CN107815491A - A kind of kit and its application method of quick single-minded detection Mycobacterium tuberculosis - Google Patents
A kind of kit and its application method of quick single-minded detection Mycobacterium tuberculosis Download PDFInfo
- Publication number
- CN107815491A CN107815491A CN201711167560.6A CN201711167560A CN107815491A CN 107815491 A CN107815491 A CN 107815491A CN 201711167560 A CN201711167560 A CN 201711167560A CN 107815491 A CN107815491 A CN 107815491A
- Authority
- CN
- China
- Prior art keywords
- sequence
- seq
- mycobacterium tuberculosis
- kit
- reaction
- 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
- C12Q1/68—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
-
- 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
Abstract
The present invention relates to a kind of kit and its application method of quick single-minded detection Mycobacterium tuberculosis, the kit includes working standard, isothermal amplification reactions liquid and negative controls, working standard includes the positive control plasmid of the special conserved sequence of Mycobacterium tuberculosis, and special conserved sequence is SEQ ID No.1；Isothermal amplification reactions liquid includes 3 pairs of primers：Outer primer F3, its sequence are SEQ ID No.2；Outer primer B3, its sequence are SEQ ID No.3；Inner primer FIP, its sequence are SEQ ID No.4；Inner primer BIP, its sequence are SEQ ID No.5；Ring primer LB, its sequence are SEQ ID No.6；Ring primer LF, its sequence are SEQ ID No.7；Negative controls are the sputum DNA extract solutions of normal person.The kit quickly single-minded can detect Mycobacterium tuberculosis, and sensitivity and specificity are very good.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of kit of quick single-minded detection Mycobacterium tuberculosis and
Its application method.
Background technology
Mycobacterium tuberculosis (Mycobacterium tuberculosis) is to cause pathogen lungy, can be invaded complete
Each organ of body, but using pulmonary tuberculosis to be most common.Tuberculosis is still important infectious disease so far.According to WHO, 800 are there are about every year
Ten thousand new cases occur, and at least 3,000,000 people die from the disease.The death rate reaches 200-300 people/100,000 before the founding of the state in China, occupies various diseases
First of the sick cause of death, living standards of the people improve after the founding of the state, and hygienic state improves, and has particularly carried out mass prevention and mass treatment, children
Universal bcg vaccination, morbidity and mortality lungy are greatly lowered.It should be noted that some areas are because of AIDS in the world
The reasons such as disease, drug abuse, the application of immunodepressant, excessive drinking and poverty, the incidence of disease are again on the rise.Therefore, to tuberculosis branch
Timely, efficient, the specific detection of bacillus causes the effective means of illness as prevention, treatment mycobacterium tuberculosis.
Bacteria Culture and plate coating checking are one of etiological diagnosis technologies being most widely used, and wherein sputum culture accounts for
More than 50%, but because the complexity of the sputum specimen lack of standardization and microorganism of collection and the correlation of human body causes traditional habit journey
Its clinical coincidence rate of the sputum sample bacteriological analysis of sequence is not high.
Antibody test, detection of nucleic acids and drug sensitive culture medium inspection are based primarily upon to the detection reagent of mycobacterium tuberculosis both at home and abroad
Survey etc..At present, detection of nucleic acids has turned into the main method of mycobacterium tuberculosis detection；To the detection of nucleic acids master of mycobacterium tuberculosis
Have with Types Below：PCR- fluorescence probe methods, fluorescent PCR melting curve method, PCR- revert dot blot hybridizations, DNA microarray chip
Method etc..But because antibody test sensitivity is relatively low, false positive phenomenon is more, detection of nucleic acids needs the large-scale instruments such as fluorescent PCR instrument,
And need to take the features such as longer to pathogen progress nucleic acid extraction, follow-up fluoroscopic examination in advance.
