CN105891480A - Stomach disease detection kit - Google Patents

Stomach disease detection kit Download PDF

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Publication number
CN105891480A
CN105891480A CN201610504850.4A CN201610504850A CN105891480A CN 105891480 A CN105891480 A CN 105891480A CN 201610504850 A CN201610504850 A CN 201610504850A CN 105891480 A CN105891480 A CN 105891480A
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China
Prior art keywords
detection kit
disease detection
nucleic acid
stomach disease
pepsinogen
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CN201610504850.4A
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Chinese (zh)
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卢美珍
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Individual
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Priority to CN201610504850.4A priority Critical patent/CN105891480A/en
Publication of CN105891480A publication Critical patent/CN105891480A/en
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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

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 November 29, Application No. 201510849189.6, invention name in 2015 It is referred to as the divisional application of the application for a patent for invention of " a kind of test kit for detection of stomach disorders ".
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, The PGII of gastric mucosa synthesis is about the 25% of total amount, and PG II major part enters digestive tract, and a small amount of blood that enters follows Ring, 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 it raises and at the bottom of stomach Glandular tube atrophy, intestinal epithelial metaplasia or Pseudopyloric gland metaplasia, dysplasia are relevant.Measure PG II content to help Detect other digestive tract disease such as duodenal ulcer, atrophic gastritis, gastritis, gastric cancer, examine for clinic Survey and health check-up examination demand.In crowd's gastropathy examination, everyone makees gastroscope is unpractical, can be invaded by non- Entering property serum PG detects superficial gastritis, erosive gastritis, gastric ulcer, duodenal ulcer, gastric cancer etc. Out, then to carry out gastroscopy be a kind of realistic plan to high-risk group's examination.Employing known in the art Time-resolved fluorescence measures technology, and the TRFIIA test kit of the PGII of establishment is in current PGII detection 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 the one grown up early 1990s New combinatorial chemistry technique, it uses jumbo random oligonucleotide library, the outer PCR amplification technique of coalition, With the oligonucleotide of index concentration Yu target molecule specific bond, through multi-turns screen, it is thus achieved that affinity is high, special The oligonucleotide aptamer (aptamers) that property is strong.The own Successful utilization of this technology in the screening of many target molecules, Including metal ion, organic dyestuff, medicine, protein, aminoacid and various cytokines etc..
Summary of the invention
It is an object of the invention to screen pepsinogen by phyletic evolution index concentration technology (SELEX technology) The oligonucleotide aptamer of II substitutes antibody, it is provided that a kind of succinct quick, high sensitivity, high specific Pepsinogen I I detection and isolation and purification method in early days.
Technical scheme:
For single stranded DNA random library and the primer of phyletic evolution index concentration technology screening in the present invention, by U.S. Invitrogen company of state 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 according to the recombined human pepsinogen I I isozyme disclosed in CN103387971A The method of chimeric protein 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 DNA molecular shown in NO:1-17 is arbitrary.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: there is identical function Oligonucleotide 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 There is identical function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: for energy and DNA sequence Carry out the oligonucleotide sequence hybridized.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: at nucleic acid aptamer Oligonucleotide aptamer sequence obtained by the deletion of any position or increase part oligonucleotide residues has identical Function.
A kind of above-mentioned affine human pepsinogen II nucleic acid aptamer, it is characterised in that: at nucleic acid aptamer After any position carries 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: at nucleic acid aptamer Any position carry out phosphorylation, methylate, amino, sulfydryl, isotope, biotin, digoxin, fluorescence After matter, nano luminescent material or enzyme labelling are 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 people's pepsin The detection of former II and isolated and purified, thus 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) synthesis of single stranded DNA random oligonucleotide library；2) utilize phyletic evolution index concentration method to oligonucleoside Acid library is screened；3) amplification and the oligonucleotide of human albumin specific bond；4) next round screening is carried out, Purpose oligonucleotide sequence is obtained after 12 take turns above screening；5) cloning and sequencing.
The invention have the advantage that there is simplicity, quick, economic dispatch feature, with other combinatorial chemical libraries as random Peptide storehouse, antibody library are compared with phage display libraries, the aptamer filtered out from oligonucleotide library The many advantages of tool: 1) itself it is oligonucleotide, molecular weight, can be cost-effective with chemosynthesis；2) There is affinity more higher than antibody and specificity；3) it is easy to labelling and selectively can mark at different parts Note；4) repeatability and good stability, and be prone to preserve, i.e. insensitive to high temperature and drastic conditions.Therefore, it is System evolution index concentration technology has a good application prospect.
Detailed description of the invention
The preparation of embodiment 1 human pepsinogen II
Method according to the recombined human pepsinogen I I isozyme chimeric protein disclosed in CN103387971A Preparing 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 a length of 82nt SsDNA pool, two ends are immobilized primer sequence, and centre is the random sequence of 42 bases, storage capacity Amount is I014Above；Primer 1:5'TACGACATGAACCGTGATAA-3'；Primer 2: 5'- TCGATCAGCAGGTTTCACTG-3'；SsDNA pool and two kinds of primers are all become with TE buffer 100 μm ol/L stock solution _ 20 DEG C storages are standby.
Being double-stranded DNA by single-stranded DNA banks amplification, product is through 2% agarose gel electrophoresis and cuts glue recovery purification； With reclaim double-stranded DNA as template, in vitro transcription goes out single stranded RNA random library, and transcription product is pure through PAGE Change.The RNA molecule being combined with film is removed, then with 2ug through anti-sieve of nitrocellulose filter in 75 μ g RNA libraries Human pepsinogen II albumen, hatches 30min for 37 DEG C, and reactant liquor filters through nitrocellulose filter, washing filter Film；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, external turn Record out RNA library to screen for next round；Often in wheel screening process, RT-PCR obtains double-stranded DNA library, with This double-stranded DNA is that template in vitro transcription goes out RNA aptamer storehouse, and screening carries out 10 altogether and takes turns.Obtain 14 to fit Gamete, its sequence is respectively shown in SEQ ID NO:1-14.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, digests lh, purification with calf intestinal alkaline phosphatase (CIP) 37 DEG C Reclaim dephosphorylized RNA；By T4 polynucleotide kinase labelling [γ-32P] ATP in dephosphorylized RNA Molecular end.The radiolabeled aptamer of 10nmol respectively with people's pepsin of variable concentrations (1-200nM) Proenzyme II 37 DEG C hatches 30min, and each group reactant liquor filters through nitrocellulose filter, washs filter membrane, is dried filter Film, liquid scintillation counter measures the exit dose of residual on filter membrane, and same sample is parallel does twice mensuration.Calculate each Aptamer and the dissociation constant of 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, Pepsinogen I I, carries out specific detection with 14 aptamers, finds through binding tests, and these are adaptive Son does not combines with these albumen, and only keeps 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. Detected by RT-PCR, find its Stability Analysis of Structures of placement of surrounding, be 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, sample with 9 patients and 4 normal persons mixes respectively 40min, is separated by biotin, the content of quantitative analysis human pepsinogen therein II, sends out by analyzing Existing, in 9 patients, the content of pepsinogen I I dramatically increases, and has exceeded the threshold value of regulation.Reach phase Answer the diagnostic criteria that 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 the skill of this area For art personnel, all any modification, equivalent substitution and improvement etc. done within the spirit and principles in the present invention, Should be included within the scope of the present 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:13.
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.
CN201610504850.4A 2015-11-29 2015-11-29 Stomach disease detection kit Pending CN105891480A (en)

