CN106405083A - A kit for stomach disease detection - Google Patents

A kit for stomach disease detection Download PDF

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CN106405083A
CN106405083A CN201610504404.3A CN201610504404A CN106405083A CN 106405083 A CN106405083 A CN 106405083A CN 201610504404 A CN201610504404 A CN 201610504404A CN 106405083 A CN106405083 A CN 106405083A
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aptamer
kit
pepsinogen
dna
nucleic acid
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卢美珍
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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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  • Life Sciences & Earth Sciences (AREA)
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  • General Engineering & Computer Science (AREA)
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  • General Health & Medical Sciences (AREA)
  • Urology & Nephrology (AREA)
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  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention relates to a kit for stomach disease detection and belongs to the technical field of in-vitro agent diagnosis. A nucleic acid aptamer capable of specifically bonding pepsinogen II can be screened through an SELEX technique, and therefore the aptamer is utilized to specifically screen or detect the pepsinogen II in blood. Through preparing the aptamer into the kit, an objective of direct reaction detection without the need of blood dilution so as to reduce working loads is achieved, a wide measurement range is acquired, and clinical application is convenient. Beneficial effects of the kit are that the kit is high in sensitivity, good in specificity, wide in measurement range, and simple in operation, facilitates large-scale popularization and application and has a wide market prospect.