The isothermal amplification technology (loop-mediated isothermal amplification, LAMP) of ring mediation is one
The new nucleic acid amplification technologies of kind, it is designed using the archaeal dna polymerase with strand-displacement activity, and for DNA 6 isolated areas
Two pairs or the special primer of three teams, under 60-65 DEG C of reaction condition 20-30 minutes can complete to react.LAMP has quick
The characteristics of simple and high specificity, it can be expected to substitute detection of the traditional PCR method to microorganism.Therefore, we are for knot
One section of specific primer sequences of core mycobacterium, have invented a kind of kit, and it being capable of quick special detection M tuberculosis bar
Bacterium.
The content of the invention
The present invention provides a kind of kit and its application method of quick single-minded detection Mycobacterium tuberculosis, and it can be real-time
To in patient's sputum and swab equal samples Mycobacterium tuberculosis carry out fast accurate detection.
The technical scheme that the present invention solves above-mentioned technical problem is as follows：A kind of examination of quick single-minded detection Mycobacterium tuberculosis
Agent box, it includes working standard, isothermal amplification reactions liquid and negative controls,
The working standard includes the positive control plasmid of the special conserved sequence of Mycobacterium tuberculosis, the special guarantor
It is SEQ ID No.1 to keep sequence；
The isothermal amplification reactions liquid includes 3 pairs of primers：Outer primer F3, its sequence are SEQ ID No.2；Outer primer
B3, its sequence are SEQ ID No.3；Inner primer FIP, its sequence are SEQ ID No.4；Inner primer BIP, its sequence are SEQ ID
No.5；Ring primer LB, its sequence are SEQ ID No.6；Ring primer LF, its sequence are SEQ ID No.7；
The negative controls are the sputum DNA extract solutions of normal person.
On the basis of above-mentioned technical proposal, the present invention can also have following further specifically chosen.
It is cloned into specifically, the positive control plasmid synthesizes fragment corresponding to special conserved sequence in pGEM-T carriers
Built-up, its concentration is 104copies/μL。
Specifically, the solvent of the isothermal amplification reactions liquid is distilled water, the isothermal amplification reactions liquid contains following dense
Each solute of degree：8U/20 μ L Bst archaeal dna polymerases；8mM MgSO4；2.4mM each dNTPs；20mM KCl；20mM
(NH4)2SO4；40Mm Tris；40Mm HCl；0.2wt%TritonX-100；5pmol/20 μ L outer primers F3；5pmol/20μL
Outer primer B3；40pmol inner primers FIP；40pmol inner primers BIP；20pmol ring primers LB；20pmol ring primers LB；1.25×
EvaGreen fluorescent dyes.
In addition, present invention also offers a kind of side of the kit using above-mentioned quick single-minded detection Mycobacterium tuberculosis
Method, it comprises the following steps：
S1. sputum DNA preparation of specimen to be measured；
S2. three by-reaction pipes are taken, each reaction tube adds isothermal amplification reactions liquid described in 20 μ L, then in the first reaction
5 μ L sputum DNA samples to be measured are added in pipe, 5 μ L working standards is added in the second reaction tube, is added in the 3rd reaction tube
5 μ L negative controls, then three reaction tubes are placed under constant temperature and carry out amplified reaction；
S3. after the completion of amplified reaction, with ELIASA etc. read in three anti-pipes of reaction the fluorescent value of reaction solution and compare with
Determine whether contain Mycobacterium tuberculosis in sputum DNA samples to be measured.The real time measure fluorescent value during amplified reaction, according to
Amplification curve is to determine whether contain Mycobacterium tuberculosis in sputum DNA samples to be measured.
Specifically, the reaction temperature of amplified reaction is 65 DEG C, reaction time 60min.
