in Between be the random sequence of 42 bases, storage capacity is I014Above；Primer 1:5'TACGACATGAACCGTGATAA-3'；Primer 2:5'-TCGATCAGCAGGTTTCACTG-3'；SsDNA pool and two kinds of primers are all become 100 μ with TE buffer The storage of mol/L stock solution _ 20 DEG C is standby.
Being double-stranded DNA by single-stranded DNA banks amplification, product is through 2% agarose gel electrophoresis and cuts glue recovery purification；To return The double-stranded DNA received is template, and in vitro transcription goes out single stranded RNA random library, and transcription product is through PAGE purification.75 μ g RNA library warps The anti-sieve of nitrocellulose filter removes the RNA molecule being combined with film, then with 2ug human pepsinogen II albumen, hatches for 37 DEG C 30min, reactant liquor filters through nitrocellulose filter, washs filter membrane；Then filter membrane is shredded, be placed in elution buffer (6mol/L Carbamide, 0.55mol/L ammonium acetate, l.5mmol/L EDTA, 0.15%SDS) in boil 5min, centrifugal, take supernatant, dehydrated alcohol Precipitation RNA, and be redissolved in 20 μ 1DEPC water；With RNA for template RT-PCR amplifying doulbe-chain DNA, in vitro transcription goes out RNA literary composition Storehouse is screened for next round；Often wheel screening process in RT-PCR obtain double-stranded DNA library, with this double-stranded DNA as template body outside turn Recording out RNA aptamer storehouse, screening carries out 10 altogether and takes turns.Having obtained 14 aptamers, its sequence is respectively SEQ ID NO:1-14 institute Show.Particular sequence is as follows:
PGII-1:
TACGACATGAACCGTGATAACTCATATATGTTCCCAACACGTCCAACTTATCCCTGACAATA
CAGTGAAACCTGATGATCGA
PGII-2:
TACGACATGAACCGTGATAACCGCTTCACACACTACCGCTCTCTCTTAGACTATTAACAATT
CAGTGAAACCTGATGATCGA
PGII-3:
TACGACATGAACCGTGATAAATTCATACTTGTACAAATACTCTCGCATCTCCACTTATAATA
CAGTGAAACCTGATGATCGA
PGII-4:
TACGACATGAACCGTGATAAATAATTCTTTGCTTACCCTAAATATTCCAAACCCATCGACCA
CAGTGAAACCTGATGATCGA
PGII-5:
TACGACATGAACCGTGATAAACCTTATTATCCTACCACATCTCCTCAAACCAGCATTTAATA
CAGTGAAACCTGATGATCGA
PGII-6:
TACGACATGAACCGTGATAACAATCCACTGCCTCATTCCTTCCTTTATTCACATATTCCAAA
CAGTGAAACCTGATGATCGA
PGII-7:
TACGACATGAACCGTGATAACGCTTATCTTCTCACATAATTCCGAAACATAAAACCTCTCCA
CAGTGAAACCTGATGATCGA
PGII-8:
TACGACATGAACCGTGATAAATCGCCAACACCCTCCGTCTCGCTCTCCTAATATACAATCCT
CAGTGAAACCTGATGATCGA
PGII-9:
TACGACATGAACCGTGATAATCGAACATTAACAATACCACGCCTATATTCTTAACATAAATA
CAGTGAAACCTGATGATCGA
PGII-10:
TACGACATGAACCGTGATAACACTTATAACAAACTCCAAGCCTCCTACAACGTACTATCACA
CAGTGAAACCTGATGATCGA
PGII-11:
TACGACATGAACCGTGATAAAAATATAAAGCTCTATGTTATTCACACGCATACTTCTTATCA
CAGTGAAACCTGATGATCGA
PGII-12:
TACGACATGAACCGTGATAACGCCCTACAATTCCCAATTAGCCTATTATTTAACATCCTAAT
CAGTGAAACCTGATGATCGA
PGII-13:
TACGACATGAACCGTGATAACCATATCTTGACCTATCACTTAGCCTAAATACTTTAAACATT
CAGTGAAACCTGATGATCGA
PGII-14:
TACGACATGAACCGTGATAACTACCTTCATACAATCGCAATATTCACTTTTAAACACAATAA
CAGTGAAACCTGATGATCGA
The performance measurement of embodiment 3 protein binding aptamer
Aptamer taking 2.0 μ g respectively, digests lh with calf intestinal alkaline phosphatase (CIP) 37 DEG C, purification reclaims and removes phosphoric acid The RNA changed；By T4 polynucleotide kinase labelling [γ-32P] ATP in dephosphorylized RNA molecule end.10nmol radioactivity The aptamer of labelling human pepsinogen II 37 DEG C with variable concentrations (1-200nM) respectively hatches 30min, respectively organizes reactant liquor Filtering through nitrocellulose filter, wash filter membrane, be dried filter membrane, liquid scintillation counter measures the exit dose of residual on filter membrane, with equally Product are parallel does twice mensuration.Calculate the dissociation constant of each aptamer and destination protein.Result is as follows:
Aptamer specificity analyses and stability analysis described in embodiment 4
