CN110317864A - A method of it is sequenced by macro transcript profile to detect pathogen - Google Patents
A method of it is sequenced by macro transcript profile to detect pathogen Download PDFInfo
- Publication number
- CN110317864A CN110317864A CN201910652791.9A CN201910652791A CN110317864A CN 110317864 A CN110317864 A CN 110317864A CN 201910652791 A CN201910652791 A CN 201910652791A CN 110317864 A CN110317864 A CN 110317864A
- Authority
- CN
- China
- Prior art keywords
- rna
- pathogen
- data
- host
- rrna
- 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
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
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provide it is a kind of by macro transcript profile be sequenced detection pathogen method, the method by only detect RNA mode come and meanwhile detect DNA and RNA pathogen.The problems such as data volume after this set process compared to conventional cause of disease NGS test can decline by the method for the invention, solve existing DNA, RNA and build library detection respectively, and market expense is high, and data are many and diverse, analysis difficulty height and clinical missing inspection.
Description
Technical field
The present invention relates to pathogen detection fields, and in particular to a kind of to be sequenced by macro transcript profile to detect the side of pathogen
Method.
Background technique
Cause of disease NGS testing product on the market at present, basic only detection DNA pathogen, does not detect RNA pathogen, clinical
Using there are a large amount of missing inspections；It can be directed to DNA Pathogen test DNA, RNA Pathogen test RNA in scientific research, it is same separately to construct 2 libraries
When detect, complex steps, output data amount is many and diverse, and expense is high, and data are analyzed difficulty and increased, and calculation resources consumption is big, market
It is difficult to receive.
The study found that the Total-RNA that we extract in Pathogen test wherein host's rRNA (including mitochondria
RRNA 16s, 12s rRNA；Cytoplasm rRNA28s, 5.8s, 18s, 5srRNA) account for the 80-90% of total serum IgE ratio, remaining master
Wanting composition is (mRNAglobin mRNA, mammal height express conservative gene GAPDH etc.) of host；The RNA of pathogen,
The trace dnas such as tRNA, lncRNA, miRNA only occupy about 1%.If we can get rid of rRNA and part is high
The conservative mRNA of expression, in this way has the Sensitivity of our pathogen high promotion.
In addition, can all be transcribed substantially according to central dogma life entity, we have found in many experiments, same 1ml human blood
DNA nucleic acid is extracted probably in 30 micrograms, extracts RNA nucleic acid probably in 3 micrograms, transcript accounts for the 1-2% of genome,
The copy number of the conversion general RNA and DNA of copy number multiple proportion is horizontal in the same order of magnitude.We utilize fluorescent quantitation simultaneously
It includes hepatitis B, human papilloma virus, herpes simplex virus I-type, cloth Lu Shi bar that round pcr, which tests clinical infection patient,
Bacterium, white to read coccus, comprising virus, bacterium, a variety of sample types of fungi detect its DNA and RNA respectively, and discovery RNA detects CT value
CT value is detected far below DNA, for difference in 4-10 etc., the copy number discovery that converts detects the transcript folder RNA ratio DNA spirit of the cause of disease
Sensitivity is 10-1000 times high, depending on Different Kinds of Pathogens.
Summary of the invention
According to above-mentioned discovery, after eliminating host RNA in method of the invention, by only detecting RNA after removal host RNA
Mode, pathogen RNA account for total-RNA ratio improve；DNA pathogen is detected by transcript folder RNA and can be improved sensitive
Degree, the method for the present invention is a kind of sensitive testing process of height for only surveying RNA to detect all pathogen, by comparing after this set process
It can decline in the data volume of conventional cause of disease NGS test, solve existing DNA, RNA and build library detection respectively, market expense is high,
The problems such as data are many and diverse, analysis difficulty height and clinical missing inspection, and have higher detection sensitivity for infection type sample.(turn
Record originally can reflect the active degree of the pathogenic bacteria in sample)
It is to grind that macro transcript profile (Metatranscriptome) sequencing, which is with whole RNA of microbiologic population in specific sample,
Study carefully object, enlivens the composition of strain and the expression of active gene from analyzing in microbiologic population on transcriptional level.Macro turn
Record group can also study the composition situation for enlivening strain and cance high-expression gene, disclose specific ring while carrying out species identification
The adaptability of strain and the possible regulatory mechanism of gene expression under the Effects of Factors of border.In addition, macro transcript profile sequencing research is avoided
The process that microorganism is separately cultured, extend microbial resources utilizes space, studies and has provided for the transcription of microorganism
Effect tool.