Compared with prior art, the beneficial effects of the invention are as follows：
The present invention has filtered out one section of spy for including 300bp by the way that Mycobacterium tuberculosis gene order is compared
Different conserved sequence, then special conserved sequence devises 3 pairs of special primers for more than, and 3 pairs of special primers gather with Bst DNA
Synthase, EvaGreen fluorescent dyes (observe by the naked eye turbidity can be detection band carry out error, the sensitivity of detection is not also high, therefore
With the addition of fluorescent dye), necessary inorganic salts and buffer solution constitute isothermal amplification reactions liquid, the DNA of Mycobacterium tuberculosis exists
In above-mentioned isothermal amplification reactions liquid can rapid amplifying, monitored in real time by fluorescence, so as to realize to the quick of mycobacterium tuberculosis
Detection, can be applied to the quick detection of mycobacterium tuberculosis in sputum or Nasopharyngeal swabs sample, easy to operate, detection is convenient；With
Above-mentioned isothermal amplification reactions liquid, the working standard containing the special conserved sequence of Mycobacterium tuberculosis and negative controls constitute
The kit of quick single-minded detection Mycobacterium tuberculosis, it is each with testing sample DNA, working standard and negative controls during use
5 μ L, it is separately added into three reaction tubes equipped with isothermal amplification reactions liquid described in 20 μ L, during amplified reaction, to prison in real time
The fluorescent value measured is compared, you can is rapidly completed detection.
The sensitivity of kit provided by the invention and specificity are very good：Coordinate with fluorescence detection device, its is minimum
Detect to be limited to 10copies/ μ L, to common lung pathogenic bacteria pseudomonas aeruginosa, streptococcus pneumonia, haemophilus influenzae, thermophilic
Detected after the extraction such as lung Legionella, mycoplasma pneumoniae, Klebsiella Pneumoniae, staphylococcus aureus DNA, specific test
Only have mycobacterium tuberculosis to have amplification curve in experiment, other common lung pathogenic germs are without amplification curve.Reagent of the present invention
Box is adapted to detection scene and basic medical unit popularization and application because of quick, Gao Min, simple to operate, low cost and other advantages.
Brief description of the drawings
Fig. 1 is the amplification curve of the specific test carried out for kit provided by the invention；
Fig. 2 is the result of the sensitivity test carried out for kit provided by the invention；
Fig. 3 is the amplification curve that the clinical sample carried out for kit provided by the invention is tested.
Embodiment
Technical scheme is described in further detail below in conjunction with drawings and the specific embodiments, example
It is served only for explaining the present invention, is not intended to limit the scope of the present invention.
A kind of kit of quick single-minded detection Mycobacterium tuberculosis, it includes working standard, isothermal amplification reactions liquid
And negative controls, the working standard includes the positive control plasmid of the special conserved sequence of Mycobacterium tuberculosis, described
Special conserved sequence is SEQ ID No.1；The isothermal amplification reactions liquid includes 3 pairs of primers：Outer primer F3, its sequence are
SEQ ID No.2；Outer primer B3, its sequence are SEQ ID No.3；Inner primer FIP, its sequence are SEQ ID No.4；Inner primer
BIP, its sequence are SEQ ID No.5；Ring primer LB, its sequence are SEQ ID No.6；Ring primer LF, its sequence are SEQ ID
No.7；The negative controls are the sputum DNA extract solutions of normal person.
1. the compound method of working standard is specific as follows：
(1) the mycobacterium tuberculosis conserved sequence (SEQ ID No.1) of one section of 300bp length is synthesized with chemical synthesis.
(2) synthesis fragment is cloned into pGEM-T carriers, builds mycobacterium tuberculosis positive control plasmid.
(3) obtained positive control plasmid is transformed into bacillus coli DH 5 alpha, and trained with the LB containing 1% ampicillin
After supporting 37 DEG C of mass propgations of base, carried with the big extraction reagent kit of high pure plasmid (DP116) of TIANGEN Biotech (Beijing) Co., Ltd.
Take plasmid.
(4) plasmid is purified and quantified, be diluted to 104Copies/mL, produce working standard.
