CN107058622A - A kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen - Google Patents
A kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen Download PDFInfo
- Publication number
- CN107058622A CN107058622A CN201710201003.5A CN201710201003A CN107058622A CN 107058622 A CN107058622 A CN 107058622A CN 201710201003 A CN201710201003 A CN 201710201003A CN 107058622 A CN107058622 A CN 107058622A
- Authority
- CN
- China
- Prior art keywords
- probe
- specific primer
- kit
- reaction solution
- pcr method
- 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.)
- Granted
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
- C12Q1/70—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
- C12Q1/686—Polymerase chain reaction [PCR]
-
- 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
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- 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
- C12Q2600/16—Primer sets for multiplex assays
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Virology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen, includes 6 components：Reaction solution A, reaction solution B, reaction liquid C, enzyme mixation, positive control, negative control are detected comprising 11 kinds of common respiratory pathogens（Influenza A virus is universal, influenza B virus, Respiratory Syncytial Virus(RSV), 1/2/3 type of human parainfluenza viruses, adenovirus, mycoplasma pneumoniae, CPN, legionella pneumophilia, streptococcus pneumonia, haemophilus influenzae, A races streptococcus）, expanded by 3 reaction buffers, each reaction buffer includes four fluorescence channels, can investigate clinically 90% pathogenic infection.
Description
Technical field
The invention belongs to quantitative fluorescent PCR field, and in particular to a kind of multiple fluorescence PCR method joint-detection respiratory pathogenses
The kit of body.
Background technology
ARI has higher morbidity and mortality, particularly ALRI to cause great disease
Burden, the preschool child that the whole world has 20% dies from ALRI（SARI）, 90% death is attributed to pneumonia.Pneumonia
It is also to cause children to fall ill and main causes of death or acute pneumonia is the main infection form of heating respiratory tract symptom grouping.
Within 1 year old the children Streptococcus incidence of disease be 0.01 ~ 0.68 time/man-year, within 5 years old the children Streptococcus incidence of disease be 0.14 ~
0.66 time/man-year；Pneumonia death rate children below 1 years old are 485/100,000 ~ 890/100,000, and less than 5 years old children are 184/10
Ten thousand ~ 1 223/100,000.（Data source refers to " China's Mainland pneumonia incidence and mortality "）.
At this stage clinically to respiratory pathogen mainly by clinical symptoms, be separately cultured and the conventional hand such as immune detection
Duan Jinhang is diagnosed and pathogen investigation.It is separately cultured and takes time and effort, it is necessary to strict laboratory environment, usually require 3 days ability
Make a definite diagnosis；And sensitivity of immune detection is low, and there is " window phase ", can just detect corresponding antigen or antibody within 3 ~ 7 days after infection.
The content of the invention
It is main it is an object of the invention to provide a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen
Multiple real time fluorescence PCR methods are taken to detect respiratory pathogen.
To achieve the above object, the present invention is adopted the following technical scheme that：
The present invention is detected in a kit comprising 11 kinds of common respiratory pathogens（Influenza A virus is universal, second
Type influenza virus, Respiratory Syncytial Virus(RSV), 1/2/3 type of human parainfluenza viruses, adenovirus, mycoplasma pneumoniae, CPN,
Legionella pneumophilia, streptococcus pneumonia, haemophilus influenzae, A races streptococcus）, expanded by 3 reaction buffers, each
Reaction buffer includes four fluorescence channels, can investigate clinically 90% pathogenic infection.
The kit includes as follows；Reaction solution A includes Flu-A specific primer and probe, influenza B specificity
Primer and probe, adenovirus specific primer and probe, Respiratory Syncytial Virus(RSV) specific primer and probe；
Reaction solution B includes mycoplasma pneumoniae specific primer and probe, CPN specific primer and probe, parainfluenza virus
Malicious specific primer and probe, internal control specific primer and probe；
Reaction liquid C is bloodthirsty comprising legionella pneumophilia specific primer and probe, streptococcus pneumonia specific primer and probe, influenza
Bacillus specific primer and probe, A races streptococcus specific primer and probe；
Enzyme mixation includes hot start Taq polymerase and M-MLV enzymes；
Positive control includes specific plasmids and TE water；
Negative control includes TE water.
Primer probe sequence：
Reagent component is formulated：
Reaction solution A is formulated：Tris（PH8.8）20mmol/L, KCl 60mmol/L, MgCl2 2.5mmol/L, EDTA2Na
0.1mmol/L, 1%DMSO, dATP 200 μm of ol/L, dGTP 200 μm of ol/L, dCTP 200 μm of ol/L of 200 μm of ol/L, dTTP,
Influenza A virus sense primer 417nmol/L, influenza A virus anti-sense primer 417nmol/L, influenza A probe
125nmol/L, influenza B virus sense primer 417nmol/L, influenza B virus anti-sense primer 417nmol/L, B-mode stream
Influenza Virus probe 125nmol/L, adenovirus sense primer 417nmol/L, adenovirus anti-sense primer 417nmol/L, adenovirus are visited
Pin 125nmol/L, Respiratory Syncytial Virus(RSV) sense primer 417nmol/L, Respiratory Syncytial Virus(RSV) anti-sense primer 417nmol/L,
Respiratory Syncytial Virus(RSV) probe 125nmol/L.