In one embodiment, the present invention provides a kind of method that detection pathogen is sequenced by macro transcript profile, described
Method by only detect RNA mode come and meanwhile detect DNA and RNA pathogen.
In one embodiment, the described method comprises the following steps: step 1 is prepared from biological sample to be measured
Total-RNA sample；Step 2 removes host RNA using host's probe cell of design, constructs RNA high-throughput sequencing library；Step
Rapid three, high-flux sequence is carried out using the RNA high-throughput sequencing library, obtains RNA sequencing data；And step 4, to described
RNA sequencing data carries out bioinformatic analysis, determines whether the biological sample to be measured contains pathogen to be detected.
In one embodiment, the biological sample to be measured is selected from peripheral blood, lesion tissue, cerebrospinal fluid, alveolar washing
Liquid, Pleural effusions, sputum, respiratory secretions or digestive tract secretion.
In one embodiment, the high-flux sequence is surveyed using 500 sequenator of illumina Nextseq
Sequence.
In one embodiment, it includes conservative rRNA, globulin mRNA that host RNA is removed in the step 2
MRNA is expressed with host's height.
In one embodiment, the conservative rRNA includes: mitochondria rRNA 16s rRNA, 12s rRNA；
Cytoplasm rRNA 28s rRNA, 5.8s rRNA, 18s rRNA, 5s rRNA.
In one embodiment, the step 4 the following steps are included:
A. Adapter, Duplication, Poly structure are removed from the RNA sequencing data and filter quality value is lower than
The reads of Q20, removal length are less than the reads of 50bp, obtain valid data；
B. the valid data are compared with host genome data, the reads compared therewith are removed,
Host data amount is counted, nonhost data are retained, obtains and compares to the reads quantity N on hosthWith host genome length Lh；
C. the nonhost data are compared with the property data base of pathogen to be detected, obtain represent compare to
Reads quantity, pathogen N to be detected on detection pathogen iiGenome length Li, screen comparison result, determine it is described to
Survey whether biological sample contains pathogen to be detected.
In one embodiment, if the biological sample contain pathogen to be detected, by the cause of disease data use with
Lower calculation formula is standardized, and determines pathogen detection value Ti,
In one embodiment, the host genome data are full-length genome data；The valid data and host
Genomic data is compared by bowtie bwa software.
In one embodiment, the pathogen to be detected is helminth, fungi, bacterium, actinomyces, virus, Zhi Yuan
Body, Chlamydia, Richettsia or conveyor screw.
High-throughput RNA sequencing library of the invention carries out high-flux sequence and refers to the data volume need according to pathogen detection
It asks, treats the sequencing library Index and carry out upper machine sequencing after certain proportion mixing, it is final to obtain high-flux sequence data.The present invention
Method the purpose that may be implemented to detect DNA, RNA pathogen simultaneously is only realized with the detection of a set of process, measured in various high passes
The process can be used on sequence instrument, cost can be effectively reduced, while improving detection sensitivity, it is comprehensive.
Method of the invention is a kind of sensitive testing process of height for only surveying RNA and coming while detecting all pathogen, passes through this
Data volume after set process compared to conventional cause of disease NGS test can decline, and solve existing DNA, RNA and build library detection respectively,
The problems such as market expense is high, and data are many and diverse, analysis difficulty height and clinical missing inspection, and have higher inspection for infection type sample
Survey sensitivity.(active degree that transcript can reflect the pathogenic bacteria in sample)
Specific embodiment
In order to make those skilled in the art more fully understand the technical solution in the application, below in conjunction with embodiment to this
Invention is described further, it is clear that described embodiments are only a part of embodiments of the present application, rather than whole implementation
Example.Based on the embodiment in the application, obtained by those of ordinary skill in the art without making creative efforts
All other embodiment, shall fall within the protection scope of the present application.It is routine unless otherwise specified in following embodiment
Method.