2. the concrete composition of isothermal amplification reactions liquid is as follows：
Solvent is distilled water, and its interior solute is：8U/20 μ L Bst archaeal dna polymerases (are bought from NEW ENGLAND
BioLabs, article No.：M0275)、8mM MgSO4、2.4mM each dNTPs、40mM Tris-HCl、20mM KCl、20mM
(NH4)2SO4, 0.2%TritonX-100,5pmol/20 μ L outer primers F3；5pmol/20 μ L outer primers B3；40pmol inner primers
FIP；40pmol inner primers BIP；20pmol ring primers LB；20pmol ring primers LB；1.25 × EvaGreen fluorescent dyes
(Biotium, article No. 31000).
3. negative controls are the DNA extract solutions of normal person's sputum, extracting method is as follows：
A. the sputum of 50 μ L normal persons is taken in the centrifuge tube of sterilizing.
B. 50 μ L Lysis Buffer solution are added, are well mixed.
C.80 DEG C thermal denaturation is after 15 minutes, low-speed centrifugal, then takes supernatant in the centrifuge tube of new sterilizing, produces normal
The DNA extract solutions (experiment is immediately available for after extraction or -20 DEG C freeze) of people's sputum.
4. sputum DNA preparation of specimen to be measured
Using the sputum of possible Mycobacterium tuberculosis carrier as sample extraction, sample to be tested preparation method with it is above-mentioned just
Ordinary person's sputum DNA extraction liquor manufacture methods are identical.
5. reaction system is prepared and amplified reaction
At least three reaction tubes：Add 20 μ L isothermal amplification reactions liquids and 5 μ L sputum DNA samples to be measured in first reaction tube；
Add 20 μ L isothermal amplification reactions liquids and 5 μ L working standards in second reaction tube；In 3rd reaction tube plus 20 μ L constant-temperature amplifications are anti-
Answer liquid and 5 μ L negative controls.
Amplified reaction：Above-mentioned reaction tube is reacted into 60min, real-time testing fluorescent value, according to amplification in the environment of 65 DEG C
Curve judges whether carry Mycobacterium tuberculosis in sputum DNA samples to be measured, is judged to carrying if having amplification curve.
6. detection
A. specific test
To common lung pathogenic bacteria pseudomonas aeruginosa, streptococcus pneumonia, haemophilus influenzae, legionella pneumophilia, pneumonia
Detected after the extraction such as mycoplasma, Klebsiella Pneumoniae, staphylococcus aureus DNA with the kit of the present invention.As a result such as
Shown in Fig. 1, only Mycobacterium tuberculosis has amplification curve in specific test experiment, and other common lung pathogenic germs are without expansion
Increase curve, show kit specificity provided by the invention preferably.
B. sensitivity experiment
It is 10 by concentration4Copies/ μ l Mycobacterium tuberculosis standard items (i.e. working standard) press 10 respectively3、102、
10copies/ μ l gradient dilutions, afterwards respectively as sample to be tested DNA, it is detected using the kit of the present invention.Knot
Fruit can detect that about 10copies/ μ l are copied as shown in fig. 2, it can be seen that the kit is minimum, have very high sensitive
Degree.
C. the detection of the clinical doubtful bacterial strain of Mycobacterium tuberculosis
Collect BJ Chest Science Hospital and identified 6 sputums containing mycobacterium tuberculosis clinical case, according to sputum to be measured
The method extraction DNA of DNA preparation of specimen, is then detected with the kit of the present invention.As a result as shown in figure 3, to 6 tuberculosis
Mycobacteria clinical case is detected, wherein 6 are that Mycobacterium tuberculosis is positive, comprehensive positive rate is 100%.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc., it should be included in the scope of the protection.
