CN102108391A - Detection kit for drug resistance mutation of adefovir dipivoxil - Google Patents
Detection kit for drug resistance mutation of adefovir dipivoxil Download PDFInfo
- Publication number
- CN102108391A CN102108391A CN2009102004966A CN200910200496A CN102108391A CN 102108391 A CN102108391 A CN 102108391A CN 2009102004966 A CN2009102004966 A CN 2009102004966A CN 200910200496 A CN200910200496 A CN 200910200496A CN 102108391 A CN102108391 A CN 102108391A
- Authority
- CN
- China
- Prior art keywords
- drug resistance
- adefovir dipivoxil
- detecting
- gene
- resistance mutation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kit for detecting the drug resistance genetic mutation of individual adefovir dipivoxil. The kit comprises a specific primer pair and a specific fluorescent probe pair for detecting the polymorphism of the P gene rtN236T site of the adefovir dipivoxil drug resistance mutation site, conventional components of fluorescent quantitation PCR (Polymerase Chain Reaction), and the like. The drug resistance of the individual adefovir dipivoxil is evaluated by detecting the polymorphism of the P gene rtN236T site by the kit disclosed by the invention.
Description
Technical field
The present invention relates to molecular biology and medical field, more specifically, the present invention relates to a kind of test kit that detects individual adefovir ester resistance mutation genetic, assess individual adefovir ester thing resistance by detecting adefovir ester medication target site gene pleiomorphism.
Background technology
Adefovir ester is the oral precursor of the lipotropy of Adefovir, and its intestinal absorption rate and bioavailability are all higher, and oral back is converted into Adefovir rapidly under the nonspecific esterase hydrolytic action.Adefovir is a kind of acyclic dAMP analogue, further is converted into activated bis phosphoric acid Adefovir in vivo and an antivirus action.Experimental study and clinical trial prove that all adefovir ester can reduce the concentration of hepatitis B virus DNA rapidly, alleviates the pathology of liver organization.
The life-time service adefovir ester can be induced the hepatitis B virus variation, and produces resistance, and the hepatitis B virus DNA of having turned out cloudy in the serum is reappeared, even with disease relapse.
Present known adefovir ester resistance major cause is the variation of rtN236T site.The rtN236T variation then is to be caused to the C point mutation by P gene the 836th bit base A, and the variation of hydrogen bond between base has promptly taken place.
Summary of the invention
The invention provides a kind of test kit that detects individual adefovir ester resistance mutation genetic.
This test kit comprises: the Auele Specific Primer that detects adefovir ester resistance variant sites P gene rtN236T loci polymorphism is right to reaching the specificity fluorescent probe, Taq enzyme, dNTP mixed solution, MgCl
2Solution, reaction buffer, deionized water.
Auele Specific Primer described in this test kit and fluorescent probe can be finished design for the person of ordinary skill of the art easily, and Auele Specific Primer and fluorescent probe synthesize the DNA synthetic technology of available routine.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.The experimental technique of unreceipted actual conditions in the following example, usually according to normal condition, or the condition of advising according to manufacturer.
The use of embodiment detection kit
The extraction of step 1:DNA template
Genomic dna with silica gel adsorption extracting mouth epithelial cells.
Step 2: quantitative fluorescent PCR reaction
Use the quantitative fluorescent PCR suit in the detection kit, carry out the quantitative fluorescent PCR reaction, the system of reaction is cumulative volume 10 μ l, and comprising concentration is dna profiling 2 μ l, 1 μ l 10X quantitative fluorescent PCR reaction buffer, 0.1 μ l 25mM dNTP mixed solution, the 0.6 μ l 25mMMgCl of 20ng/ μ l
2The band VIC fluorescent probe that adopted primer and antisense primer each 0.225 μ l, 10 μ M are arranged of solution, 0.025 μ l (5units/ μ l) Taq archaeal dna polymerase, 20 μ M and each 0.25 μ l of band FAM fluorescent probe, deionized water 5.325 μ l.
React on the pcr amplification instrument, reaction conditions is 50 ℃, 2 minutes, 95 ℃, 10 minutes, carries out 95 ℃ of 60 round-robin, 30 seconds, 60 ℃, 1 minute.Reaction finishes the back and read the fluorescent amount on quantitative real time PCR Instrument.
Step 4: gene type assay
According to the gene type diagram that the test kit working instructions indicate gene type assay is carried out in the SNP site.
The those skilled in the art that are familiar with fluorescent quantitative PCR technique can be by the final sample fluorescence volume that shows on the identification quantitative real time PCR Instrument, can determine the genotype in the SNP site detected according to the power of different sequence fluorescence probe signals.