In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition, such as Joseph
Written by Sambrook et al., condition described in " Molecular Cloning:A Laboratory guide (third edition) ", or by proposed by manufacturer
Condition.Used various chemical reagent, are commercial product in embodiment.
Embodiment one: the method for the present invention and DNA library method compare
The analog sample for testing known infection type, extracts be mixed with DNA virus herpes simplex virus type 1 (HSV1) respectively
DNA, RNA of blood simulated infection sample and the blood simulated infection sample for being mixed with bacterium (Escherichia coli), construct standard respectively
DNA library and the library the method for the present invention RNA, and it is mixed with the blood simulated infection of RNA virus Hepatitis C Virus (HCV)
Sample and the blood simulated infection sample rna for being mixed with RNA virus japanese encephalitis virus (JEV), building the method for the present invention RNA text
Library, the Pathogen test situation of 2 kinds of methods of contrast verification.
A: the method for the present invention testing process:
1. extracting above-mentioned 4 parts of sample total-RNA with RNeasy Mini Kit (Qiagen)；
2. host's probe cell using design removes host RNA, RNA high-throughput sequencing library is constructed；
2.1. building is based on the DNA hybridization probe of host (mankind) cDNA library,
A) First-Strand cDNA Synthesis Using SuperScript is usedTM II RT(thermofisher
Scientific) by total serum IgE reverse transcription at cDNA, reaction system:
Following component is added in 65 DEG C of 5min:
25 DEG C, 2min, the SuperScript of 1 μ l (200units) is addedTM II RT。
25 DEG C of 10min, 42 DEG C of 50min, 72 DEG C of 15min.It can get the first chain product.
B) it with the RNA chain of RNase H enzyme (thermofisher scientific) digestion DNA-RNA compound, can obtain
To cDNA probe.Reaction system is as follows: 20 μ l, Rnase H of the first chain product, 0.2 μ l, 10X Rnase H buffer, 4 μ l,
Without RNA enzyme water 15.8ul.After reaction system mixes, 37 DEG C, 45min, RNA chain is removed, single-stranded cDNA is obtained.
C) single-stranded cDNA is carried out with the DNAClean XP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) of 1.5X
(covaris s220) is interrupted after purification and recovery and arrives 100bp or so, the DNA hybridization probe for hybridization can be obtained, and reacts item
Part is as follows:
Table 1: condition is interrupted
| Pipe | 50μl |
| Target BP (peak) | 200 |
| Highest incident power (W) | 175 |
| Service factor | 10% |
| Each pulse period | 200 |
| It handles time (s) | 1500 |
2.2.DNA hybridization probe is for host RNA removal in sample total serum IgE
DNA hybridization probe and sample total serum IgE are hybridized, system is as follows, 1 μ g of sample total serum IgE, DNA hybridization probe (2 μ g/ μ
L) 5 μ l, 10X SSC buffer, 5 μ l, no RNA enzyme water supply 50 μ l systems.Reaction condition: 95 DEG C of 5min 0.1 DEG C/s of cooling,
Drop to 4 DEG C.DNA:RNA heteroduplex can be obtained.
2.3. with the RNA chain of RNase H enzyme (thermofisher scientific) digestion DNA-RNA compound
Remove the host RNA in sample.
With remaining cDNA probe in Exonuclease I enzymatic treatment sample.0.5 μ L is added in above-mentioned product
Exonuclease I (50U/ μ L), reaction condition: 37 DEG C of 30min, 80 DEG C of 20min.
2.4 products are purified with the RNAClean XP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) of 2.2X
Recycling.Nonhost RNA product after concentration can be obtained.
A) 110 μ L (2.2X) RNACleanXP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) are taken, until 50 μ L samples
In this, piping and druming is mixed 10 times.
B) 15min is stood on ice, sample is placed in 5min on magnetic frame, after solution clarification, carefully removes supernatant.