Sequence table
<110>Fu Luolang（Beijing）Bio tech ltd
<120>A kind of quick single-minded detection mycobacterium tuberculosis kit and its application method
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 300
<212> DNA
<213>Mycobacterium tuberculosis (Mycobacterium tuberculosis)
<400> 1
ccgatcgccc catcgaccta ctacgaccac atcaaccggg agcccagccg ccgcgagctg 60
cgcgatggcg aactcaagga gcacatcagc cgcgtccacg ccgccaacta cggtgtttac 120
ggtgcccgca aagtgtggct aaccctgaac cgtgagggca tcgaggtggc cagatgcacc 180
gtcgaacggc tgatgaccaa actcggcctg tccgggacca cccgcggcaa agcccgcagg 240
accacgatcg ctgatccggc cacagcccgt cccgccgatc tcgtccagcg ccgcttcgga 300
<210> 2
<211> 16
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 2
tcaaccggga gcccag 16
<210> 3
<211> 16
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 3
tttgccgcgg gtggtc 16
<210> 4
<211> 43
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 4
cgtaaacacc gtagttggcg gcttttgcgc gatggcgaac tca 43
<210> 5
<211> 46
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 5
gtgcccgcaa agtgtggcta acttttagtt tggtcatcag ccgttc 46
<210> 6
<211> 16
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 6
acgcggctga tgtgct 16
<210> 7
<211> 17
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 7
aggtggccag atgcacc 17
Claims (5)
1. a kind of kit of quick single-minded detection Mycobacterium tuberculosis, it is characterised in that including working standard, constant-temperature amplification
Reaction solution and negative controls,
The working standard is the positive control plasmid for including the special conserved sequence of Mycobacterium tuberculosis, the specifically conservative sequence
It is classified as SEQ ID No.1；
The isothermal amplification reactions liquid includes 3 pairs of primers：Outer primer F3, its sequence are SEQ ID No.2；Outer primer B3, its
Sequence is SEQ ID No.3；Inner primer FIP, its sequence are SEQ ID No.4；Inner primer BIP, its sequence are SEQ ID
No.5；Ring primer LB, its sequence are SEQ ID No.6；Ring primer LF, its sequence are SEQ ID No.7；
The negative controls are the sputum DNA extract solutions of normal person.
2. the kit of a kind of quick single-minded detection Mycobacterium tuberculosis according to claim 1, it is characterised in that described
Positive control plasmid synthesizes fragment corresponding to special conserved sequence and is cloned into built-up in pGEM-T carriers, and its concentration is
104copies/μL。
3. the kit of a kind of quick single-minded detection Mycobacterium tuberculosis according to claim 1, it is characterised in that described
The solvent of isothermal amplification reactions liquid is distilled water, and the isothermal amplification reactions liquid contains each solute of following concentration：8U/20μL
Bst archaeal dna polymerases；8mM MgSO4；2.4mM each dNTPs；20mM KCl；20mM(NH4)2SO4；40Mm Tris-HCl；
0.2wt%TritonX-100；5pmol/20 μ L outer primers F3；5pmol/20 μ L outer primers B3；40pmol inner primers FIP；
40pmol inner primers BIP；20pmol ring primers LB；20pmol ring primers LB；1.25 × EvaGreen fluorescent dyes.
A kind of 4. side of the kit of quick single-minded detection Mycobacterium tuberculosis using as described in any one of claims 1 to 3
Method, it is characterised in that comprise the following steps：
S1. sputum DNA preparation of specimen to be measured；
S2. three by-reaction pipes are taken, each reaction tube adds isothermal amplification reactions liquid described in 20 μ L, then in the first reaction tube
5 μ L sputum DNA samples to be measured are added, 5 μ L working standards is added in the second reaction tube, 5 μ L is added in the 3rd reaction tube
Negative controls, then three reaction tubes are placed under constant temperature and carry out amplified reaction；
S3. the real time measure three is reacted the fluorescent value of reaction solution in anti-pipes and compared to determine phlegm to be measured during amplified reaction
Whether contain Mycobacterium tuberculosis in liquid DNA samples.
