CN108546768A - The sandwich DNA hybridization of staphylococcus aureus, which quickly detects, uses probe, kit and detection method - Google Patents
The sandwich DNA hybridization of staphylococcus aureus, which quickly detects, uses probe, kit and detection method Download PDFInfo
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- CN108546768A CN108546768A CN201810449424.4A CN201810449424A CN108546768A CN 108546768 A CN108546768 A CN 108546768A CN 201810449424 A CN201810449424 A CN 201810449424A CN 108546768 A CN108546768 A CN 108546768A
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- 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
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- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
Abstract
The invention discloses a kind of sandwich DNA hybridization of staphylococcus aureus quickly detection probe, kit and detection methods.The probe includes a capture probe and a detection probe, and kit includes one piece of 96 orifice plate for being coated with poly dT, pretreatment liquid, bacterium solution lysate, lysis buffer, nucleic acid hybridization solution, probe liquid, cleaning solution, developing solution, terminate liquid, positive control solution and negative controls.All processes can be completed in 2h using this kit, TMB developing solutions observe color change or multi-function microplate reader measures OD values and judges result by being added.The present invention can quickly, it is a large amount of, specifically detect target sequence, and it is easy to operate, do not need expensive instrument and reagent, can not there is no technical requirement, detection time short operating personnel with naked eyes judging result.
Description
Technical field
The invention belongs to biochip fields, and in particular to a kind of sandwich DNA hybridization technologies of SA are quickly detected with probe, examination
Agent and its detection method.
Background technology
Staphylococcus aureus (Staphylococcus aureus, SA) is a kind of common gram-positive bacteria, extensively
It is distributed in the surface of environment, animal and human body, it is difficult to be eliminated.The purulent inflammation for often causing skin, tissue, organ, may be used also
It grows and breeds in food such as dairy products, meat products, fish, tinned food, secrete intestines poison related with virulence and pathogenicity
Plain (staphylococcal enterotoxins, SETs), leads to food poisoning, can cause the sense of people and many animals
Dye, death.In addition to this, staphylococcus aureus its also with Kawasaki disease, toxic shock syndrome (toxic shock
Syndrome) closely related.It is even abused with lack of standardization use of antibiotic, many staphylococcus aureuses has occurred
Endurance strain, especially methicillin resistant strain (MRSA, Methicillin-Resistant S.aureus) are complete
World-wide prevalence.It is reported according to national drug-resistant bacteria monitoring net national Bacterial resistance surveillance in 2015,2012~2015,
MRSA recall rates are 35% or more, and the appearance of antibody-resistant bacterium is so that antibiotic is substantially reduced as the effect of medical procedure.
Currently, the detection method of staphylococcus aureus mainly have biochemical identification method, immunological method, PC R technologies and
Biochip technology etc..Conventional biochemical detection method is most widely used, as a result accurately and reliably, but operates time and effort consuming, is not suitable for
The inspection of batch samples；Enzyme-linked immunosorbent assay (ELISA) is unstable, has changeability and dimensional instability；PCR is vulnerable to
Sample type interferes, and influences detection sensitivity；Sensibility is high in the detection for biochip technology, high specificity, but complicated for operation,
Poor for low concentration bacterium detection result, false positive rate is also higher, and needs expensive chip point sample instrument and scanner.
It there is no the kit using sandwich DNA hybridization technology detection SA both at home and abroad at present.
Invention content
In order to overcome the deficiencies of the prior art, the first purpose of the invention is to provide a kind of staphylococcus aureus is sandwich
Quickly detection probe, second purpose are to provide the kit using the probe to DNA hybridization technology, and third purpose is to provide
The application method of the kit of above-mentioned detection probe.
In order to realize that above-mentioned first purpose present invention adopts the following technical scheme that：The sandwich DNA of staphylococcus aureus is miscellaneous
Quickly detection probe, including SA capture probes and SA detection probes is handed over, the wherein DNA sequence dna of SA capture probes is SEQ ID
The DNA sequence dna of NO.1, SA detection probe is SEQ ID NO.2.