Reaction solution B is formulated：Tris（PH8.8）20mmol/L, KCl 60mmol/L, MgCl2 2.5mmol/L, EDTA2Na
0.1mmol/L, 1%DMSO, dATP 200 μm of ol/L, dGTP 200 μm of ol/L, dCTP 200 μm of ol/L of 200 μm of ol/L, dTTP,
Mycoplasma pneumoniae sense primer 417nmol/L, mycoplasma pneumoniae anti-sense primer 417nmol/L, mycoplasma pneumoniae probe
125nmol/L, CPN sense primer 417nmol/L, CPN anti-sense primer 417nmol/L, CPN
Probe 125nmol/L, parainfluenza virus sense primer 417nmol/L, parainfluenza virus anti-sense primer 417nmol/L, parainfluenza
Viral Probe 125nmol/L, internal control sense primer 417nmol/L, internal control anti-sense primer 417nmol/L, internal control probe
125nmol/L。
Reaction liquid C is formulated：Tris（PH8.8）20mmol/L, KCl 60mmol/L, MgCl2 2.5mmol/L, EDTA2Na
0.1mmol/L, 1%DMSO, dATP 200 μm of ol/L, dGTP 200 μm of ol/L, dCTP 200 μm of ol/L of 200 μm of ol/L, dTTP,
Streptococcus pneumonia sense primer 417nmol/L, streptococcus pneumonia anti-sense primer 417nmol/L, streptococcus pneumonia probe
125nmol/L, legionella pneumophilia sense primer 417nmol/L, legionella pneumophilia anti-sense primer 417nmol/L, legionella pneumophilia
Streptococcus anti-sense primer 417nmol/L, A races of streptococcus sense primer 417nmol/L, A races of probe 125nmol/L, A races streptococcus
Probe 125nmol/L, haemophilus influenzae sense primer 417nmol/L, haemophilus influenzae anti-sense primer 417nmol/L, stream
Haemophilus influenza probe 125nmol/L.
Enzyme mixation is formulated：Taq enzyme 4U/ μ L, M-MLV enzyme 10U/ μ L.
The advantage of the invention is that：
The advantage of invention：The advantage of the present invention is 1, sensitivity reaches 500copies/mL；2nd, result is directly obtained after having expanded,
Without subsequent operation, 3 hours are obtained after sample with regard to result can be obtained；3rd, the without window phase, once infection with regard to that can detect.
The beneficial effect of invention：1st, detection sensitivity is improved, increases recall rate, shortens the course of disease of patient；2nd, the clear and definite cause of disease,
Improve medication accuracy rate, it is to avoid drug abuse；3rd, manual operations is reduced, shortens testing process, testing cost is reduced.
Brief description of the drawings
Fig. 1 reaction solution A specific outcome figures.
Fig. 2 reaction solution B specific outcome figures.
Fig. 3 reaction liquid C specific outcome figures.
Fig. 4 reaction solution A linear sensitivity result figures.
Fig. 5 reaction solution B linear sensitivity result figures.
Fig. 6 reaction liquid C linear sensitivity result figures.
Embodiment
Embodiment 1
1st, the preparation of reaction solution
（1）Reaction solution A preparation
10mL volumetric flasks are taken, 0.5mol/L Trizma are separately added into®HCl 64 μ L, 0.5mol/L Trizma® Base 336μ
L, 1 mol/L MgCl225 μ L, 1mol/L KCl, 600 μ L, 0.5mol/L EDTA2Na 2 μ L, DMSO 100 μ L, dNTPs
90 μ L, 50 μm of ol/L influenza A virus upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/L influenza A probes 25,
50 μm of ol/L influenza B virus upstream and downstream primer each 83.4 μ L, 50 μm of ol/L influenza B virus probes 25 μ L, 50 μm of ol/
In L adenovirus upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/L adenovirus probe 25,50 μm of ol/L Respiratory Syncytial Virus(RSV)
Anti-sense primer each 83.4 μ L, 50 μm of μ L of ol/L Respiratory Syncytial Virus(RSV) probe 25.Volume is supplied to 10mL with dual distilled water, is turned over
Turn to be allowed to fully mixing, liquid is transferred in 10mL beakers, be dispensed into by 1mL/ branch in centrifuge tube, -20 DEG C save backup.
（2）Reaction solution B preparation
10mL volumetric flasks are taken, 0.5mol/L Trizma are separately added into®HCl 64 μ L, 0.5mol/L Trizma® Base 336μ
L, 1 mol/L MgCl225 μ L, 1mol/L KCl, 600 μ L, 0.5mol/L EDTA2Na 2 μ L, DMSO 100 μ L, dNTPs
90 μ L, 50 μm of ol/L mycoplasma pneumoniae upstream and downstream primer each 83.4 μ L, 50 μm of ol/L mycoplasma pneumoniaes probes 25 μ L, 50 μ
Mol/L CPN upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/L CPNs probe 25,50 μm of ol/L sidestream
Influenza Virus upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/L parainfluenza viruses probe 25,50 μm of ol/L internal control upstream and downstream primer
Each 83.4 μ L, 50 μm of μ L of ol/L internal controls probe 25.Volume is supplied with dual distilled water to 10mL, upset is allowed to fully mixing, will
Liquid is transferred in 10mL beakers, is dispensed into by 1mL/ branch in centrifuge tube, -20 DEG C save backup.