C) 200 μ L80% ethyl alcohol are added and rinse magnetic bead, be incubated at room temperature 30s, carefully remove supernatant.
D) previous step is repeated.
E) exhaust liquid, and uncap dry 5-10min.
F) 10 μ L nuclease-free water are added, is blown and beaten 6 times and is mixed well with liquid-transfering gun, be stored at room temperature 2min.
G) sample, which is placed on magnetic frame, stands 5min, and after solution clarification, careful 8.5 μ L supernatants of drawing are to new
In nuclease-free PCR pipe.
3. reaction product KAPA RNA HyperPrep Kits kit constructs standard rna library, each sample is used
Nextseq500 sequenator tests 10Mreads.
4. analysis of biological information.
B: standard DNA testing process
1. extracting above-mentioned 2 parts of sample total-DNA using QIAamp DNA Blood Mini kit (QIAGEN).
2. interrupting program using Covaris s220 standard interrupts genomic DNA to 300bp.
3. preparing the DNA library of above-mentioned sample using NEBNext Ultra II DNA, each sample is used
Nextseq500 sequenator tests 10Mreads.
4. analysis of biological information.
2: two groups of methods and results of table compare
Illustrate the concentration effect of the RNA product for going host prepared according to the above method in upper table, it can be with by result
Find out:
1. 2 DNA cause of diseases can effectively detect DNA pathogen by the test of macro transcript profile；
2. 2 RNA cause of diseases can be detected effectively by the test of macro transcript profile；
3. the method for the present invention has preferable removal effect, the library host removed to host's rRNA and mRNA
The ratio of the total sequencing data of Zhan drastically reduces；
4. being better than DNA for our mixed DNA virus and DNA bacterium, RNA detection effect, it can be seen that the present invention
Method can achieve removal host RNA well, be enriched with the purpose of nonhost RNA.
Embodiment two: the method for the present invention detects unknown infection sample and clinical verification
Unknown infection type sample is tested, 5 Patients With Encephalitis cerebrospinal fluid (CSF) infection sample rnas is extracted and 1 mixed
There is the cerebrospinal fluid analog sample RNA of RNA virus (hepatitis C virus) and bacterium (Escherichia coli), constructs standard by the method for the present invention
Pathogen test situation is analyzed in the library RNA.
The method of the present invention testing process:
1. extracting above-mentioned 6 parts of sample total-RNA with RNeasy Mini Kit (Qiagen)；
2. host's probe cell using design removes host RNA, RNA high-throughput sequencing library is constructed；
2.1. building is based on the DNA hybridization probe of host (mankind) cDNA library,
A) First-Strand cDNA Synthesis Using SuperScript is usedTM II RT(thermofisher
Scientific) by total serum IgE reverse transcription at cDNA, reaction system:
Following component is added in 65 DEG C of 5min:
25 DEG C, 2min, the SuperScript of 1 μ L (200units) is addedTM II RT。
25 DEG C, 10min, 42 DEG C 50min, 72 DEG C of 15min.It can get the first chain product.
B) it with the RNA chain of RNase H enzyme (thermofisher scientific) digestion DNA-RNA compound, can obtain
To cDNA probe.Reaction system is as follows: 20 μ L, Rnase H of the first chain product, 0.2 μ L, 10X Rnase H buffer, 4 μ L,
Without 15.8 μ L of RNA enzyme water.After reaction system mixes, 37 DEG C, 45min, RNA chain is removed, single-stranded cDNA is obtained.
C) single-stranded cDNA is carried out with the DNAClean XP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) of 1.5X
(covaris s220) is interrupted after purification and recovery and arrives 100bp or so, the DNA hybridization probe for hybridization can be obtained, and reacts item
Part is as follows:
Table 1: condition is interrupted
| Pipe | 50μl |
| Target BP (peak) | 200 |
| Highest incident power (W) | 175 |
| Service factor | 10% |
| Each pulse period | 200 |
| It handles time (s) | 1500 |
2.2.DNA hybridization probe is for host RNA removal in sample total serum IgE
DNA hybridization probe and sample total serum IgE are hybridized, system is as follows, 1 μ g of sample total serum IgE, DNA hybridization probe (2 μ g/ μ
L) 5 μ l, 10X SSC buffer, 5 μ l, no RNA enzyme water supply 50 μ L systems.Reaction condition: 95 DEG C of 5min 0.1 DEG C/s of cooling,
Drop to 4 DEG C.DNA:RNA heteroduplex can be obtained.