In order to realize that a kind of sandwich DNA hybridization of staphylococcus aureus of above-mentioned second purpose present invention offer quickly detects
With kit, including be coated with 96 orifice plates of poly dT, pretreatment liquid, bacterium solution lysate, lysis buffer, nucleic acid hybridization solution,
Probe liquid, cleaning solution, developing solution, terminate liquid, positive control solution and negative controls, the dosage that one of micropore reacts are：
The probe liquid consists of the following compositions：
DNA sequence dna is the 16 μ L of SA capture probes of 6 μm of ol/L of SEQ ID NO.1；
DNA sequence dna is the 16 μ L of SA detection probes of 6 μm of ol/L of SEQ ID NO.2；
Total 32 μ L；
The pretreatment liquid, volume are 30 μ L；
Lysate is obtained by the bacterium solution lysate and lysis buffer cracking, and volume is 30 μ L (two bottles of dense bacterium
Liquid lysate is dry powder-shaped, and every bottle need to use 6mL lysis buffers to crack；The lysis buffer, volume 12mL)；
The nucleic acid hybridization solution, volume are 96 μ L；
The cleaning solution uses after needing 10 times of dilutions, and volume is 750 μ L；
The developing solution, volume are 150 μ L；
The terminate liquid, volume are 100 μ L.
Further, in mentioned reagent box, the positive control solution is SA bacterium solutions, volume 5mL；The negative control
Liquid is non-SA bacterium solutions, volume 5mL.
In order to realize the bright third purpose of this law, it is fast that the present invention has proposed a kind of sandwich DNA hybridization of staphylococcus aureus
Fast detection method, includes the following steps：
(1) increase bacterium and cultivate prepare liquid 48h with pancreas peptone soybean broth (TSB), obtain measuring samples liquid；
(2) bacterium solution cracking reaction by the ratio of measuring samples liquid, pretreatment liquid and lysate 4 ﹕, 1 ﹕ 1 by volume in glass
It is mixed in glass pipe, is put into 37 DEG C of water-baths and cultivates 5 minutes.
(3) bacterium solution that hybridization reaction is drawn after the cracking of 150 μ L steps (2) is added in the micropore for being coated with poly dT, then
By volume 2:1 ratio mixed nucleus acid hybridization liquid and probe liquid are drawn 125 μ L and are added in the micropore containing bacterium solution, at room temperature
It is incubated in being placed in incubator within 2 minutes with 150rpm speed rocker on rail mounted shaking table；With 1 × PBST after incubation
Buffer board-washings 5 times add and are incubated at room temperature 20 minutes in 150 μ L TMB developing solutions to each micropore, are eventually adding 100 μ L
The sulfuric acid terminate liquid of 2M is put into the absorbance read in multi-function microplate reader at 450nm.
(4) result judgement (bacterium solution OD values-blank control OD values)/(negative control OD value-blank control OD values) >=2.1
It can be determined as the positive, and become blue after developing solution addition micropore, the more deep bacteria concentration to be measured of color is higher.
The principle of the present invention is：According to staphylococcus aureus heat stable nuclease (nuc) genetic characteristics, separately design special
Property two kinds of probes of capture probe and detection probe, target nucleic acids segment is anchored by capture probe, detection probe end mark
Remember that horseradish peroxidase, two kinds of probes are combined with object bacteria target gene specific, it is ensured that the accuracy of testing result.As a result it examines
The measurement of OD values can be carried out, detection sensitivity can reach 55CFU/mL by being catalyzed color reaction by surveying.