5. a kind of method of kit using quick single-minded detection Mycobacterium tuberculosis according to claim 4, it is special
Sign is that the reaction temperature of amplified reaction is 65 DEG C, reaction time 60min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711167560.6A CN107815491B (en) | 2017-11-21 | 2017-11-21 | Kit for rapidly and specifically detecting mycobacterium tuberculosis and use method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711167560.6A CN107815491B (en) | 2017-11-21 | 2017-11-21 | Kit for rapidly and specifically detecting mycobacterium tuberculosis and use method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107815491A true CN107815491A (en) | 2018-03-20 |
| CN107815491B CN107815491B (en) | 2020-12-29 |
Family
ID=61610100
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711167560.6A Active CN107815491B (en) | 2017-11-21 | 2017-11-21 | Kit for rapidly and specifically detecting mycobacterium tuberculosis and use method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107815491B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101157954A (en) * | 2007-09-13 | 2008-04-09 | 中华人民共和国徐州出入境检验检疫局 | Concretion mycobacterium nucleic acid screening method based on loop-mediated isothermal amplification technique |
| CN101302553A (en) * | 2008-05-30 | 2008-11-12 | 广州华峰生物科技有限公司 | Mycobacterium tuberculosis gene rapid diagnosis reagent kit base on loop-mediated isothermal amplification technology and detection method thereof |
| CN101575640A (en) * | 2009-03-12 | 2009-11-11 | 华南农业大学 | Primer group for bovine tuberculosis mycobacterium detection, rapid detection method and detection kit |
| CN102399901A (en) * | 2011-12-21 | 2012-04-04 | 中国人民解放军疾病预防控制所 | Loop-mediated isothermal amplification (LAMP) detection method for mycobacterium tuberculosis, and special primer and kit thereof |
| CN102888455A (en) * | 2012-09-14 | 2013-01-23 | 珠海市银科医学工程有限公司 | Kit for detecting mycobacterium tuberculosis based on loop-mediated isothermal amplification, and primers |
| CN103667425A (en) * | 2012-08-30 | 2014-03-26 | 益生生技开发股份有限公司 | Method and kit for detecting tubercle bacillus |
| CN103937884A (en) * | 2014-03-28 | 2014-07-23 | 中国检验检疫科学研究院 | Loop-mediated isothermal amplification kit for detecting mycobacterium tuberculosis and application method of kit |
| CN105907861A (en) * | 2016-05-05 | 2016-08-31 | 广州金域医学检验中心有限公司 | Method and primer for quick detection and classification of mycobacteria |
| CN107058517A (en) * | 2017-03-13 | 2017-08-18 | 新乡医学院第附属医院 | A kind of kit and detection method that detection is infected for mycobacterium tuberculosis |
-
2017
- 2017-11-21 CN CN201711167560.6A patent/CN107815491B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101157954A (en) * | 2007-09-13 | 2008-04-09 | 中华人民共和国徐州出入境检验检疫局 | Concretion mycobacterium nucleic acid screening method based on loop-mediated isothermal amplification technique |
| CN101302553A (en) * | 2008-05-30 | 2008-11-12 | 广州华峰生物科技有限公司 | Mycobacterium tuberculosis gene rapid diagnosis reagent kit base on loop-mediated isothermal amplification technology and detection method thereof |
| CN101575640A (en) * | 2009-03-12 | 2009-11-11 | 华南农业大学 | Primer group for bovine tuberculosis mycobacterium detection, rapid detection method and detection kit |
| CN102399901A (en) * | 2011-12-21 | 2012-04-04 | 中国人民解放军疾病预防控制所 | Loop-mediated isothermal amplification (LAMP) detection method for mycobacterium tuberculosis, and special primer and kit thereof |
| CN103667425A (en) * | 2012-08-30 | 2014-03-26 | 益生生技开发股份有限公司 | Method and kit for detecting tubercle bacillus |
| CN102888455A (en) * | 2012-09-14 | 2013-01-23 | 珠海市银科医学工程有限公司 | Kit for detecting mycobacterium tuberculosis based on loop-mediated isothermal amplification, and primers |