Description

A kind of test kit for detection of stomach disorders
The application is to be on November 29th, 2015, Application No. 201510849189.6, invention entitled " one kind the applying date The divisional application of the application for a patent for invention of the test kit for detection of stomach disorders ".
Technical field
The application is related to field of biological detection and in particular to the detection of gastropathy.
Background technology
Pepsinogen I I (abbreviation PG II) derives from full gastric gland and distal duodenum BrunnerShi gland, and gastric mucosa closes The PGII becoming is about 25%, PG II most of entrance digestive tract of total amount, 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 points out, PG II is larger 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 gastritiss, erosive gastritiss, stomach by Noninvasive serum PG Out, then to carry out gastroscopy be a kind of realistic plan to the high-risk Mass screening such as ulcer, duodenal ulcer, gastric cancer. Employing time-resolved fluorescence e measurement technology known in the art, the TRFIIA test kit of the PGII of establishment is current PGII detection side One of the sensitiveest in method, measurement range is the widest method, but expensive it is unfavorable for large-scale promotion.
Phyletic evolution index concentration technology (SELEX technology) is a kind of new combination growing up early 1990s Chemical technology, it uses jumbo random oligonucleotide library, in conjunction with external PCR amplification technique, is divided with target with index concentration The oligonucleotide of sub- specific bond, through multi-turns screen, obtains affinity height, the oligonucleotide aptamer of high specificity (aptamers).The own Successful utilization of this technology in the screening of many target molecules, including metal ion, organic dyestuff, medicine, albumen Matter, aminoacid and various cytokine etc..
Content of the invention
The purpose of the present invention is to screen the widow of pepsinogen I I by phyletic evolution index concentration technology (SELEX technology) Nucleotide aptamer, to substitute antibody, provides a kind of succinctly quick, high sensitivity, high specific pepsinogen I I early stage to examine Survey and isolation and purification method.
Technical scheme：
It is used for single stranded DNA random library and the primer of phyletic evolution index concentration technology screening, by the U.S. in the present invention Invitrogen company synthesizes, and two ends are fixed sequence program, the middle random sequences for 42 bases：5'- TACGACATGAACCGTGATAA (N42) CAGTGAAACCTGATGATCGA-3', storage capacity is 1014More than；
Primer 1:5'–TACGACATGAACCGTGATAA-3'；
Primer 2:5'-TCGATCAGCAGGTTTCACTG-3'.
Recombined human pepsinogen I I isozyme according to disclosed in CN103387971A for the human pepsinogen II is fitted together to egg 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 Beijing sky；
PCR kit and carrier T are purchased from U.S. Pu Luomaige (ftOmega) company.
A kind of affine human pepsinogen II nucleic acid aptamer is 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 Sour 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 being hybridized with DNA sequence Oligonucleotide sequence.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer it is characterised in that：Any position in nucleic acid aptamer Put the oligonucleotide aptamer sequence obtained by deleting or increasing part oligonucleotide residues and there is identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer it is characterised in that：Any position in nucleic acid aptamer Put after carrying out the displacement of nucleotide species and rare bases, the oligonucleotide aptamer sequence obtaining has identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer it is characterised in that：Any position in 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 obtaining 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 Detect and isolate and purify, thus the detection for disease of brain.
A kind of preparation method of above-mentioned affine human pepsinogen II nucleic acid aptamer, is carried out according to the following steps：1) single-stranded The synthesis of DNA random oligonucleotide library；2) using phyletic evolution index concentration method, oligonucleotide library is screened；3) Amplification and the oligonucleotide of human albumin specific bond；4) carry out next round screening, after more than 12 wheels screening, obtain purpose Oligonucleotide sequence；5) cloning and sequencing.
It is an advantage of the invention that：Have simplicity, quick, economical the features such as, with other combinatorial chemical libraries such as random peptide library, anti- Body storehouse is compared with phage display libraries, the many advantages of aptamer tool filtering out from oligonucleotide library：1) in itself It is oligonucleotide, molecular weight, can be cost-effective with chemosynthesis；2) there is the affinity higher than antibody and specificity； 3) it is easy to labelling and can be in different parts selectively labelling；4) repeatability and good stability, and be easy to preserve, that is, to height Gentle drastic conditions are insensitive.Therefore, phyletic evolution index concentration technology has a good application prospect.
Specific embodiment
The preparation of embodiment 1 human pepsinogen II
The method preparation of the recombined human pepsinogen I I isozyme chimeric protein according to disclosed in CN103387971A obtains Obtain human pepsinogen II, protein concentration is 100mg/mL.
Embodiment 2-in-1 one-tenth random single-stranded DNA banks and primer
Random single chain DNA (ssDNA) library:5'-TACGACATGAACCGTGATAA(N42) CAGTGAAACCTGATGATCGA-3', constructs the ssDNA pool that length is 82nt, and two ends are immobilized primer sequence, in Between for 42 bases random sequences, storage capacity be I014More than；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.
Single-stranded DNA banks are expanded as double-stranded DNA, product through 2% agarose gel electrophoresiies and cuts glue reclaim 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 Nitrocellulose filter is counter to be weeded out except the RNA molecule with film combination, then with 2ug human pepsinogen II albumen, 37 DEG C of incubations 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, centrifugation, 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 used for next round and screens；Often in wheel screening process, RT-PCR obtains double-stranded DNA library, is turned in vitro with this double-stranded DNA for template Record out RNA aptamer storehouse, screening carries out 10 wheels altogether.14 aptamers are obtained, 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 is taken respectively 2.0 μ g, 37 DEG C digest lh with calf intestinal alkaline phosphatase (CIP), and purification reclaims and removes phosphoric acid The RNA changing；By T4 polynucleotide kinase labelling [γ -32P] ATP in dephosphorylized RNA molecule end.10nmol radioactivity The aptamer of labelling 37 DEG C of incubation 30min of human pepsinogen II with variable concentrations (1-200nM) respectively, each group reactant liquor Through nitrocellulose filter filtration, wash filter membrane, filter membrane be dried, liquid scintillation counter measures the exit dose of residual on filter membrane, with Parallel the doing of product measures twice.Calculate the dissociation constant of each aptamer and destination protein.Result is as follows：
Aptamer specificity analyses and stability analyses 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 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, in aqueous solution, place surrounding.By RT- PCR detects, finds its Stability Analysis of Structures of placement of surrounding, is not degraded.
The diagnosis of aptamer disease described in embodiment 5
Take the blood of 9 patients w ith peptic ulcer diseases and 4 normal persons, using normal saline dilution, obtain target sample.
14 are coupled the markd aptamer sample mixing 40min with 9 patients and 4 normal persons respectively, lead to Cross biotin to separate, the content of quantitative analyses 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 of corresponding gastropathy disease.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 member, all any modification, equivalent substitution and improvement done within the spirit and principles in the present invention etc., should be included in this Within the protection domain of invention.
Sequence table
＜ 110 ＞ Lu Meizhen
A kind of test kit for detection of stomach disorders of ＜ 120 ＞
〈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. a kind of test kit for detection of stomach disorders, it contains can be with the aptamer of pepsinogen I I specific binding.
2. test kit as claimed in claim 1 it is characterised in that：Described aptamer sequence such as SEQ ID No：Shown in 14.
3. a kind of method of detection of stomach disorders is it is characterised in that utilize the test kit described in any one of claim 1-2.
CN201610504404.3A 2015-11-29 2015-11-29 A kit for stomach disease detection Withdrawn CN106405083A (en)