（3）The preparation of reaction liquid C
10mL volumetric flasks are taken, 0.5mol/L Trizma are separately added into®HCl 64 μ L, 0.5mol/L Trizma® Base 336μ
L, 1 mol/L MgCl225 μ L, 1mol/L KCl, 600 μ L, 0.5mol/L EDTA2Na 2 μ L, DMSO 100 μ L, dNTPs
90 μ L, 50 μm of ol/L streptococcus pneumonia upstream and downstream primer each 83.4 μ L, 50 μm of ol/L streptococcus pneumonias probes 25 μ L, 50 μ
Mol/L legionella pneumophilia upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/L legionella pneumophilias probe 25,50 μm of ol/L A races
Above and below streptococcus upstream and downstream primer each 83.4 μ L, 50 μm of μ L of ol/LA races streptococcus probe 25,50 μm of ol/L haemophilus influenzae
Swim primer each 83.4 μ L, 50 μm of μ L of ol/L haemophilus influenzaes probe 25.Volume is supplied with dual distilled water to 10mL, upset makes
Abundant mixing, liquid is transferred in 10mL beakers, is dispensed into by 1mL/ branch in centrifuge tube, -20 DEG C save backup.
（4）The preparation of enzyme mixation
10mL volumetric flasks are taken, 10000U/mL Taq enzyme 4mL, 25000U/mL M-MLV enzymes 4mL is separately added into.Use dual distillation
Water supplies volume to 10mL, and upset is allowed to fully mixing, liquid is transferred in 10mL beakers, and centrifugation is dispensed into by 100 μ L/ branch
Guan Zhong, -20 DEG C save backup.
（5）The preparation of negative control
100mL volumetric flasks are taken, physiological saline is added and is settled to 100mL, liquid is transferred in 10mL beakers, by 500 μ L/ branch point
It is attached in centrifuge tube, -20 DEG C save backup.
（6）The preparation of positive control（Concentration is 5.0 × 103copies/mL）
100mL volumetric flasks are taken, 5.0 × 10 are separately added into7Copies/mL influenza A virus, influenza B virus, adenovirus,
Respiratory Syncytial Virus(RSV), mycoplasma pneumoniae, CPN, parainfluenza virus, streptococcus pneumonia, legionella pneumophilia, A races hammer
Bacterium and haemophilus influenzae plasmid（Each plasmid entrusts general biosystem（Anhui）Co., Ltd synthesizes）Each 10 μ L.Use physiology salt
Water is settled to 100mL, and liquid is transferred in 100mL beakers, is dispensed into by 500 μ L/ branch in centrifuge tube, -20 DEG C save backup.
2nd, nucleic acid extraction
Nucleic acid extraction is carried out using the nucleic acid extraction kit put on record, it is pure with micro ultraviolet specrophotometer survey nucleic acid after extraction
Degree, its OD260/OD280 should be between 1.6 ~ 2.0.
3rd, machine on sample-adding
（1）Reactant mixture is prepared
Take out reaction solution A, reaction solution B, reaction liquid C, enzyme mixation room temperature to place, it is fully dissolved, prepare reactant mixture
（Per test configurations system：The μ L enzyme mixations of 29.5 μ L reaction solutions+0.5）, reagent dosage is calculated as required, is sufficiently mixed
After uniform, 3000-5000g is centrifuged 5 seconds.
（2）Sample-adding
0.2mL amplifications pipe or eight unions are taken, the reactant mixture that 30 μ L are prepared is added, the sample extracted is then taken into 2 μ
L adds amplification pipe or eight unions, covers lid, is inserted after somewhat centrifuging in fluorescent PCR amplification instrument.
（3）Upper machine testing
1）Cycling condition is set
38 DEG C 5 minutes, 95 DEG C 10 minutes；Into following circulation：95 DEG C 15 seconds, 58 DEG C 40 seconds（Detect signal）, 40 circulations；
25 DEG C 30 seconds.
2）Instrument sense channel is selected
Fluorescein is set as FAM, ROX, HEX and CY5.
4th, result judges
（1）Negative control should be without Ct values or for 0；Positive control Ct values answer≤36；CY5 channel Cs t values answer≤36 in reaction solution B；
（2）Sample test result answers following table to judge
Embodiment 2
1st, reagent specificity verification
（1）Experiment sample
Take 10 parts of specific samples to verify the specificity of reagent, wherein 4 parts be physiological saline, 1 part be the inclined tuberculosis of people
Malicious sample, 1 part be rubella virus sample, 1 part be measles virus sample, 1 part be Friedlander's bacillus sample, 1 part be large intestine
Bacillus sample, 1 part be staphylococcus aureus sample.
（2）Experimentation
With reaction solution A, reaction solution B, respectively reaction liquid C, more than detection 10 parts of specific samples, testing result, checking examination are analyzed
The specificity of agent.
（3）Experimental result
Three kinds of reaction solutions detect that 10 parts of specific samples are all negative, show that reagent specificity is good, no cross reaction situation.Specifically
As a result it see the table below：
Reaction solution A specific detection results
Reaction solution B specific detection results
Reaction liquid C specific detection result
2nd, reagent accurate is verified
（1）Experiment sample
Take 1 part of accuracy sample（Concentration is 5.0 × 103copies/mL）Accuracy to reagent verifies that sample is prepared
Method is separately added into 5.0 × 10 to take 100mL volumetric flasks7Copies/mL influenza A virus, influenza B virus, adenopathy
Poison, Respiratory Syncytial Virus(RSV), mycoplasma pneumoniae, CPN, parainfluenza virus, internal control, streptococcus pneumonia, Shi Fei legions
Bacterium, A races streptococcus and each 10 μ L of haemophilus influenzae plasmid, 100mL is settled to physiological saline.