2.3. with the RNA chain of RNase H enzyme (thermofisher scientific) digestion DNA-RNA compound
Remove the host RNA in sample.
With remaining cDNA probe in Exonuclease I enzymatic treatment sample.0.5 μ L is added in above-mentioned product
Exonuclease I (50U/ μ L), reaction condition: 37 DEG C of 30min, 80 DEG C, 20min.
2.4 products are purified with the RNAClean XP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) of 2.2X
Recycling.Nonhost RNA product after concentration can be obtained.
A) 110 μ L (2.2X) RNAClean XP Beads (Nanjing Nuo Weizan Bioisystech Co., Ltd) are taken, until 50 μ l samples
In this, piping and druming is mixed 10 times.
B) 15min is stood on ice, sample is placed in 5min on magnetic frame, after solution clarification, carefully removes supernatant.
C) 200 μ L, 80% ethyl alcohol is added and rinses magnetic bead, be incubated at room temperature 30s, carefully remove supernatant.
D) previous step is repeated.
E) exhaust liquid, and uncap dry 5-10min.
F) 10 μ L nuclease-free water are added, is blown and beaten 6 times and is mixed well with liquid-transfering gun, be stored at room temperature 2min.
G) sample, which is placed on magnetic frame, stands 5min, and after solution clarification, careful 8.5 μ L supernatants of drawing are to new
In nuclease-free PCR pipe.
3. reaction product KAPA RNA HyperPrep Kits kit constructs standard rna library, each sample is used
Nextseq500 sequenator tests 20Mreads.
4. analysis of biological information.
It is as follows to analyze result:
The number of each pathogen is special cause of disease sequence number (scaled value after uniform standardization) in upper table；In upper table
The detection effect for illustrating the RNA product of the encephalitis infection sample prepared according to the above method, can illustrate inhomogeneity by result
The disease-producing pathogens of type can be effectively detected, and almost the same with clinical diagnosis result；The RNA virus that we are prepared
It also can effectively be detected with DNA bacterium Quality Control immitation.It can be seen that the method for the present invention can achieve fine DNA and RNA detection
Effect.
It should be understood that the present invention disclosed is not limited only to specific method, scheme and the substance of description, because these
It is alterable.It will also be understood that purpose of the terminology used here just for the sake of the specific embodiment scheme of description, rather than
It is intended to limit the scope of the invention, the scope of the present invention is limited solely by the attached claims.
Those skilled in the art, which will also be appreciated that or be able to confirm that, uses no more than routine experiment, institute herein
The many equivalents for the specific embodiment of the invention stated.These equivalents are also contained in the attached claims.
Claims (10)
1. a kind of method that detection pathogen is sequenced by macro transcript profile, which is characterized in that the method passes through only detection RNA mould
Formula is come while detecting DNA and RNA pathogen.
2. the method according to claim 1, wherein the described method comprises the following steps:
Step 1 prepares total-RNA sample from biological sample to be measured；
Step 2 removes host RNA using host's probe cell of design, constructs RNA high-throughput sequencing library；
Step 3 carries out high-flux sequence using the RNA high-throughput sequencing library, obtains RNA sequencing data；
Step 4 carries out bioinformatic analysis to the RNA sequencing data, it is to be checked to determine whether the biological sample to be measured contains
Survey pathogen.
3. according to the method described in claim 2, it is characterized in that, the biological sample to be measured be selected from peripheral blood, lesion tissue,
Cerebrospinal fluid, alveolar wash one's hands and face liquid, Pleural effusions, sputum, respiratory secretions or digestive tract secretion.
4. according to the method described in claim 2, it is characterized in that, the high-flux sequence uses illumina Nextseq
500 sequenators are sequenced.
