CN111363851A - 一种检测新型冠状病毒的方法和试剂盒 - Google Patents
一种检测新型冠状病毒的方法和试剂盒 Download PDFInfo
- Publication number
- CN111363851A CN111363851A CN202010301619.1A CN202010301619A CN111363851A CN 111363851 A CN111363851 A CN 111363851A CN 202010301619 A CN202010301619 A CN 202010301619A CN 111363851 A CN111363851 A CN 111363851A
- Authority
- CN
- China
- Prior art keywords
- probe
- novel coronavirus
- primer
- pseudovirus
- kit
- 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
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 54
- 238000000034 method Methods 0.000 title claims abstract description 13
- 239000000523 sample Substances 0.000 claims abstract description 47
- 238000007847 digital PCR Methods 0.000 claims abstract description 24
- 238000006243 chemical reaction Methods 0.000 claims abstract description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 18
- 108010085238 Actins Proteins 0.000 claims description 16
- 102100034343 Integrase Human genes 0.000 claims description 11
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 claims description 11
- 238000011144 upstream manufacturing Methods 0.000 claims description 10
- 102000007469 Actins Human genes 0.000 claims description 6
- 238000010791 quenching Methods 0.000 claims description 6
- 230000000171 quenching effect Effects 0.000 claims description 4
- 125000006853 reporter group Chemical group 0.000 claims description 4
- 238000003752 polymerase chain reaction Methods 0.000 claims description 3
- UDGUGZTYGWUUSG-UHFFFAOYSA-N 4-[4-[[2,5-dimethoxy-4-[(4-nitrophenyl)diazenyl]phenyl]diazenyl]-n-methylanilino]butanoic acid Chemical compound COC=1C=C(N=NC=2C=CC(=CC=2)N(C)CCCC(O)=O)C(OC)=CC=1N=NC1=CC=C([N+]([O-])=O)C=C1 UDGUGZTYGWUUSG-UHFFFAOYSA-N 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 26
- 108020004707 nucleic acids Proteins 0.000 abstract description 7
- 150000007523 nucleic acids Chemical class 0.000 abstract description 7
- 102000039446 nucleic acids Human genes 0.000 abstract description 7
- 244000052769 pathogen Species 0.000 abstract description 7
- 230000001717 pathogenic effect Effects 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 238000011002 quantification Methods 0.000 abstract description 4
- 241001112090 Pseudovirus Species 0.000 description 67
- 239000013558 reference substance Substances 0.000 description 25
- 238000002156 mixing Methods 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 6
- 238000003786 synthesis reaction Methods 0.000 description 6
- 230000035772 mutation Effects 0.000 description 5
- 241000700605 Viruses Species 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000013461 design Methods 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 206010060891 General symptom Diseases 0.000 description 1
- HEFNNWSXXWATRW-UHFFFAOYSA-N Ibuprofen Chemical compound CC(C)CC1=CC=C(C(C)C(O)=O)C=C1 HEFNNWSXXWATRW-UHFFFAOYSA-N 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 1
- 208000004756 Respiratory Insufficiency Diseases 0.000 description 1
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 208000017574 dry cough Diseases 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000013074 reference sample Substances 0.000 description 1