（2）Experimentation
With reaction solution A, reaction solution B, respectively reaction liquid C, repeatedly detection above accuracy sample 10 times, testing result is analyzed, is tested
Demonstrate,prove the accuracy of reagent.
（3）Experimental result
Three kinds of reaction solutions detect accuracy sample variation within batch coefficient（CV values）Equal ＜ 5%, shows that reagent is reproducible.Specific knot
Fruit see the table below：
Reaction solution A accuracy testing results
Reaction solution B accuracy testing results
Reaction liquid C accuracy testing result
3rd, reagent minimum detection limit is verified
（1）Experiment sample
Take 1 part of minimum detection limit sample（Concentration is 5.0 × 102copies/mL）Minimum detection limit to reagent verifies,
Sample compound method is separately added into 5.0 × 10 to take 100mL volumetric flasks3Copies/mL accuracy sample 10mL, uses physiology
Salt solution is settled to 100mL.
（2）Experimentation
With reaction solution A, reaction solution B, reaction liquid C, respectively repeatedly detection above minimum detection limit sample 10 times, analysis detection knot
Really, the minimum detection limit of reagent is verified.
（3）Experimental result
Three kinds of reaction solution detection minimum detection limit samples are the positive, and the lowest detection of reagent is limited to 5.0 × 102copies/mL。
Concrete outcome see the table below：
4th, reagent linear sensitivity is verified
（1）Experiment sample
Take 8 parts of line style sensitivity samples（Concentration is respectively 5.0 × 108copies/mL、5.0×107copies/mL、5.0×
106copies/mL、5.0×105copies/mL、5.0×104copies/mL、5.0×103copies/mL、5.0×
102copies/mL、5.0×101copies/mL、）The linear of reagent is verified.
（2）Experimentation
With reaction solution A, reaction solution B, respectively reaction liquid C, detection above linear sample, testing result is analyzed, the line of reagent is verified
Intelligent sensitivity.
（3）Experimental result
Three kinds of reaction solutions detection line style sensitivity samples are the positive, reagent it is linear good.Concrete outcome See Figure 4-6.
Embodiment 3：Detect the result of 120 clinical samples
1st, according to the compound method shown in embodiment 1, reagent preparation box related component is saved backup in -20 DEG C.
2nd, healthcare hospital for women ＆ children of Wuhan City have collected 120 have clinical respiratory infection symptoms patient's throat swab sample and
Sputum sample, must Acer biotechnology using moral（Xiamen）The nucleic acid extracting reagent of Co., Ltd's production（Virus type）Nucleic acid is carried out to carry
Take, with the purity of UV spectrophotometer measuring DNA sample, 120 sample OD260/OD280 are all between 1.6 ~ 2.0.
3rd, according to the step shown in embodiment 1, DNA sample-addings is carried out and upper quantitative real time PCR Instrument is detected that this makes
Instrument is ABI7500.
4th, according to criterion shown in embodiment 1, interpretation is carried out to result and is counted, as a result such as following table：
Presently preferred embodiments of the present invention is the foregoing is only, all equivalent changes done according to scope of the present invention patent are with repairing
Decorations, should all belong to the covering scope of the present invention.
SEQUENCE LISTING
<110>Moral must Acer biotechnology (Xiamen) Co., Ltd
<120>A kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen
<130> 36
<160> 36
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<400> 1
tgatcctctc gtcattgcag 20
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
<400> 2
ctcaggcact ccttccgtag 20
<210> 3
<211> 26
<212> DNA
<213>Artificial sequence
<400> 3
tgggatcttg cacctgatat tgtgga 26
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence
<400> 4
atgcaagggt ttccatgttc 20
<210> 5
<211> 20
<212> DNA
<213>Artificial sequence
<400> 5
accctccgtc tccacctact 20
<210> 6
<211> 21
<212> DNA
<213>Artificial sequence
<400> 6
tccgatgacc agatctgggg g 21
<210> 7
<211> 20
<212> DNA
<213>Artificial sequence
<400> 7
agtcttgcat gagccgttct 20
<210> 8
<211> 20
<212> DNA
<213>Artificial sequence
<400> 8
ggactcacac gtatgcatgg 20