5. according to the method described in claim 2, it is characterized in that, removing host RNA in the step 2 includes conservative ribose
Body RNA, globulin mRNA and host's height express mRNA.
6. according to the method described in claim 5, it is characterized in that, the conservative rRNA includes: mitochondria rRNA 16s
RRNA, 12s rRNA；Cytoplasm rRNA 28s rRNA, 5.8s rRNA, 18s rRNA, 5s rRNA.
7. according to the method described in claim 2, it is characterized in that, the step 4 the following steps are included:
A. Adapter, Duplication, Poly structure are removed from the RNA sequencing data and filter quality value is lower than Q20
Reads, removal length be less than 50bp reads, obtain valid data；
B. the valid data are compared with host genome data, the reads compared therewith is removed, is counted
Host data amount retains nonhost data, obtains and compares to the reads quantity N on hosthWith host genome length Lh；
C. the nonhost data are compared with the property data base of pathogen to be detected, obtain representative and compares to be detected
Reads quantity, pathogen N to be detected on pathogen iiGenome length Li, comparison result is screened, determines the life to be measured
Whether object sample contains pathogen to be detected.
8. the method according to the description of claim 7 is characterized in that if the biological sample contains pathogen to be detected, by institute
It states cause of disease data to be standardized using following calculation formula, determines pathogen detection value Ti,
。
9. the method according to the description of claim 7 is characterized in that the host genome data are full-length genome data；Institute
It states valid data and is compared with host genome data by bowtie bwa software.
10. the method according to claim 1, wherein the pathogen to be detected be helminth, fungi, bacterium,
Actinomyces, virus, mycoplasma, Chlamydia, Richettsia or conveyor screw.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910652791.9A CN110317864A (en) | 2019-07-18 | 2019-07-18 | A method of it is sequenced by macro transcript profile to detect pathogen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910652791.9A CN110317864A (en) | 2019-07-18 | 2019-07-18 | A method of it is sequenced by macro transcript profile to detect pathogen |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110317864A true CN110317864A (en) | 2019-10-11 |
Family
ID=68124110
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910652791.9A Pending CN110317864A (en) | 2019-07-18 | 2019-07-18 | A method of it is sequenced by macro transcript profile to detect pathogen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110317864A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113517025A (en) * | 2020-04-10 | 2021-10-19 | 西咸新区予果微码生物科技有限公司 | Pathogen online monitoring system and method |
| CN114107454A (en) * | 2020-08-28 | 2022-03-01 | 上海交通大学医学院附属上海儿童医学中心 | Respiratory tract infection pathogen detection method based on macrogene/macrotranscriptome sequencing |
| CN116949154A (en) * | 2023-08-28 | 2023-10-27 | 中国农业大学 | Macrotranscriptome-based pathogen detection method for non-therapeutic purpose |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105112569A (en) * | 2015-09-14 | 2015-12-02 | 中国医学科学院病原生物学研究所 | Virus infection detection and identification method based on metagenomics |
| CN105224824A (en) * | 2015-09-28 | 2016-01-06 | 山东出入境检验检疫局检验检疫技术中心 | Based on the duck tembusu virus nondiagnostic detection method of metagenomics |
| US20160304953A1 (en) * | 2013-12-04 | 2016-10-20 | University Of Alaska Fairbanks | Methods and compositions for enriching non-host sequences in host samples |
| CN108660199A (en) * | 2018-05-20 | 2018-10-16 | 北京宏微特斯生物科技有限公司 | A method of pathogen is detected based on cfDNA high-flux sequences |
-
2019
- 2019-07-18 CN CN201910652791.9A patent/CN110317864A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20160304953A1 (en) * | 2013-12-04 | 2016-10-20 | University Of Alaska Fairbanks | Methods and compositions for enriching non-host sequences in host samples |