Sandwich DNA hybridization is a kind of easy, quick, high degree of specificity hybridization.By sandwich DNA hybridization
Technology is compared with round pcr, it is possible to find the technology is equivalent to or excellent in the indexs such as sensitivity, specificity and detection range
In round pcr, false positive and false negative rate are low, as a result accurately, farthest reduce influence of the sample substrate to analysis result；
The Zengjing Granule time is short, and without extracting nucleic acid, sample treatment program is simple, the error for reducing test operation and its bringing, as a result
Intuitively.Existing SA detection cycles are longer, about 1~2 day, cumbersome, and the kit of the present invention only needs 2 hours.The present invention
Hybridization is applied to the quick detection of SA using sandwich DNA hybridization probe, specificity, accuracy are than common
PCR method higher, while expensive instrument input can be exempted, the popularization convenient for entering and leaving the border with food security detection of pathogens is answered
With.
It is an advantage of the invention that (1), sample pretreatment are simple, without extracting nucleic acid, directly using bacterium solution as masterplate；(2)、
High specific：It is combined with target pathogenic bacteria target gene to two probe specificities, whether is developed the color just according to horseradish peroxidase
It can judge the presence or absence of target substance, for positive rate up in 99.9%, false positive rate is less than 0.1%；(3), quickly, efficiently expand
Increase：Detection time 2 hours or so；(4), high sensitivity：Minimum detectability is 55CFU/mL；(5), identification is easy：Need to only it pass through
TMB developing solutions observation color change is added or multi-function microplate reader measures the judgement of OD values as a result, developing solution becomes after micropore is added
Blue and (bacterium solution OD values-blank control OD values)/(negative control OD value-blank control OD values) >=2.1 can be determined as the positive
(6), purposes is wide：It can be widely used for the safely and fast detection of SA.
Description of the drawings
Fig. 1：The sandwich DNA hybridization specific amplification test result of SA；
Fig. 2：The sandwich DNA hybridization of SA expands sensitivity test result；
Fig. 3：Yin and yang attribute compares sandwich DNA hybridization amplification；
In Fig. 1：1-17 is respectively：Pseudomonas aeruginosa, Enterobacter sakazakii, vibrio fluvialis, Shigella flexneri, enterocolitis
Ademilson bacterium, citrobacter freundii, Vibrio vulnificus, Klebsiella Pneumoniae, shigella dysenteriae, staphylococcus epidermis, saprophytic Portugal
Grape coccus, bacillus cereus, streptococcus fecalis, beta hemolytic streptococcus, Ying Nuoke Listerias, walsh staphylococcus, monokaryon
Monocytogenes；18：Staphylococcus aureus
In Fig. 2：1、5.5×107CFU/mL SA bacterium solutions, 2,5.5 × 106CFU/mL SA bacterium solutions, 3,5.5 × 105 CFU/
ML SA bacterium solutions, 4,5.5 × 104CFU/mL SA bacterium solutions, 5,5.5 × 103CFU/mL SA bacterium solutions, 6,5.5 × 102CFU/mL SA
Bacterium solution, 7,5.5 × 101CFU/mL SA bacterium solutions, 8,5.5CFU/mL SA bacterium solutions.
Specific implementation mode
Reaction solution without special instruction in the present invention, is commercial product.
Embodiment 1, the sandwich DNA hybridization technology quick detection kits of SA include one piece of 96 hole for being coated with poly dT
Plate, one bottle of pretreatment liquid, two bottles of dense bacterium solution lysates, one bottle of lysis buffer, one bottle of nucleic acid hybridization solution, one bottle of probe liquid,
One bottle of cleaning solution, one bottle of developing solution, one bottle of terminate liquid, one bottle of positive control solution and one bottle of negative controls, wherein：
Probe liquid consists of the following compositions：DNA sequence dna is the 16 μ L of SA capture probes of 6 μm of ol/L of SEQ ID NO.1；
DNA sequence dna is the 16 μ L of SA detection probes of 6 μm of ol/L of SEQ ID NO.2；Pretreatment liquid, volume are 30 μ L；Bacterium solution lysate,
Volume is 30 μ L；Nucleic acid hybridization solution, volume are 96 μ L；Cleaning solution uses after needing 10 times of dilutions, and volume is 750 μ L；Developing solution,
Volume is 150 μ L；Terminate liquid, volume are 100 μ L；It is the dosage of micropore reaction above.