<210> 9
<211> 21
<212> DNA
<213>Artificial sequence
<400> 9
cgggcacttc ttcctcaccc g 21
<210> 10
<211> 20
<212> DNA
<213>Artificial sequence
<400> 10
gtgcagggca agtgatgtta 20
<210> 11
<211> 20
<212> DNA
<213>Artificial sequence
<400> 11
tgatgctttt gggttgttca 20
<210> 12
<211> 25
<212> DNA
<213>Artificial sequence
<400> 12
tgctcagaaa ttgggtggag aagca 25
<210> 13
<211> 20
<212> DNA
<213>Artificial sequence
<400> 13
cagctcgtgt cgtgagatgt 20
<210> 14
<211> 20
<212> DNA
<213>Artificial sequence
<400> 14
ttgacgtcat cccttccttc 20
<210> 15
<211> 20
<212> DNA
<213>Artificial sequence
<400> 15
agtcccgcaa cgagcgcaac 20
<210> 16
<211> 20
<212> DNA
<213>Artificial sequence
<400> 16
agccataacg ccgtgaatac 20
<210> 17
<211> 20
<212> DNA
<213>Artificial sequence
<400> 17
atcccagtca tcagcctcac 20
<210> 18
<211> 20
<212> DNA
<213>Artificial sequence
<400> 18
cccgggcctt gtacacaccg 20
<210> 19
<211> 20
<212> DNA
<213>Artificial sequence
<400> 19
caggtgtcac ggctgttcta 20
<210> 20
<211> 20
<212> DNA
<213>Artificial sequence
<400> 20
ttccgtctgg gttttactgg 20
<210> 21
<211> 23
<212> DNA
<213>Artificial sequence
<400> 21
ccaatgcagc agaggcaaaa tcc 23
<210> 22
<211> 20
<212> DNA
<213>Artificial sequence
<400> 22
ttatcactgg cggaaagacc 20
<210> 23
<211> 20
<212> DNA
<213>Artificial sequence
<400> 23
tcagttcaac cgctgcatag 20
<210> 24
<211> 20
<212> DNA
<213>Artificial sequence
<400> 24
acgttggggg cggttggaat 20
<210> 25
<211> 20
<212> DNA
<213>Artificial sequence
<400> 25
ggctttaacc gaacagcaaa 20
<210> 26
<211> 20
<212> DNA
<213>Artificial sequence
<400> 26
ttgcaaacca cttggcaata 20
<210> 27
<211> 23
<212> DNA
<213>Artificial sequence
<400> 27
caaaaacaag ccaggcgttg ttg 23
<210> 28
<211> 20
<212> DNA
<213>Artificial sequence
<400> 28
gtgttttcgg cacaaaaggt 20
<210> 29
<211> 20
<212> DNA
<213>Artificial sequence
<400> 29
cgcactaaac ccttcagctc 20
<210> 30
<211> 20
<212> DNA
<213>Artificial sequence
<400> 30
cagctatgcg gcgtgcctca 20
<210> 31
<211> 20
<212> DNA
<213>Artificial sequence
<400> 31
ccgacttgca tttgctctct 20
<210> 32
<211> 20
<212> DNA
<213>Artificial sequence
<400> 32
gaacggtctt tgcgtttctc 20
<210> 33
<211> 21
<212> DNA
<213>Artificial sequence
<400> 33
aggggattcg cgctttgcag a 21
<210> 34
<211> 20
<212> DNA
<213>Artificial sequence
<400> 34
acagtcagcc gcatcttctt 20
<210> 35
<211> 20
<212> DNA
<213>Artificial sequence
<400> 35
acgaccaaat ccgttgactc 20
<210> 36
<211> 20
<212> DNA
<213>Artificial sequence
<400> 36
cagccgagcc acatcgctca 20
Claims (5)
1. a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen, it is characterised in that；The kit includes
It is as follows：Reaction solution A is special comprising Flu-A specific primer and probe, influenza B specific primer and probe, adenovirus
Property primer and probe, Respiratory Syncytial Virus(RSV) specific primer and probe；
Reaction solution B includes mycoplasma pneumoniae specific primer and probe, CPN specific primer and probe, parainfluenza virus
Malicious specific primer and probe, internal control specific primer and probe；
Reaction liquid C is bloodthirsty comprising legionella pneumophilia specific primer and probe, streptococcus pneumonia specific primer and probe, influenza
Bacillus specific primer and probe, A races streptococcus specific primer and probe.
2. a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen according to claim 1, its feature
It is：The kit also includes as follows：Enzyme mixation includes hot start Taq polymerase and M-MLV enzymes；
Positive control includes specific plasmids and TE water；
Negative control includes TE water.
3. a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen according to claim 1, its feature
It is；Flu-A specific primer is in described reaction solution A：F：tgatcctctcgtcattgcag、R：
Ctcaggcactccttccgtag, probe is FAM-tgggatcttgcacctgatattgtgga-BHQ1；
Influenza B specific primer is：F：atgcaagggtttccatgttc、R：Accctccgtctccacctact, probe is
HEX-tccgatgaccagatctggggg-BHQ1；
Adenovirus specific primer is：F：agtcttgcatgagccgttct、R：Ggactcacacgtatgcatgg, probe is
ROX-cgggcacttcttcctcacccg-BHQ2；
Respiratory Syncytial Virus(RSV) specific primer is：F：gtgcagggcaagtgatgtta、R：Tgatgcttttgggttgttca,
Probe is CY5-tgctcagaaattgggtggagaagcaBHQ2.