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CN201610504404.3A CN106405083A (en) 2015-11-29 2015-11-29 A kit for stomach disease detection
CN201510849189.6A CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders

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

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CN201610503444.6A Pending CN105891478A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610504868.4A Pending CN105911280A (en) 2015-11-29 2015-11-29 Kit for stomach illness detection
CN201610504850.4A Pending CN105891480A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610504436.3A Withdrawn CN106153915A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610503246.XA Pending CN106053809A (en) 2015-11-29 2015-11-29 Kit for detecting gastric disease

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CN201610504437.8A Pending CN105929160A (en) 2015-11-29 2015-11-29 Kit for stomach disease detection
CN201610504439.7A Withdrawn CN106198977A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610503443.1A Withdrawn CN106198975A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201510849189.6A Expired - Fee Related CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504405.8A Pending CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610505390.7A Pending CN105929161A (en) 2015-11-29 2015-11-29 Kit for detecting stomach diseases
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Family Cites Families (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004040306A1 (en) * 2002-10-31 2004-05-13 Labrea Holding S.A. Diagnostic device and kit
FI20065542A0 (en) * 2006-09-01 2006-09-01 Biohit Oyj Method and Sampling Kit for Assessment of Abdominal Mucosal Condition
RU2519646C2 (en) * 2007-10-26 2014-06-20 Биохит Оий Methods and products for diagnostics of autoimmune diseases and gastric cancer, associated with atrophic gastritis
US20130217046A1 (en) * 2010-06-23 2013-08-22 The Board of Trustees of the Univer. of Illinois Method for determining predisposition to esophageal-related disorders
CN102839181B (en) * 2011-06-23 2014-05-14 复旦大学附属华山医院 Nucleic acid aptamer sequence of gastric cancer cells, and uses thereof
CN102426236A (en) * 2011-08-31 2012-04-25 内蒙古科慧生物科技有限责任公司 Pepsinogen II (PGII) quantitative determination kit and detection method thereof
CN103305521B (en) * 2012-03-06 2016-06-08 复旦大学附属华山医院 The sequence of the aptamer of a kind of stomach cancer cell and application
CN102645537B (en) * 2012-04-26 2014-07-30 北京美康生物技术研究中心 Latex enhanced turbidimetric immunoassay kit for diagnosing gastric diseases or gastric cancer, preparation method thereof and application
CN103320445B (en) * 2013-07-11 2017-07-28 重庆市肿瘤研究所 The DNA aptamer GCA 5 of specific recognition stomach cancer cell and its application
CN104418937B (en) * 2013-08-22 2018-01-02 朱建安 People PGII epitope peptides, antigen, antibody, purposes and kit
CN103627701B (en) * 2013-12-04 2015-07-15 武汉大学 Screening of aptamers specially targeting gastric cancer cells and aptamers obtained after screening
CN104818278A (en) * 2015-04-17 2015-08-05 刘红卫 Aptamer capable of being in specific binding to TS/MDEP protein in gastric cancer cells
CN104830867A (en) * 2015-06-07 2015-08-12 杨洋 Aptamer capable of being specifically combined with DKK1 protein in cancer cells

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CN105891478A (en) 2016-08-24
CN106153914A (en) 2016-11-23
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CN105277698A (en) 2016-01-27
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CN105891479A (en) 2016-08-24
CN106153915A (en) 2016-11-23

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