The positive control solution, interior pipe is SA bacterium solutions, volume 5mL；
The negative controls, interior pipe is non-SA bacterium solutions, volume 5mL.
Embodiment 2, the design of probe
Staphylococcus aureus enterotoxin (SEs) is to cause the first cause of food poisoning, and heat stable nuclease can be with
Enterotoxin synthesizes under substantially the same conditions, exists in food that all can be after the heating, and in food storage and processing
Stability similar with enterotoxin can be kept under the conditions of various, the heat stable nuclease detected in food will be seen that golden yellow Portugal
The pollution condition of grape coccus and enterotoxin, to determine whether there is the possibility for causing food poisoning.Encode heat stable nuclease
Nuc genes are genes specific to staphylococcus aureus, and have higher conservative between different strains.Therefore root
According to the nuc gene reference sequences of the GenBank SA announced, multiple alignment, the conserved region of analytical sequence are carried out with Clustal W.
Using probe design software Primer 5, specific probe is designed, is respectively labeled as：SEQ ID NO.1, SEQ ID NO.2.Its
Capture probe in middle probe group:SEQ ID NO.1；Detection probe:SEQ ID NO.2；Volume ratio is 1:1；In capture probe 3 '
End label poly dA hold label horseradish peroxidase in detection probe 5 '；All probes are had by precious bioengineering (Dalian)
Limit company synthesizes.Probe sequence is as follows：
Embodiment 3, the preparation of positive reference substance
With pancreas peptone soybean broth (TSB) training status positive SA bacterium solutions 48h.Bacterium is measured using the method for plate culture count
The concentration of liquid makes the control of its concentration 1.0 × 107~1.0 × 108CFU/mL, one bottle of 5mL.
Embodiment 4, the preparation of negative controls
Escherichia coli 48h is cultivated with pancreas peptone soybean broth (TSB), takes out packing, one bottle of 5mL.
Embodiment 5, the bacterium solution template for preparing measuring samples
200 μ L or 200mg~300mg measuring samples of μ L~300 are taken, 48h is cultivated with pancreas peptone soybean broth (TSB),
As bacterium solution template.
The optimization of embodiment 6, hybridization conditions
With same concentrations staphylococcus aureus bacterium solution be detection object, respectively to probe liquid concentration (2 μM, 3 μM, 4 μM,
5 μM), probe hybridization solution proportioning (1:2、1:3、1:4、1:5), hybridization temperature (35 DEG C, 45 DEG C, 55 DEG C, 65 DEG C), hybridization time
(50min, 60min, 70min, 80min) designs the experiment of four factors, four horizontal quadrature, determines the optimal conditions of this detection method.Such as
Shown in embodiment 8.
Embodiment 7, specificity and sensitivity
Using the hybridization conditions of above-mentioned optimization, to standard positive staphylococcus aureus, Pseudomonas aeruginosa, Enterobacter sakazakii,
Vibrio fluvialis, Shigella flexneri, yersinia enterocolitica, citrobacter freundii, Vibrio vulnificus, Klebsiella Pneumoniae,
Shigella dysenteriae, staphylococcus epidermis, staphylococcus saprophyticus, bacillus cereus, streptococcus fecalis, beta hemolytic streptococcus,
The bacterium solutions such as Ying Nuoke Listerias, walsh staphylococcus, Listeria Monocytogenes are expanded, except golden yellow grape
Outside coccus, remaining is without non-specific hybridization, referring to Fig. 1.Absorbance such as following table in microplate reader at 450nm.
With sterile saline gradient dilution positive SA bacterium solutions, reacted using the hybridization conditions of above-mentioned optimization, this hair
Bright minimum detection limit respectively may be about 55CFU/mL.