4. a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen according to claim 1, its feature
It is；Mycoplasma pneumoniae specific primer is in described reaction solution B：F：cagctcgtgtcgtgagatgt、R：
Ttgacgtcatcccttccttc, probe is FAM-agtcccgcaacgagcgcaac-BHQ1；
CPN specific primer is：F：agccataacgccgtgaatac、R：Atcccagtcatcagcctcac, probe
For HEX-cccgggccttgtacacaccg-BHQ1；
Parainfluenza virus specific primer is：F：caggtgtcacggctgttcta、R：Ttccgtctgggttttactgg, probe
For ROX-ccaatgcagcagaggcaaaatcc-BHQ2；
Internal control specific primer is：F：acagtcagccgcatcttctt、R：Acgaccaaatccgttgactc, probe is CY5-
cagccgagccacatcgctca-BHQ2。
5. a kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen according to claim 1, its feature
It is；Legionella pneumophilia specific primer is in described reaction liquid C：F：ggctttaaccgaacagcaaa、R：
Ttgcaaaccacttggcaata, probe is HEX-caaaaacaagccaggcgttgttg-BHQ1；
Streptococcus pneumonia specific primer is：F：ttatcactggcggaaagacc、R：Tcagttcaaccgctgcatag, probe
For FAM-acgttgggggcggttggaat-BHQ1；
Haemophilus influenzae specific primer is：F：ccgacttgcatttgctctct、R：Gaacggtctttgcgtttctc, is visited
Pin is CY5-aggggattcgcgctttgcaga-BHQ2；
A races streptococcus specific primer is：F：gtgttttcggcacaaaaggt、R：Cgcactaaacccttcagctc, probe
For ROX-cagctatgcggcgtgcctca-BHQ2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710201003.5A CN107058622B (en) | 2017-03-30 | 2017-03-30 | Kit for joint detection of respiratory pathogens by multiple fluorescence PCR method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710201003.5A CN107058622B (en) | 2017-03-30 | 2017-03-30 | Kit for joint detection of respiratory pathogens by multiple fluorescence PCR method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107058622A true CN107058622A (en) | 2017-08-18 |
| CN107058622B CN107058622B (en) | 2020-03-27 |
Family
ID=59601744
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710201003.5A Active CN107058622B (en) | 2017-03-30 | 2017-03-30 | Kit for joint detection of respiratory pathogens by multiple fluorescence PCR method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107058622B (en) |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630098A (en) * | 2017-11-13 | 2018-01-26 | 湖南圣湘生物科技有限公司 | Fluorescent PCR detection architecture, kit and detection method for joint-detection various respiratory road bacterium |
| CN107828633A (en) * | 2017-12-14 | 2018-03-23 | 苏州天隆生物科技有限公司 | A kind of nucleic acid extraction and reagent device, reagent and the method for detection |
| CN108374059A (en) * | 2018-03-01 | 2018-08-07 | 绍兴迅敏康生物科技有限公司 | Respiratory Syncytial Virus(RSV) kit for detecting nucleic acid and its detection method |
| CN109609692A (en) * | 2018-12-25 | 2019-04-12 | 北京卓诚惠生生物科技股份有限公司 | For detecting the nucleic acid reagent, kit, system and method for respiratory tract infection pathogen |
| CN109897917A (en) * | 2019-04-01 | 2019-06-18 | 广东和信健康科技有限公司 | A kind of swin flu, second stream and adenovirus multiple nucleic acid detection primer probe groups and its kit |
| CN110273026A (en) * | 2019-06-20 | 2019-09-24 | 中山大学达安基因股份有限公司 | Respiratory tract infection Multiple detection kit and detection method |
| CN110331232A (en) * | 2019-06-20 | 2019-10-15 | 中山大学达安基因股份有限公司 | A kind of respiratory pathogen multiple nucleic acid detection kit |
| CN110358865A (en) * | 2019-07-25 | 2019-10-22 | 廊坊诺道中科医学检验实验室有限公司 | For detecting kit and its application of Respirovirus |
| CN110408725A (en) * | 2019-06-20 | 2019-11-05 | 中山大学达安基因股份有限公司 | Kit for respiratory pathogen Multiple detection |
| CN110468234A (en) * | 2019-08-09 | 2019-11-19 | 厦门安普利生物工程有限公司 | Multiple fluorescence quantitative PCR kit for 19 kinds of human respiratory viral diagnosis |
| CN110607379A (en) * | 2019-07-25 | 2019-12-24 | 廊坊诺道中科医学检验实验室有限公司 | Multiplex PCR detection method for simultaneously detecting multiple bacteria and mycoplasma and application thereof |
| CN111172324A (en) * | 2020-02-17 | 2020-05-19 | 深圳麦科田生物医疗技术有限公司 | Probe, primer, kit and detection method for detecting viruses A and B based on double loop-mediated isothermal amplification technology |
| CN111349720A (en) * | 2020-03-19 | 2020-06-30 | 中国医学科学院病原生物学研究所 | Nucleic acid reagent, kit, system and method for detecting respiratory tract infection virus |
| CN111549183A (en) * | 2020-06-08 | 2020-08-18 | 国药集团武汉血液制品有限公司 | Method for simultaneously detecting SARS-CoV-2 virus and other pathogens in plasma of convalescent coronary pneumonia convalescent patient |
| CN111808997A (en) * | 2020-08-27 | 2020-10-23 | 圣湘生物科技股份有限公司 | Composition, kit, application and method for detecting and typing 10 respiratory tract-related viruses |
| CN113249450A (en) * | 2021-06-29 | 2021-08-13 | 天津国际生物医药联合研究院 | Real-time fluorescent quantitative PCR method for detecting group A streptococcus nucleic acid |
| CN114058742A (en) * | 2022-01-18 | 2022-02-18 | 广州科方生物技术股份有限公司 | Primer probe composition, kit containing same and detection method thereof |