- 201000004193 respiratory failure Diseases 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 208000026425 severe pneumonia Diseases 0.000 description 1
- 238000002636 symptomatic treatment Methods 0.000 description 1
Images
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
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
-
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Description
QuantStudioTM 3D Digital PCR Master Mix v2(2×) | 7.3μL |
新型冠状病毒引物探针混合液(20×) | 0.7μL |
HiScript III Reverse Transcriptase(200U/μL) | 2μL |
假病毒模板 | 4.5μL |
样本名称 | 新型冠状假病毒含量检测结果 |
假病毒含量参考品-100000copies/ml | 131000.43copies/ml |
假病毒含量参考品-10000copies/ml | 8652.28copies/ml |
假病毒含量参考品-1000copies/ml | 1280.75copies/ml |
假病毒含量参考品-100copies/ml | 170.71copies/ml |
假病毒含量参考品-50copies/ml | 70.18copies/ml |
样本名称 | 新型冠状假病毒含量检测结果 |
假病毒含量参考品-50copies/ml | 70.18copies/ml |
假病毒含量参考品-50copies/ml | 40.98copies/ml |
假病毒含量参考品-50copies/ml | 70.55copies/ml |
假病毒含量参考品-50copies/ml | 60.05copies/ml |
假病毒含量参考品-50copies/ml | 41.01copies/ml |
假病毒含量参考品-50copies/ml | 59.22copies/ml |
假病毒含量参考品-50copies/ml | 62.17copies/ml |
假病毒含量参考品-50copies/ml | 53.97copies/ml |
假病毒含量参考品-50copies/ml | 68.55copies/ml |
假病毒含量参考品-50copies/ml | 56.72copies/ml |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 10.88copies/ml |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 35.22copies/ml |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 未检出 |
假病毒含量参考品-25copies/ml | 未检出 |
Claims (9)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010301619.1A CN111363851A (zh) | 2020-04-16 | 2020-04-16 | 一种检测新型冠状病毒的方法和试剂盒 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010301619.1A CN111363851A (zh) | 2020-04-16 | 2020-04-16 | 一种检测新型冠状病毒的方法和试剂盒 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN111363851A true CN111363851A (zh) | 2020-07-03 |
Family
ID=71203298
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010301619.1A Pending CN111363851A (zh) | 2020-04-16 | 2020-04-16 | 一种检测新型冠状病毒的方法和试剂盒 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN111363851A (zh) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113049818A (zh) * | 2021-01-11 | 2021-06-29 | 广东菲鹏生物有限公司 | 一种鉴别突变型抗原的方法及试剂 |
CN113073150A (zh) * | 2021-04-28 | 2021-07-06 | 领航基因科技(杭州)有限公司 | 一种用于新型冠状病毒及其变异株的数字pcr检测试剂盒 |
CN113215231A (zh) * | 2021-03-30 | 2021-08-06 | 成都里来生物科技有限公司 | 一种SARS-CoV和COVID-19病毒双重PCR检测方法 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107190091A (zh) * | 2017-07-21 | 2017-09-22 | 大连晶泰生物技术有限公司 | 适用于ffpe 样品的基因表达定量的实时定量pcr 方法 |
CN110982945A (zh) * | 2020-03-04 | 2020-04-10 | 珠海丽珠试剂股份有限公司 | 一种检测2019新型冠状病毒的核酸组合物、试剂盒以及方法 |
CN110982943A (zh) * | 2020-03-03 | 2020-04-10 | 中山大学达安基因股份有限公司 | 一种新型冠状病毒rt-pcr检测方法及试剂盒 |
-
2020
- 2020-04-16 CN CN202010301619.1A patent/CN111363851A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107190091A (zh) * | 2017-07-21 | 2017-09-22 | 大连晶泰生物技术有限公司 | 适用于ffpe 样品的基因表达定量的实时定量pcr 方法 |
CN110982943A (zh) * | 2020-03-03 | 2020-04-10 | 中山大学达安基因股份有限公司 | 一种新型冠状病毒rt-pcr检测方法及试剂盒 |
CN110982945A (zh) * | 2020-03-04 | 2020-04-10 | 珠海丽珠试剂股份有限公司 | 一种检测2019新型冠状病毒的核酸组合物、试剂盒以及方法 |