The assembling of embodiment 8, kit
One piece to be coated with 96 orifice plates of poly dT, one bottle of pretreatment liquid, two bottles of dense bacterium solution lysates, one bottle of cracking slow
Fliud flushing, one bottle of nucleic acid hybridization solution, one bottle of probe liquid, one bottle of cleaning solution, one bottle of developing solution, one bottle of terminate liquid, one bottle of positive control
Liquid and one bottle of negative controls, stick date of manufacture and Product labelling.Cord blood and transport.
The sandwich DNA hybridization technology rapid detection method of embodiment 9, SA, includes the following steps：
(1) increase bacterium and cultivate SA bacterium solutions or measuring samples 48h with pancreas peptone soybean broth (TSB), obtain SA and detect bacterium solution
Or measuring samples liquid.
(2) SA is detected bacterium solution or measuring samples liquid and pretreatment liquid and lysate 4 ﹕, 1 ﹕ by volume by bacterium solution cracking reaction
1 ratio mixes in a glass tube, is put into 37 DEG C of water-baths and cultivates 5 minutes.
(3) bacterium solution that hybridization reaction is drawn after the cracking of 150 μ L steps (2) is added in the micropore for being coated with poly dT, then
The ratio mixed nucleus acid hybridization liquid of 3 ﹕ 1 and probe liquid (capture probe and detection probe are mixed with the ratio of 1 ﹕ 1) by volume, inhale
125 μ L are taken to be added in the micropore containing bacterium solution, at room temperature in being placed within 2 minutes with 150rpm speed rocker on rail mounted shaking table
70min is incubated in 45 DEG C of incubators.With 1 × PBST Buffer board-washings 5 times after incubation, add 150 μ L TMB developing solutions to each
It is incubated at room temperature in micropore 20 minutes, is eventually adding the sulfuric acid terminate liquid of 100 μ L 2M, is put into multi-function microplate reader and reads
Absorbance at 450nm.
(4) result judgement (bacterium solution OD values-blank control OD values)/(negative control OD value-blank control OD values) >=2.1
It can be determined as the positive, and become blue after developing solution addition micropore, color is deeper to illustrate that cause of disease bacteria concentration is higher.
Can be apparent from referring to Fig. 3+contain SA.Absorbance such as following table in microplate reader at 450nm.
SEQUENCE LISTING
<110>Chongqing Entry-Exit Inspection and Quarantine Bureau inspection and quarantine technique center
<120>The sandwich DNA hybridization of staphylococcus aureus, which quickly detects, uses probe, kit and detection method
<160> 2
<210> 1
<211> 22
<212> DNA
<213>Artificial sequence
<400> SEQ ID NO.1
gcgattgatg gtgatacggt ta 22
<210> 2
<211> 22
<212> DNA
<213>Artificial sequence
<400> SEQ ID NO.2
aatatggtcc tgaagcaagt gc 22
Claims (5)
1. the quick detection probe of the sandwich DNA hybridization of staphylococcus aureus, it is characterised in that：It is examined including SA capture probes and SA
Probing needle, the wherein DNA sequence dna of SA capture probes are that the DNA sequence dna of SEQ ID NO.1, SA detection probes is SEQ ID NO.2.
2. a kind of quick detection kit of sandwich DNA hybridization of staphylococcus aureus, it is characterised in that：Including being coated with poly
96 orifice plates of dT, pretreatment liquid, bacterium solution lysate, lysis buffer, nucleic acid hybridization solution, probe liquid, cleaning solution, developing solution, end
Only liquid, positive control solution and negative controls, the dosage that one of micropore reacts are：
The probe liquid consists of the following compositions：
DNA sequence dna is the 16 μ L of SA capture probes of 6 μm of ol/L of SEQ ID NO.1；
DNA sequence dna is the 16 μ L of SA detection probes of 6 μm of ol/L of SEQ ID NO.2；
Total 32 μ L；
The pretreatment liquid, volume are 30 μ L；
Lysate, volume are 30 μ L, are formed by bacterium solution lysate and lysis buffer mixed preparing；
The nucleic acid hybridization solution, volume are 96 μ L；
The cleaning solution uses after needing 10 times of dilutions, and volume is 750 μ L；
The developing solution, volume are 150 μ L；
The terminate liquid, volume are 100 μ L.