| CN114540544A (en) * | 2021-12-24 | 2022-05-27 | 廊坊诺道中科医学检验实验室有限公司 | Primer-probe combination for detecting respiratory viruses, kit and application thereof |
| CN116334265A (en) * | 2023-05-23 | 2023-06-27 | 爱科睿特生物医疗科技(南京)有限公司 | Multiplex RT-PCR method for simultaneously detecting four human respiratory tract infection bacteria, kit and application |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111321251B (en) * | 2020-04-16 | 2020-08-14 | 圣湘生物科技股份有限公司 | Composition, kit, method and application for detecting and typing pathogens causing respiratory tract infection |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060003352A1 (en) * | 2004-04-29 | 2006-01-05 | Lipkin W I | Mass tag PCR for mutliplex diagnostics |
| CN102212618A (en) * | 2011-05-22 | 2011-10-12 | 浙江大学 | Fourfold fluorescence quantitative PCR (Polymerase Chain Reaction) kit and application |
| CN102851391A (en) * | 2012-07-20 | 2013-01-02 | 宁波检验检疫科学技术研究院 | Human enterovirus four-color fluorescence RT-PCR detection kit and detection method |
| CN103993102A (en) * | 2014-03-07 | 2014-08-20 | 崔淑娟 | Multiple fluorescent PCR method and kit for simultaneous detection of human adenovirus, human mycoplasma pneumonia and bocavirus |
| CN104561377A (en) * | 2014-12-24 | 2015-04-29 | 华美生物工程有限公司 | Real-time fluorescent multiplex PCR (polymerase chain reaction) based kit for rapidly detecting common respiratory pathogens |
| CN105463129A (en) * | 2015-12-09 | 2016-04-06 | 深圳国际旅行卫生保健中心 | Double-fluorescent PCR detection primer, probe, reaction liquid and kit capable of detecting pathogens of respiratory tract |
-
2017
- 2017-03-30 CN CN201710201003.5A patent/CN107058622B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060003352A1 (en) * | 2004-04-29 | 2006-01-05 | Lipkin W I | Mass tag PCR for mutliplex diagnostics |
| WO2006073436A2 (en) * | 2004-04-29 | 2006-07-13 | The Trustees Of Columbia University In The City Of New York | Mass tag pcr for multiplex diagnostics |
| CN102212618A (en) * | 2011-05-22 | 2011-10-12 | 浙江大学 | Fourfold fluorescence quantitative PCR (Polymerase Chain Reaction) kit and application |
| CN102851391A (en) * | 2012-07-20 | 2013-01-02 | 宁波检验检疫科学技术研究院 | Human enterovirus four-color fluorescence RT-PCR detection kit and detection method |
| CN103993102A (en) * | 2014-03-07 | 2014-08-20 | 崔淑娟 | Multiple fluorescent PCR method and kit for simultaneous detection of human adenovirus, human mycoplasma pneumonia and bocavirus |
| CN104561377A (en) * | 2014-12-24 | 2015-04-29 | 华美生物工程有限公司 | Real-time fluorescent multiplex PCR (polymerase chain reaction) based kit for rapidly detecting common respiratory pathogens |
| CN105463129A (en) * | 2015-12-09 | 2016-04-06 | 深圳国际旅行卫生保健中心 | Double-fluorescent PCR detection primer, probe, reaction liquid and kit capable of detecting pathogens of respiratory tract |
Cited By (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107630098B (en) * | 2017-11-13 | 2019-11-08 | 湖南圣湘生物科技有限公司 | Fluorescent PCR detection architecture, kit and detection method for joint-detection various respiratory road bacterium |
| CN107630098A (en) * | 2017-11-13 | 2018-01-26 | 湖南圣湘生物科技有限公司 | Fluorescent PCR detection architecture, kit and detection method for joint-detection various respiratory road bacterium |
| CN107828633A (en) * | 2017-12-14 | 2018-03-23 | 苏州天隆生物科技有限公司 | A kind of nucleic acid extraction and reagent device, reagent and the method for detection |
| CN107828633B (en) * | 2017-12-14 | 2021-04-27 | 苏州天隆生物科技有限公司 | Reagent device, reagent and method for extracting and detecting nucleic acid |
| CN108374059A (en) * | 2018-03-01 | 2018-08-07 | 绍兴迅敏康生物科技有限公司 | Respiratory Syncytial Virus(RSV) kit for detecting nucleic acid and its detection method |
| CN109609692A (en) * | 2018-12-25 | 2019-04-12 | 北京卓诚惠生生物科技股份有限公司 | For detecting the nucleic acid reagent, kit, system and method for respiratory tract infection pathogen |
| CN109897917A (en) * | 2019-04-01 | 2019-06-18 | 广东和信健康科技有限公司 | A kind of swin flu, second stream and adenovirus multiple nucleic acid detection primer probe groups and its kit |
| CN110408725A (en) * | 2019-06-20 | 2019-11-05 | 中山大学达安基因股份有限公司 | Kit for respiratory pathogen Multiple detection |
| CN110331232A (en) * | 2019-06-20 | 2019-10-15 | 中山大学达安基因股份有限公司 | A kind of respiratory pathogen multiple nucleic acid detection kit |
| CN110273026A (en) * | 2019-06-20 | 2019-09-24 | 中山大学达安基因股份有限公司 | Respiratory tract infection Multiple detection kit and detection method |
| CN110331232B (en) * | 2019-06-20 | 2022-07-05 | 广州达安基因股份有限公司 | Multiple nucleic acid detection kit for respiratory pathogens |
| CN110408725B (en) * | 2019-06-20 | 2022-07-05 | 广州达安基因股份有限公司 | Kit for multiple detection of respiratory pathogens |
| CN110273026B (en) * | 2019-06-20 | 2022-07-05 | 广州达安基因股份有限公司 | Multiple detection kit and detection method for respiratory tract infection |