Non-Patent Citations (1)
Title |
---|
FENGTING YU ET AL.: "Quantitative Detection and Viral Load Analysis of SARS-CoV-2 in Infected Patients", 《CLINICAL INFECTIOUS DISEASES》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113049818A (zh) * | 2021-01-11 | 2021-06-29 | 广东菲鹏生物有限公司 | 一种鉴别突变型抗原的方法及试剂 |
CN113215231A (zh) * | 2021-03-30 | 2021-08-06 | 成都里来生物科技有限公司 | 一种SARS-CoV和COVID-19病毒双重PCR检测方法 |
CN113215231B (zh) * | 2021-03-30 | 2022-08-05 | 成都里来生物科技有限公司 | 一种SARS-CoV和COVID-19病毒双重PCR检测方法 |
CN113073150A (zh) * | 2021-04-28 | 2021-07-06 | 领航基因科技(杭州)有限公司 | 一种用于新型冠状病毒及其变异株的数字pcr检测试剂盒 |
CN113073150B (zh) * | 2021-04-28 | 2023-01-10 | 领航医学科技(深圳)有限公司 | 一种用于新型冠状病毒及其变异株的数字pcr检测试剂盒 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN111041129B (zh) | 检测6项呼吸道病毒的引物探针组合、试剂盒及应用 | |
Mollaei et al. | Comparison five primer sets from different genome region of COVID-19 for detection of virus infection by conventional RT-PCR | |
CN111363851A (zh) | 一种检测新型冠状病毒的方法和试剂盒 | |
CN112063756B (zh) | 多重检测呼吸道病毒核酸的方法及试剂盒 | |
CN113215312A (zh) | 一种冠状病毒SARS-CoV-2数字PCR多重检测试剂盒及其应用 | |
US8119338B2 (en) | Methods and compositions for detecting rhinoviruses | |
CN106319055B (zh) | 一种三联核酸检测试剂盒 | |
CN111286559B (zh) | 检测非洲猪瘟病毒的引物、探针及试剂盒 | |
CN111560478B (zh) | 一种一步法反转录PCR结合Sanger测序检测新型冠状病毒的试剂盒 | |
CN105886663B (zh) | 一种锁核酸增敏的猪伪狂犬病毒野毒株荧光定量pcr的检测试剂盒 | |
CN112813195B (zh) | 基于微液滴数字分析的新型冠状病毒核酸定量检测试剂盒 | |
CN110305987A (zh) | 检测诺如病毒i型、诺如病毒ii型和轮状病毒的试剂盒 | |
CN112760421A (zh) | 同时检测牛轮状病毒、冠状病毒和病毒性腹泻病毒的三重荧光定量pcr试剂盒及应用方法 | |
CN112646932B (zh) | 一步法可视化检测新型冠状病毒核酸的引物组和试剂盒 | |
CN114457197A (zh) | Raa荧光法检测fpv的引物探针组及试剂盒 | |
WO2024055627A1 (zh) | 一种用于猴痘病毒野生型和突变型分子分型的引物分子信标组合及其应用 | |
CN117757985A (zh) | 用于冠状病毒检测的组合物及其制备和使用方法 | |
CN116814859A (zh) | 鉴别非洲猪瘟病毒基因ⅰ型和ⅱ型的引物探针组合物、试剂盒及方法 | |
CN111172301A (zh) | 一种艰难梭菌毒素b的pcr荧光检测试剂盒及其应用 | |
CN111471800A (zh) | 检测新型冠状病毒的试剂盒及其扩增引物组合物 | |
CN115927746A (zh) | 用于诺如病毒和轮状病毒双重rt-rap检测的引物探针组、试剂盒及其应用 | |
AU2021103861A4 (en) | Method and kit for differentially detecting porcine pseudorabies vaccine virus and wild virus | |
AU2020104066A4 (en) | Double fluorescent quantitative RT-PCR detection method for Classical swine fever virus and Bovine viral diarrhea virus | |
CN106521030A (zh) | 猪瘟病毒、牛病毒性腹泻病毒双重荧光定量rt‑pcr检测方法 | |
CN115838837B (zh) | 一种检测猪德尔塔冠状病毒和猪流行性腹泻病毒的微滴式双重数字pcr试剂盒 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
TA01 | Transfer of patent application right |
Effective date of registration: 20220507 Address after: 116600 no.9-2 Jinqi Road, Dalian Economic and Technological Development Zone, Liaoning Province Applicant after: Dalian Jingtai medical laboratory Co.,Ltd. Applicant after: JIANGSU MACRO&MICRO-TEST MED-TECH Co.,Ltd. Address before: 116635 No. 9, Jinqi Road, advanced equipment manufacturing park, Jinzhou new area, Dalian City, Liaoning Province Applicant before: DALIAN GENTALKER BIOTECHNOLOGY Co.,Ltd. Applicant before: JIANGSU MACRO&MICRO-TEST MED-TECH Co.,Ltd. |
|
TA01 | Transfer of patent application right | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200703 |
|
RJ01 | Rejection of invention patent application after publication |