| CN105112569A (en) * | 2015-09-14 | 2015-12-02 | 中国医学科学院病原生物学研究所 | Virus infection detection and identification method based on metagenomics |
| CN105224824A (en) * | 2015-09-28 | 2016-01-06 | 山东出入境检验检疫局检验检疫技术中心 | Based on the duck tembusu virus nondiagnostic detection method of metagenomics |
| CN108660199A (en) * | 2018-05-20 | 2018-10-16 | 北京宏微特斯生物科技有限公司 | A method of pathogen is detected based on cfDNA high-flux sequences |
Non-Patent Citations (2)
| Title |
|---|
| 李定辰: "基于高通量测序平台的未知病原微生物检测系统", 《中国博士学位论文全文数据库》 * |
| 王曦等: "新一代高通量RNA测序数据的处理与分析", 《生物化学与生物物理进展》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113517025A (en) * | 2020-04-10 | 2021-10-19 | 西咸新区予果微码生物科技有限公司 | Pathogen online monitoring system and method |
| CN113517025B (en) * | 2020-04-10 | 2022-03-18 | 西咸新区予果微码生物科技有限公司 | Pathogen online monitoring system and method |
| CN114107454A (en) * | 2020-08-28 | 2022-03-01 | 上海交通大学医学院附属上海儿童医学中心 | Respiratory tract infection pathogen detection method based on macrogene/macrotranscriptome sequencing |
| CN116949154A (en) * | 2023-08-28 | 2023-10-27 | 中国农业大学 | Macrotranscriptome-based pathogen detection method for non-therapeutic purpose |
| CN116949154B (en) * | 2023-08-28 | 2024-05-28 | 中国农业大学 | Macrotranscriptome-based pathogen detection method for non-therapeutic purpose |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2699666B1 (en) | miRNA-BASED UNIVERSAL SCREENING TEST (UST) | |
| CN110317864A (en) | A method of it is sequenced by macro transcript profile to detect pathogen | |
| CN107660234A (en) | The method of prediction organ-graft refection is sequenced using two generations | |
| CN107312839B (en) | Substance, kit and method for detecting systemic infection | |
| CN111500781B (en) | Amplification primer group for detecting SARS-CoV-2 by mNGS and application thereof | |
| CN103060309B (en) | Extraction method for metagenome | |
| CN112501268A (en) | Nanopore sequencing-based primer group and kit for rapidly identifying respiratory microorganisms and application of primer group and kit | |
| CN111154754A (en) | Probe set for analyzing DNA sample and method for using the same | |
| CN107893260A (en) | Efficiently remove the method and kit of the structure transcript profile sequencing library of rRNA | |
| CN110387438A (en) | Multi-primers, kit and method for enterovirus high-flux sequence | |
| CN110747269A (en) | Granular cell biomarker for PCOS diagnosis, screening method thereof and diagnostic kit | |
| CN112725410A (en) | Primer group for detecting pathogenic microorganisms | |
| WO2019085546A1 (en) | Method for constructing microbial 16s rdna single-molecule level sequencing library | |
| CN110157808A (en) | A kind of application of non-coding RNA relevant to larynx squamous carcinoma occurrence and development | |
| US8178343B2 (en) | Gene cluster diagnosis apparatus | |
| CN109837344B (en) | Methylated EphA7 nucleotide fragment, detection method and application thereof | |
| Li et al. | Characterization of MicroRNA cargo of extracellular vesicles isolated from the plasma of Schistosoma japonicum-infected mice | |
| Chen et al. | The screening and validation process of miR-223-3p for saliva identification | |
| CN109517819A (en) | A kind of detection probe, method and kit modified for detecting multiple target point gene mutation, methylation modification and/or methylolation | |
| CN109628585A (en) | Application of the non-coding RNA in diagnosis of sepsis disease | |
| CN109811052A (en) | A kind of kit and gene panel detecting idiopathic azoospermatism | |
| CN109504807B (en) | Nucleic acid sequence combination for detecting herpes virus type 4 and application thereof | |
| CN105176992B (en) | The micro-deleted detection primer in the areas Y chromosome AZF | |
| CN118291603A (en) | Method for predicting biological age of individual, detection reagent and kit | |
| CN105506105B (en) | Kit for detecting proliferation capacity and clinical application value of hUCMSCs |
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 |