It is respectively adopted human albumin, immune globulin, pg120 albumen, escherichia coli outer membrane protein A, pepsin Proenzyme II, carries out specific detection with 14 aptamers, through binding tests find, these aptamers the most not with these albumen phases In conjunction with, and only keep higher specificity with people's protein binding.
By described aptamer, take 0.2ug, be respectively placed in the serum of room temperature, aqueous solution, place surrounding.Pass through RT- PCR detects, and finds its Stability Analysis of Structures of placement of surrounding, is not degraded.
The diagnosis of aptamer disease described in embodiment 5
Take 9 patients w ith peptic ulcer diseases and the blood of 4 normal persons, use normal saline dilution, it is thus achieved that target sample.
By 14 markd aptamers of coupling respectively with the sample mixing 40min of 9 patients and 4 normal persons, logical Cross biotin to separate, the content of quantitative analysis human pepsinogen therein II, found by analysis, pepsin in 9 patients The content of proenzyme II dramatically increases, and has exceeded the threshold value of regulation.Reach the diagnostic criteria that corresponding gastropathy is sick.As can be seen here, its Diagnosis effect is preferable.
These are only the preferred embodiments of the present invention, be not limited to the present invention, for those skilled in the art For Yuan, all any modification, equivalent substitution and improvement etc. done within the spirit and principles in the present invention, should be included in this Within the protection domain of invention.
Sequence table
＜ 110 ＞ Lu Meizhen
＜ 120 ＞ mono-kind is for the test kit of detection of stomach disorders
〈160〉14
〈210〉1
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-1
TACGACATGAACCGTGATAACTCATATATGTTCCCAACACGTCCAACTTATCCCTGACAATACAGTGAAACCTGATGATCGA
〈210〉2
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-2
TACGACATGAACCGTGATAACCGCTTCACACACTACCGCTCTCTCTTAGACTATTAACAATTCAGTGAAACCTGATGATCGA
〈210〉3
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-3
TACGACATGAACCGTGATAAATTCATACTTGTACAAATACTCTCGCATCTCCACTTATAATACAGTGAAACCTGATGATCGA
〈210〉4
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-4
TACGACATGAACCGTGATAAATAATTCTTTGCTTACCCTAAATATTCCAAACCCATCGACCACAGTGAAACCTGATGATCGA
〈210〉5
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-5
TACGACATGAACCGTGATAAACCTTATTATCCTACCACATCTCCTCAAACCAGCATTTAATACAGTGAAACCTGATGATCGA
〈210〉6
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-6
TACGACATGAACCGTGATAACAATCCACTGCCTCATTCCTTCCTTTATTCACATATTCCAAACAGTGAAACCTGATGATCGA
〈210〉7
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-7
TACGACATGAACCGTGATAACGCTTATCTTCTCACATAATTCCGAAACATAAAACCTCTCCACAGTGAAACCTGATGATCGA
〈210〉8
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-8
TACGACATGAACCGTGATAAATCGCCAACACCCTCCGTCTCGCTCTCCTAATATACAATCCTCAGTGAAACCTGATGATCGA
〈210〉9
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-9
TACGACATGAACCGTGATAATCGAACATTAACAATACCACGCCTATATTCTTAACATAAATACAGTGAAACCTGATGATCGA
〈210〉10
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-10
TACGACATGAACCGTGATAACACTTATAACAAACTCCAAGCCTCCTACAACGTACTATCACACAGTGAAACCTGATGATCGA
〈210〉11
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-11
TACGACATGAACCGTGATAAAAATATAAAGCTCTATGTTATTCACACGCATACTTCTTATCACAGTGAAACCTGATGATCGA
〈210〉12
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-12
TACGACATGAACCGTGATAACGCCCTACAATTCCCAATTAGCCTATTATTTAACATCCTAATCAGTGAAACCTGATGATCGA
〈210〉13
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-13
TACGACATGAACCGTGATAACCATATCTTGACCTATCACTTAGCCTAAATACTTTAAACATTCAGTGAAACCTGATGATCGA
〈210〉14
〈211〉 82
〈212〉DNA
＜ 213 ＞ artificial sequence
〈400〉PG II-14
TACGACATGAACCGTGATAACTACCTTCATACAATCGCAATATTCACTTTTAAACACAATAACAGTGAAACCTGATGATCGA