| CN103937884A (en) * | 2014-03-28 | 2014-07-23 | 中国检验检疫科学研究院 | Loop-mediated isothermal amplification kit for detecting mycobacterium tuberculosis and application method of kit |
| CN105907861A (en) * | 2016-05-05 | 2016-08-31 | 广州金域医学检验中心有限公司 | Method and primer for quick detection and classification of mycobacteria |
| CN107058517A (en) * | 2017-03-13 | 2017-08-18 | 新乡医学院第附属医院 | A kind of kit and detection method that detection is infected for mycobacterium tuberculosis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107815491B (en) | 2020-12-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Fenollar et al. | Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria | |
| Mühl et al. | Activity and DNA contamination of commercial polymerase chain reaction reagents for the universal 16S rDNA real-time polymerase chain reaction detection of bacterial pathogens in blood | |
| US8288524B2 (en) | Molecular diagnosis of bacteremia | |
| Båverud et al. | Real-time PCR for detection and differentiation of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus | |
| Maďarová et al. | A real-time PCR diagnostic method for detection of Naegleria fowleri | |
| Albay et al. | The evaluation of FASTPlaqueTBTM test for the rapid diagnosis of tuberculosis | |
| Miyashita et al. | Multiplex PCR for the simultaneous detection of Chlamydia pneumoniae, Mycoplasma pneumoniae and Legionella pneumophila in community-acquired pneumonia | |
| CN112831580B (en) | Reaction system for detecting vibrio parahaemolyticus DNA, kit and application thereof | |
| Padmavathy et al. | Rapid and sensitive detection of major uropathogens in a single-pot multiplex PCR assay | |
| CN101429539B (en) | Reagent kit for real time fluorescence quantitative PCR detection of bacillus pyocyaneus | |
| Fan et al. | Development of a loop-mediated isothermal amplification assay for sensitive and rapid detection of Cronobacter sakazakii | |
| Zhang et al. | Development of a loop-mediated isothermal amplification assay for the detection of Mycobacterium bovis | |
| García-Quintanilla et al. | Simultaneous identification of Mycobacterium genus and Mycobacterium tuberculosis complex in clinical samples by 5′-exonuclease fluorogenic PCR | |
| Clancy et al. | Development of internally controlled duplex real-time NASBA diagnostics assays for the detection of microorganisms associated with bacterial meningitis | |
| Gupta et al. | Single-step PCR for detection of Brucella melitensis from tissue and blood of goats | |
| CN112301159A (en) | RDA method and kit for rapidly detecting influenza B virus | |
| JP4806749B2 (en) | Bordetella pertussis gene detection method using LAMP method and primer set used in this method | |
| Wilson et al. | Development of a simplified polymerase chain reactionenzyme immunoassay for the detection of Chlamydia pneumoniae | |
| CN102936625B (en) | Molecular motor biosensor kit used in vibrio parahaemolyticus molecular typing | |
| RU2542395C1 (en) | Set of reagents and method for detecting dna of causative agents of plague, anthrax and tularemia by pcr method with hybridisation-fluorescent recording of results | |
| CN107815491A (en) | A kind of kit and its application method of quick single-minded detection Mycobacterium tuberculosis | |
| CN107828910A (en) | A kind of kit and its application method of quick single-minded detection Bordetella pertussis | |
| CN108048586A (en) | A kind of detection method of ox kind Brucella sp attenuated vaccine strain S19 | |
| CN113584221A (en) | Kit for rapidly and specifically detecting influenza A virus and use method thereof | |
| CN107937584B (en) | Meat salmonella molecular detection kit and non-diagnostic detection method 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 |