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CN201510849189.6A CN105277698B (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504850.4A CN105891480A (en) 2015-11-29 2015-11-29 Stomach disease detection kit

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CN201610497173.8A Withdrawn CN106153914A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504404.3A Withdrawn CN106405083A (en) 2015-11-29 2015-11-29 A 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
CN201610505390.7A Pending CN105929161A (en) 2015-11-29 2015-11-29 Kit for detecting stomach diseases
CN201610504405.8A Pending CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610503246.XA Pending CN106053809A (en) 2015-11-29 2015-11-29 Kit for detecting gastric disease
CN201610504850.4A Pending CN105891480A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610505427.6A Withdrawn CN106153916A (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
CN201610503443.1A Withdrawn CN106198975A (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
CN201610504437.8A Pending CN105929160A (en) 2015-11-29 2015-11-29 Kit for stomach disease detection
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

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CN201610497173.8A Withdrawn CN106153914A (en) 2015-11-29 2015-11-29 A kind of test kit for detection of stomach disorders
CN201610504404.3A Withdrawn CN106405083A (en) 2015-11-29 2015-11-29 A 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
CN201610505390.7A Pending CN105929161A (en) 2015-11-29 2015-11-29 Kit for detecting stomach diseases
CN201610504405.8A Pending CN105891479A (en) 2015-11-29 2015-11-29 Stomach disease detection kit
CN201610503246.XA Pending CN106053809A (en) 2015-11-29 2015-11-29 Kit for detecting gastric disease

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CN201610505427.6A Withdrawn CN106153916A (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
CN201610503443.1A Withdrawn CN106198975A (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
CN201610504437.8A Pending CN105929160A (en) 2015-11-29 2015-11-29 Kit for stomach disease detection
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

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