Claims (3)

1. for the test kit of detection of stomach disorders, its contain can be specific binding with pepsinogen I I aptamer.
2. test kit as claimed in claim 1, it is characterised in that: described aptamer sequence is as shown in SEQ ID No:12.
3. the method for a detection of stomach disorders, it is characterised in that utilize the test kit described in any one of claim 1-2.
CN201610504405.8A 2015-11-29 2015-11-29 Stomach disease detection kit Pending CN105891479A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610504405.8A CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit

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Application Number Priority Date Filing Date Title
CN201610504405.8A CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201510849189.6A CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
CN201510849189.6A Division CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders

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Publication Number Publication Date
CN105891479A true CN105891479A (en) 2016-08-24

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Family Applications (14)

Application Number Title Priority Date Filing Date
CN201610504405.8A Pending CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610504404.3A Withdrawn CN106405083A (en) 2015-11-29 2015-11-29 A kit for stomach disease detection
CN201610505390.7A Pending CN105929161A (en) 2015-11-29 2015-11-29 Kit for detecting stomach diseases
CN201610503246.XA Pending CN106053809A (en) 2015-11-29 2015-11-29 Kit for detecting gastric disease
CN201610505427.6A Withdrawn CN106153916A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504439.7A Withdrawn CN106198977A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610497173.8A Withdrawn CN106153914A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504850.4A Pending CN105891480A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201510849189.6A Expired - Fee Related CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504436.3A Withdrawn CN106153915A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610503444.6A Pending CN105891478A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610503443.1A Withdrawn CN106198975A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504868.4A Pending CN105911280A (en) 2015-11-29 2015-11-29 Kit for stomach illness detection
CN201610504437.8A Pending CN105929160A (en) 2015-11-29 2015-11-29 Kit for stomach disease detection

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CN201610504404.3A Withdrawn CN106405083A (en) 2015-11-29 2015-11-29 A kit for stomach disease detection
CN201610505390.7A Pending CN105929161A (en) 2015-11-29 2015-11-29 Kit for detecting stomach diseases
CN201610503246.XA Pending CN106053809A (en) 2015-11-29 2015-11-29 Kit for detecting gastric disease
CN201610505427.6A Withdrawn CN106153916A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504439.7A Withdrawn CN106198977A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610497173.8A Withdrawn CN106153914A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504850.4A Pending CN105891480A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201510849189.6A Expired - Fee Related CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504436.3A Withdrawn CN106153915A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610503444.6A Pending CN105891478A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
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