3. the quick detection kit of the sandwich DNA hybridization of staphylococcus aureus according to claim 2, it is characterised in that：
The positive control solution is SA bacterium solutions；The negative controls are non-SA bacterium solutions.
4. a kind of rapid detection method of the non-disease testing goal of the sandwich DNA hybridization of staphylococcus aureus, including walk as follows
Suddenly：
(1) increase bacterium and cultivate bacterium solution 48h to be measured with pancreas peptone soybean broth (TSB), obtain measuring samples liquid；
(2) bacterium solution cracking reaction is by measuring samples liquid, pretreatment liquid and lysate 4 ﹕ 1 by volume:1 ratio is in a glass tube
Mixing, is put into 37 DEG C of water-baths and cultivates 5 minutes；
(3) bacterium solution that hybridization reaction is drawn after the cracking of 150 μ L steps (2) is added in the micropore for being coated with poly dT, then presses volume
Than 3:1 ratio mixed nucleus acid hybridization liquid and probe liquid are drawn 125 μ L and are added in the micropore containing bacterium solution, at room temperature in track
It is placed in incubator within 2 minutes and is incubated with 150rpm speed rocker on formula shaking table；It is washed with 1 × PBST Buffer after incubation
Plate 5 times adds and is incubated at room temperature in 150 μ L developing solutions to each micropore 20 minutes, is eventually adding the terminate liquid of 100 μ L 2M, puts
Enter the absorbance read in multi-function microplate reader at 450nm；
(4) result judgement (bacterium solution OD values-blank control OD values)/(negative control OD value-blank control OD values) >=2.1 can be sentenced
It is set to the positive, and becomes blue after developing solution addition micropore, the more deep bacteria concentration to be measured of color is higher.
5. the sandwich DNA hybridization rapid detection method of staphylococcus aureus according to claim 4, it is characterised in that：It is described
Capture probe and detection probe are in the ratio mixing with 1 ﹕ 1 of volume ratio in probe liquid.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7662562B2 (en) * | 2004-08-10 | 2010-02-16 | Becton, Dickinson And Company | Method for rapid identification of microorganisms |
| CN102864226A (en) * | 2012-09-21 | 2013-01-09 | 武汉真福医药科技发展有限公司 | LAMP (loop-mediated isothermal amplification) kit for rapid detection of staphylococcus aureus |
| CN105063204A (en) * | 2015-08-07 | 2015-11-18 | 重庆出入境检验检疫局检验检疫技术中心 | Salmonella sandwich DNA hybridization rapid detection probe, kit, and detection method |
-
2018
- 2018-05-11 CN CN201810449424.4A patent/CN108546768A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7662562B2 (en) * | 2004-08-10 | 2010-02-16 | Becton, Dickinson And Company | Method for rapid identification of microorganisms |
| CN102864226A (en) * | 2012-09-21 | 2013-01-09 | 武汉真福医药科技发展有限公司 | LAMP (loop-mediated isothermal amplification) kit for rapid detection of staphylococcus aureus |
| CN105063204A (en) * | 2015-08-07 | 2015-11-18 | 重庆出入境检验检疫局检验检疫技术中心 | Salmonella sandwich DNA hybridization rapid detection probe, kit, and detection method |
Non-Patent Citations (1)
| Title |
|---|
| CARL A. BATT等: "《ENCYCLOPEDIA OF FOOD MICROBIOLOGY SECOND EDITION》", 31 December 2014, ACADEMIC PRESS ELSEVIER * |
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