Claims (1)
1. a test kit that detects individual adefovir ester resistance mutation genetic comprises that Auele Specific Primer is right to reaching the specificity fluorescent probe, Taq enzyme, dNTP mixed solution, MgCl
2Solution, quantitative fluorescent PCR reaction buffer and deionized water is characterized in that: described Auele Specific Primer is to reaching the specificity fluorescent probe to designing at detecting P gene rtN236T loci polymorphism.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102004966A CN102108391A (en) | 2009-12-23 | 2009-12-23 | Detection kit for drug resistance mutation of adefovir dipivoxil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009102004966A CN102108391A (en) | 2009-12-23 | 2009-12-23 | Detection kit for drug resistance mutation of adefovir dipivoxil |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102108391A true CN102108391A (en) | 2011-06-29 |
Family
ID=44172676
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009102004966A Pending CN102108391A (en) | 2009-12-23 | 2009-12-23 | Detection kit for drug resistance mutation of adefovir dipivoxil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102108391A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1723907A (en) * | 2005-07-01 | 2006-01-25 | 肖春 | Medicinal composition for treating chronic hepatitis B |
| CN101004415A (en) * | 2007-01-16 | 2007-07-25 | 复旦大学附属华山医院 | Method for measuring drug fast variation of hepatitis b virus to Adefovir |
-
2009
- 2009-12-23 CN CN2009102004966A patent/CN102108391A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1723907A (en) * | 2005-07-01 | 2006-01-25 | 肖春 | Medicinal composition for treating chronic hepatitis B |
| CN101004415A (en) * | 2007-01-16 | 2007-07-25 | 复旦大学附属华山医院 | Method for measuring drug fast variation of hepatitis b virus to Adefovir |
Non-Patent Citations (1)
| Title |
|---|
| 王磊等: "应用巢式聚合酶链反应－限制性片段长度多态性技术检测乙型肝炎病毒阿德福韦耐药变异rtN236T", 《胃肠病学和肝病学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4945779B2 (en) | Method | |
| JP2020202831A (en) | Methods for Detecting Nucleic Acid Sequence Variants | |
| EP1716257B8 (en) | New primers and probes for the detection of parvovirus b19 | |
| CN102433376B (en) | Fluorescence quenching-based genetic variation detection method and probe | |
| KR20090058451A (en) | Method for selective labeling and detection of target nucleic acids using immobilized peptide nucleic acid probes | |
| US9353404B2 (en) | Capture based nucleic acid detection | |
| RU2018122965A (en) | IMPROVED METHOD AND KIT FOR DETERMINING SEVERITY AND PROGRESSING PERIODONTAL DISEASE | |
| CN102229989B (en) | Method and kit for detecting ethambutol resistance mutation of Mycobacterium tuberculosis | |
| WO2009061640A3 (en) | Hepatitis b virus (hbv) specific oligonucleotide sequences | |
| CN103045717B (en) | Pyrosequencing techniques detects the method for mycobacterium tuberculosis detection of rifampin resistant | |
| CN101608219A (en) | Hypertension genetic test kit | |
| CN102586409A (en) | Multiple reverse transcription polymerase chain reaction detection method for swine transmissible gastroenteritis | |
| CN103045716B (en) | Method for detecting isoniazid drug resistance of mycobacterium tuberculosis by using pyrosequencing technology | |
| JP6164683B2 (en) | Method for analyzing deletion site of spermatogenesis region on Y chromosome and Y chromosome | |
| WO2004050911A3 (en) | Method for lowering both sequence variations and increase of base lines effects in diagnostic hybridisation assay, assay for performing such a method and probe for use in the assay | |
| Zhang et al. | RART-LAMP: One-Step Extraction-Free Method for Genotyping within 40 min | |
| CN102108391A (en) | Detection kit for drug resistance mutation of adefovir dipivoxil | |
| CA2549059A1 (en) | Oligonucleotides for the detection of hepatitis b virus | |
| ES2445709T3 (en) | Method for the identification by molecular techniques of genetic variants that do not encode D (D-) antigen and encode altered C (C + W) antigen | |
| CN103667447A (en) | Primer and kit for detecting NQO1 gene polymorphism, and application of primer and kit on pathological examination | |
| Taskova et al. | Tandem oligonucleotide probe annealing and elongation to discriminate viral sequence | |
| CN102108390A (en) | Entecavir resistant mutation test kit | |
| CN101608229A (en) | Kit for genetic detection of gout | |
| CN102108392A (en) | Detection for drug-resistant mutation heredity of hepatitis B virus nucleoside analogue | |
| WO2012048339A3 (en) | Selective detection of neisseria meningitidis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C53 | Correction of patent of invention or patent application | ||
| CB02 | Change of applicant information |
Address after: Ginza wealth International Plaza No. 43 Shanghai Guoquan road 200433 4 floor Applicant after: XINBAXIANG (SHANGHAI) MOLECULAR MEDICAL TECHNOLOGY (SHANGHAI) CO., LTD. Address before: 200433 No. 335, building 2, No. 6, National Road, Shanghai Applicant before: Shanghai Zhujian Bioengineering Co., Ltd. |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: SHANGHAI ZHUJIAN BIOENGINEERING CO., LTD. TO: INSTITUTE OF TARGETED THERAPY AND MOLECULAR MEDICINE OF SHANGHAI |
|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20110629 |