| CN110358865A (en) * | 2019-07-25 | 2019-10-22 | 廊坊诺道中科医学检验实验室有限公司 | For detecting kit and its application of Respirovirus |
| CN110607379A (en) * | 2019-07-25 | 2019-12-24 | 廊坊诺道中科医学检验实验室有限公司 | Multiplex PCR detection method for simultaneously detecting multiple bacteria and mycoplasma and application thereof |
| CN110468234A (en) * | 2019-08-09 | 2019-11-19 | 厦门安普利生物工程有限公司 | Multiple fluorescence quantitative PCR kit for 19 kinds of human respiratory viral diagnosis |
| CN111172324A (en) * | 2020-02-17 | 2020-05-19 | 深圳麦科田生物医疗技术有限公司 | Probe, primer, kit and detection method for detecting viruses A and B based on double loop-mediated isothermal amplification technology |
| CN111349720A (en) * | 2020-03-19 | 2020-06-30 | 中国医学科学院病原生物学研究所 | Nucleic acid reagent, kit, system and method for detecting respiratory tract infection virus |
| CN111549183A (en) * | 2020-06-08 | 2020-08-18 | 国药集团武汉血液制品有限公司 | Method for simultaneously detecting SARS-CoV-2 virus and other pathogens in plasma of convalescent coronary pneumonia convalescent patient |
| CN111808997A (en) * | 2020-08-27 | 2020-10-23 | 圣湘生物科技股份有限公司 | Composition, kit, application and method for detecting and typing 10 respiratory tract-related viruses |
| CN113249450A (en) * | 2021-06-29 | 2021-08-13 | 天津国际生物医药联合研究院 | Real-time fluorescent quantitative PCR method for detecting group A streptococcus nucleic acid |
| CN114540544A (en) * | 2021-12-24 | 2022-05-27 | 廊坊诺道中科医学检验实验室有限公司 | Primer-probe combination for detecting respiratory viruses, kit and application thereof |
| CN114058742A (en) * | 2022-01-18 | 2022-02-18 | 广州科方生物技术股份有限公司 | Primer probe composition, kit containing same and detection method thereof |
| CN114058742B (en) * | 2022-01-18 | 2022-04-22 | 广州科方生物技术股份有限公司 | Primer probe composition, kit containing same and detection method thereof |
| CN116334265A (en) * | 2023-05-23 | 2023-06-27 | 爱科睿特生物医疗科技(南京)有限公司 | Multiplex RT-PCR method for simultaneously detecting four human respiratory tract infection bacteria, kit and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107058622B (en) | 2020-03-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107058622A (en) | A kind of kit of multiple fluorescence PCR method joint-detection respiratory pathogen | |
| WO2021174674A1 (en) | Composition, kit and method for detecting and typing coronaviruses | |
| US20230287524A1 (en) | Composition, kit and method for detecting and classifying pathogens causing respiratory tract infection, and application | |
| CN105018648B (en) | A kind of kit and its application for being used to detect Respirovirus | |
| CN110468234A (en) | Multiple fluorescence quantitative PCR kit for 19 kinds of human respiratory viral diagnosis | |
| CN105112559B (en) | A kind of kit and its application for being used to detect coronavirus | |
| CN103060451B (en) | A kind of mycoplasma pneumoniae MP detection kit | |
| CN102337351B (en) | Typing detection kit for influenza virus | |
| CN105039597B (en) | A kind of kit and its application for being used to detect influenza virus | |
| CN109517927A (en) | A kind of A type, influenza B virus rapid typing detection reagent box and its application | |
| CN112359145B (en) | Multiple primers and kit for rapidly detecting influenza A, influenza B and novel coronavirus | |
| CN1746319A (en) | Detection of real-time fluorescent polyase chain reaction of respiratory pathogen | |
| WO2023087868A1 (en) | Compositions, kits, methods for detecting and identifying pathogens that cause respiratory tract infections and use thereof | |
| CN111635950A (en) | Kit and method for rapidly detecting bordetella pertussis | |
| CN104450970A (en) | Kit and method for identifying porcine reproductive and respiratory syndrome viruses | |
| CN106520923A (en) | Kit and method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof | |
| CN105018488B (en) | Kit and its application for detecting Respirovirus | |
| CN102776283A (en) | Visualized loop-mediated isothermal amplification kit for detecting Haemophilus parasuis | |
| CN111424114A (en) | 2019-nCoV novel coronavirus saliva detection biomarker and application thereof | |
| CN110157823A (en) | Primed probe group, kit and its application of RAA Fluorometric assay Yersinia pestis | |
| CN108504774B (en) | Method for detecting rhinovirus typing and enterovirus | |
| CN101575649B (en) | Quick detection technology for H9 type avian influenza virus | |
| CN108559790A (en) | The kit and its application method of three kinds of respiratory pathogens are detected based on micro-fluidic chip | |
| WO2017092483A1 (en) | Kit for diagnosing tuberculosis through detecting free nucleic acid and use thereof | |
| WO2023109031A1 (en) | Respiratory pathogen detection kit, and preparation method therefor and use thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |