CN106442987B - The fluorescence detection reagent kit and its application method of staphylococcus aureus - Google Patents
The fluorescence detection reagent kit and its application method of staphylococcus aureus Download PDFInfo
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56938—Staphylococcus
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Abstract
The invention discloses a kind of fluorescence detection reagent kit of staphylococcus aureus and its application method, kit includes following components：Staphylococcus aureus specific phage polypeptide coupled bead and fluoresceinisothiocyanate Magainin I, its application method is the addition staphylococcus aureus specific phage polypeptide coupled bead into sample to be tested, it is incubated for, Magnetic Isolation, fluoresceinisothiocyanate Magainin I is added, is incubated for, Magnetic Isolation, fluorescence signal is detected and recorded, the concentration of staphylococcus aureus in sample to be tested is acquired according to fluorescence intensity.The present invention is using the special phage polypeptide of staphylococcus aureus to be measured and broad spectrum antimicrobial peptide Magainin I as bacterium identification agent, in conjunction with magnetic bead fast enriching technology and highly sensitive fluorescence detection method, detected for the full cell of staphylococcus aureus, have the advantages that quickly, conveniently, specifically, sensitive, stable, low price.
Description
Technical field
The invention belongs to technical field of microbial detection, it is related to a kind of detection kit of staphylococcus aureus and its makes
Use method.
Background technique
Staphylococcus aureus is a kind of important pathogen of the mankind, is to cause the most common pathogen of pyogenic infection,
Including local pyogenic infection, systemic infection and systemic infection.Therefore, establish quickly, conveniently, sensitive golden yellow grape
Coccus detection method, for human health important in inhibiting.
In the existing detection method of staphylococcus aureus, the most mature is the tradition side based on Bacteria Culture mode
Method, most widely used method are the PCR methods based on bacterial nucleic acid.Although the conventional method reliability based on Bacteria Culture mode
It gets well, high sensitivity, but time and effort consuming, it usually needs the time of a couple of days can just obtain testing result, while need skilled profession
Personnel and a large amount of experimental apparatus.Although the PCR method specificity based on bacterial nucleic acid is high, multi-analyte immunoassay can be achieved, need
It will be by extracting the complicated process such as nucleic acid and synthesis specific primer.In addition, the method for detecting staphylococcus aureus
Southern blot hybridization technique and MTT method also based on DNA probe etc., although these method high specificities,
Sensitivity is very low, can not achieve the highly sensitive detection to staphylococcus aureus.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of fluorescence detection reagent kit of staphylococcus aureus and its making
With method, can quickly, conveniently, specifically, sensitive, stable, qurer detect staphylococcus aureus.
In order to achieve the above objectives, the present invention provides the following technical solutions：
1. the fluorescence detection reagent kit of staphylococcus aureus, including following components：Staphylococcus aureus specific is bitten
Thallus polypeptide coupled bead and fluoresceinisothiocyanate Magainin I.
Magainin I is the small peptide being made of in xenopous laevis skin particles gland 23 amino acid, is had to bacterium and fungi
Certain antibacterial activity, act on bacterium surface and have thermal stability, has a broad antifungal spectrum, be readily synthesized, that stability is good etc. is excellent
Point.
Preferably, the staphylococcus aureus specific phage polypeptide coupled bead is by avidin magnetic bead and biology
After the staphylococcus aureus specific phage polypeptide reaction of element modification, then is closed and be made with gelatin.
Preferably, the staphylococcus aureus specific phage polypeptide of the biotin modification is in Staphylococcus aureus
Modified biological is plain again after carbon teminal 3 glycine residues of connection of bacterium ATCC25923 specific bacteriophage polypeptide.
Preferably, the fluoresceinisothiocyanate Magainin I is the upper different sulphur of carbon teminal modification in Magainin I
Cyanic acid fluorescein.
In the above-mentioned technical solutions, according to the type for the staphylococcus aureus to be detected, the type golden yellow Portugal is selected
The phage polypeptide of grape coccus specificity prepares phage polypeptide coupled bead, can be realized to the type staphylococcus aureus
Detection.
As an optimal technical scheme, the staphylococcus aureus is staphylococcus aureus ATCC25923；It is described
Staphylococcus aureus specific phage polypeptide is staphylococcus aureus ATCC25923 specific bacteriophage polypeptide, ammonia
Base acid sequence is Val-Pro-His-Asn-Pro-Gly-Leu-Ile-Ser-Leu-Gln-Gly（SEQ ID No.1）.
It further, further include following components in the kit：Staphylococcus aureus ATCC25923 bacteria suspension, reaction
Buffer and detection buffer.
Preferably, the staphylococcus aureus ATCC25923 bacteria suspension is made by following methods：Take Staphylococcus aureus
Bacterium ATCC25923 single bacterium colony is inoculated in LB culture solution, 37 DEG C of shaken cultivations, and centrifugation removal supernatant, bacterium washes with water
Afterwards, it is resuspended with the phosphate buffer of 0.01 mol/L, pH 7.4 and adjusts bacterial concentration；The reaction buffer is 0.01
The phosphate buffer of mol/L, pH 7.4, the phosphate buffer that the detection buffer is 0.01 mol/L, pH8.0.
The application method of the fluorescence detection reagent kit of the staphylococcus aureus, be added into sample to be tested it is golden yellow
Color aureus specific phage polypeptide coupled bead, is incubated for, and it is more to obtain staphylococcus aureus-bacteriophage for Magnetic Isolation
Peptide coupled bead compound adds fluoresceinisothiocyanate Magainin I, is incubated for, and Magnetic Isolation obtains different sulphur cyanogen
Sour fluorescein is coupled Magainin I- staphylococcus aureus-phage polypeptide coupled bead compound, detects and records fluorescence
Signal acquires the concentration of staphylococcus aureus in sample to be tested according to fluorescence intensity.
As an optimal technical scheme, the user of the fluorescence detection reagent kit of staphylococcus aureus ATCC25923
Method includes the following steps：
(a) staphylococcus aureus is made in the dilution of staphylococcus aureus ATCC25923 bacteria suspension reaction buffer
ATCC25923 work bacterium solution；Staphylococcus aureus is added into staphylococcus aureus ATCC25923 work bacterium solution
ATCC25923 specific bacteriophage polypeptide coupled bead, is incubated for, and Magnetic Isolation obtains staphylococcus aureus ATCC25923-
Phage polypeptide coupled bead compound after being washed with reaction buffer, adds fluoresceinisothiocyanate Magainin
I is incubated for, Magnetic Isolation, obtains isothiocyanic acid coupling Magainin I- staphylococcus aureus ATCC25923- phage polypeptide
Coupled bead compound after being washed with reaction buffer, adds detection buffer, fluorescence signal is detected and record, with fluorescence
Intensity maps to staphylococcus aureus ATCC25923 concentration, draws working curve；
(b) staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled bead is added into sample to be tested,
It is incubated for, Magnetic Isolation obtains staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, is buffered with reaction
Liquid washing after, then plus fluoresceinisothiocyanate Magainin I, be incubated for, Magnetic Isolation, obtain fluorescein isothiocynate idol
Join Magainin I- staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, is washed with reaction buffer
After washing, detection buffer is added, detects and record fluorescence signal, the working curve drawn according to fluorescence intensity and step (a)
Acquire the concentration of staphylococcus aureus ATCC25923 in sample to be tested.
Preferably, the application method of the fluorescence detection reagent kit of staphylococcus aureus ATCC25923, including following step
Suddenly：
(a) by the dilution of staphylococcus aureus ATCC25923 bacteria suspension reaction buffer be made concentration 10 ~ 10 ×
105Staphylococcus aureus ATCC25923 work bacterium solution within the scope of CFU/mL；To 1.0 mL staphylococcus aureuses
10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled beads are added in ATCC25923 work bacterium solution, 37
DEG C be incubated for 45 minutes, Magnetic Isolation obtains staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, use
After reaction buffer washing, 5 μ g/mL fluoresceinisothiocyanate Magainin I 1.0mL are added, 37 DEG C are incubated for 45 points
It is more to obtain fluoresceinisothiocyanate Magainin I- staphylococcus aureus ATCC25923- bacteriophage for clock, Magnetic Isolation
Peptide coupled bead compound after being washed with reaction buffer, adds 200 μ L detection buffer, detects and record fluorescence letter
Number, it is mapped with fluorescence intensity to staphylococcus aureus ATCC25923 concentration, draws working curve；
(b) 10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptides are added into 1.0 mL samples to be tested
Coupled bead, 37 DEG C are incubated for 45 minutes, Magnetic Isolation, obtain staphylococcus aureus ATCC25923- phage polypeptide coupling magnetic
Pearl compound after being washed with reaction buffer, adds 5 μ g/mL fluoresceinisothiocyanate Magainin I 1.0mL, and 37
DEG C be incubated for 45 minutes, Magnetic Isolation obtains fluoresceinisothiocyanate Magainin I- staphylococcus aureus
ATCC25923- phage polypeptide coupled bead compound after being washed with reaction buffer, adds 200 μ L detection buffer,
Fluorescence signal is detected and recorded, golden yellow Portugal in sample to be tested is acquired according to the working curve that fluorescence intensity and step (a) are drawn
The concentration of grape coccus ATCC25923.
The beneficial effects of the present invention are：The present invention provides a kind of fluorescence detection reagent kit of staphylococcus aureus,
Using staphylococcus aureus specific phage polypeptide and broad spectrum antimicrobial peptide Magainin I as bacterium identification agent, in conjunction with magnetic
Pearl fast enriching technology and highly sensitive fluorescence detection method are detected for the full cell of staphylococcus aureus, are had fast
Speed, conveniently, specifically, sensitive, stables, low price the advantages of, be expected to the on-site test applied to staphylococcus aureus and quickly sieve
It looks into, staphylococcus aureus can be detected for fields such as clinical diagnosis, food safety and environment measurings and strong technology branch is provided
Maintain an equal level platform.
Detailed description of the invention
Fig. 1 is the use flow diagram of the fluorescence detection reagent kit of staphylococcus aureus ATCC25923.
Fig. 2 is to detect staphylococcus aureus using the fluorescence detection reagent kit of staphylococcus aureus ATCC25923
The working curve of ATCC25923.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, below in conjunction with attached drawing to of the invention excellent
Embodiment is selected to be described in detail.
The preparation of the fluorescence detection reagent kit of 1 staphylococcus aureus ATCC25923 of embodiment
The fluorescence detection reagent kit of the staphylococcus aureus ATCC25923 of the present embodiment, including following components：It is golden yellow
Staphylococcus A TCC25923 specific bacteriophage polypeptide coupled bead, fluoresceinisothiocyanate Magainin I, golden yellow
Staphylococcus A TCC25923 bacteria suspension, reaction buffer and detection buffer.
The preparation method of the staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled bead is：Take parent
It is mixed with biscuit porcelain pearl 2mg with the 30 μ g of staphylococcus aureus ATCC25923 specific bacteriophage polypeptide of biotin modification, instead
It answers, Magnetic Isolation, is washed with the phosphate buffer of 0.01 mol/L, pH 7.4, add 10 μ g/mL aqueous gelatin solutions
1.0mL closing, Magnetic Isolation, with the phosphate buffer of 0.01 mol/L, pH 7.4 wash to get.The golden yellow grape
The amino acid sequence of coccus ATCC25923 specific bacteriophage polypeptide is Val-Pro-His-Asn-Pro-Gly-Leu-Ile-
Ser-Leu- Gln-Gly（SEQ ID No.1）.The staphylococcus aureus ATCC25923 specificity of the biotin modification is bitten
Thallus polypeptide is after the carbon teminal of staphylococcus aureus ATCC25923 specific bacteriophage polypeptide connects 3 glycine residues
Modified biological element again.
The fluoresceinisothiocyanate Magainin I is that isothiocyanic acid is glimmering in the carbon teminal modification of Magainin I
Light element.
The preparation method of the staphylococcus aureus ATCC25923 bacteria suspension is：Take staphylococcus aureus
ATCC25923（From Guangdong Province, Culture Collection is obtained）Single bacterium colony is inoculated in LB culture solution, 37 DEG C of oscillation trainings
It supports 12 hours, centrifugation removal supernatant is slow with the phosphate of 0.01 mol/L, pH 7.4 after sterile water wash bacterium 2 times
Fliud flushing is resuspended bacterium and adjusts bacterial concentration to 1.0 × 105 CFU/mL。
The reaction buffer is the phosphate buffer of 0.01 mol/L, pH 7.4.
The phosphate buffer that the detection buffer is 0.01 mol/L, pH8.0.
The application method of the fluorescence detection reagent kit of 2 staphylococcus aureus ATCC25923 of embodiment
As shown in Figure 1, the use of the fluorescence detection reagent kit of staphylococcus aureus ATCC25923 prepared by embodiment 1
Method includes the following steps：
(a) by the dilution of staphylococcus aureus ATCC25923 bacteria suspension reaction buffer be made concentration 10 ~ 10 ×
105Staphylococcus aureus ATCC25923 work bacterium solution within the scope of CFU/mL；To 1.0 mL staphylococcus aureuses
10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled beads are added in ATCC25923 work bacterium solution, 37
DEG C be incubated for 45 minutes, Magnetic Isolation, obtain staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, use
Reaction buffer washing adds 5 μ g/mL fluoresceinisothiocyanate Magainin I 1.0mL, and 37 DEG C are incubated for 45 minutes,
Magnetic Isolation obtains fluoresceinisothiocyanate Magainin I- staphylococcus aureus ATCC25923- phage polypeptide
Coupled bead compound, is washed with reaction buffer, is added 200 μ L detection buffer, is detected and record fluorescence signal（Swash
Emission wavelength 488nm, wavelength of transmitted light 525nm）, mapped, drawn to staphylococcus aureus ATCC25923 concentration with fluorescence intensity
Working curve processed；
(b) 10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptides are added into 1.0 mL samples to be tested
Coupled bead, 37 DEG C are incubated for 45 minutes, Magnetic Isolation, obtain staphylococcus aureus ATCC25923- phage polypeptide coupling magnetic
Pearl compound, is washed with reaction buffer, adds 5 μ g/mL fluoresceinisothiocyanate Magainin I 1.0mL, and 37 DEG C
It is incubated for 45 minutes, Magnetic Isolation obtains fluoresceinisothiocyanate Magainin I- staphylococcus aureus ATCC25923-
Phage polypeptide coupled bead compound, is washed with reaction buffer, is added 200 μ L detection buffer, is detected and record glimmering
Optical signal acquires staphylococcus aureus in sample to be tested according to the working curve that fluorescence intensity and step (a) are drawn
The concentration of ATCC25923.
In above-mentioned steps (a), the concentration of staphylococcus aureus ATCC25923 work bacterium solution is respectively 10,1.0 × 102、
1.0×103、1.0×104With 1.0 × 105CFU/mL makees staphylococcus aureus ATCC25923 concentration with fluorescence intensity
Figure, gained working curve are as shown in Figure 2.With the increase of staphylococcus aureus ATCC25923 concentration, fluorescence intensity constantly increases
Greatly, the two is 10 ~ 1.0 × 105Good linear relationship, R are presented within the scope of CFU/mL2=0.9995, detection limit（S/N=3）For
12 CFU/mL illustrate delicately detect staphylococcus aureus ATCC25923 using detection kit of the invention.
In above-mentioned steps (a), 6 kinds of mode bacterium are separately added into staphylococcus aureus ATCC25923 work bacterium solution（Gold
Staphylococcus aureus CCTCC AB 91093, micrococcus luteus CCTCC AB 91100, staphylococcus epidermis GIM1.444, verdigris
Pseudomonas CCTCC AB 93078, Escherichia coli CCTCC AB 212355 and streptococcus ATCC35668）, then by above-mentioned
Same method is detected.The results show that other than staphylococcus epidermis GIM1.444 has weak interference, other 5 kinds of mode bacterium
Interference is not constituted to the detection of staphylococcus aureus ATCC25923, illustrating can be special using detection kit of the invention
Detect staphylococcus aureus ATCC25923 in strange land.
In above-mentioned steps (b), respectively using cider, lake water and Healthy People urine as sample to be tested, by its high-temperature sterilization
Afterwards, wherein cider and lake water do not dilute, and Healthy People urine spends ionized water and dilutes ten times, add the gold of known concentration thereto
Staphylococcus aureus ATCC25923 bacteria suspension, is then detected in same way as described above.The results are shown in Table 1, and 3 kinds to be measured
The recovery of standard addition of Gold Samples staphylococcus aureus ATCC25923 is that 81% ~ 105%, RSD value is not higher than 5.1%, illustrates use
Detection kit of the invention can accurately detect staphylococcus aureus ATCC25923.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although passing through ginseng
According to the preferred embodiment of the present invention, invention has been described, it should be appreciated by those of ordinary skill in the art that can
To make various changes to it in the form and details, without departing from the present invention defined by the appended claims
Spirit and scope.
<110>Southwest University
<120>The fluorescence detection reagent kit and its application method of staphylococcus aureus
<160> 1
<210> 1
<211> 12
<212> PRT
<213>Artificial sequence
<220>
<223>Staphylococcus aureus ATCC25923 specific bacteriophage polypeptide
<400> 1
Val Pro His Asn Pro Gly Leu Ile Ser Leu Gln Gly
1 5 10
Claims (6)
1. the fluorescence detection reagent kit of staphylococcus aureus, which is characterized in that including following components：Staphylococcus aureus is special
Anisotropic phage polypeptide coupled bead and fluoresceinisothiocyanate Magainin I；
The staphylococcus aureus specific phage polypeptide coupled bead is the gold by avidin magnetic bead and biotin modification
After staphylococcus aureus specific bacteriophage polypeptides reactive, then is closed and be made with gelatin；
The staphylococcus aureus specific phage polypeptide of the biotin modification is bitten in staphylococcus aureus specific
Modified biological is plain again after carbon teminal 3 glycine residues of connection of thallus polypeptide；
The fluoresceinisothiocyanate Magainin I is the upper fluorescein isothiocynate of carbon teminal modification in Magainin I；
The staphylococcus aureus is staphylococcus aureus ATCC25923；The staphylococcus aureus specific phagocytosis
Body polypeptide is staphylococcus aureus ATCC25923 specific bacteriophage polypeptide, amino acid sequence Val-Pro-His-
Asn-Pro- Gly-Leu-Ile-Ser-Leu-Gln-Gly。
2. the fluorescence detection reagent kit of staphylococcus aureus as described in claim 1, which is characterized in that in the kit
It further include following components：Staphylococcus aureus ATCC25923 bacteria suspension, reaction buffer and detection buffer.
3. the fluorescence detection reagent kit of staphylococcus aureus as claimed in claim 2, which is characterized in that the golden yellow Portugal
Grape coccus ATCC25923 bacteria suspension is made by following methods：Staphylococcus aureus ATCC25923 single bacterium colony is taken to be inoculated in LB
In culture solution, 37 DEG C of shaken cultivations, centrifugation removal supernatant, after bacterium washes with water, with the phosphorus of 0.01 mol/L, pH 7.4
Phthalate buffer is resuspended and adjusts bacterial concentration；The reaction buffer is the phosphate-buffered of 0.01 mol/L, pH 7.4
Liquid, the phosphate buffer that the detection buffer is 0.01 mol/L, pH8.0.
4. the application method of the fluorescence detection reagent kit of the described in any item staphylococcus aureuses of claims 1 to 3, feature
It is, staphylococcus aureus specific phage polypeptide coupled bead is added into sample to be tested, be incubated for, Magnetic Isolation obtains
To staphylococcus aureus-phage polypeptide coupled bead compound, fluoresceinisothiocyanate Magainin I is added,
It is incubated for, Magnetic Isolation, obtains fluoresceinisothiocyanate Magainin I- staphylococcus aureus-phage polypeptide coupling
Bead complexes detect and record fluorescence signal, and the concentration of staphylococcus aureus in sample to be tested is acquired according to fluorescence intensity.
5. the application method of the fluorescence detection reagent kit of staphylococcus aureus as claimed in claim 4, which is characterized in that packet
Include following steps：
(a) staphylococcus aureus is made in the dilution of staphylococcus aureus ATCC25923 bacteria suspension reaction buffer
ATCC25923 work bacterium solution；Staphylococcus aureus is added into staphylococcus aureus ATCC25923 work bacterium solution
ATCC25923 specific bacteriophage polypeptide coupled bead, is incubated for, and Magnetic Isolation obtains staphylococcus aureus ATCC25923-
Phage polypeptide coupled bead compound after being washed with reaction buffer, adds fluoresceinisothiocyanate Magainin
I is incubated for, Magnetic Isolation, obtains isothiocyanic acid coupling Magainin I- staphylococcus aureus ATCC25923- phage polypeptide
Coupled bead compound after being washed with reaction buffer, adds detection buffer, fluorescence signal is detected and record, with fluorescence
Intensity maps to staphylococcus aureus ATCC25923 concentration, draws working curve；
(b) staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled bead is added into sample to be tested, is incubated for,
Magnetic Isolation obtains staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, is washed with reaction buffer
After washing, then plus fluoresceinisothiocyanate Magainin I, be incubated for, Magnetic Isolation obtains fluoresceinisothiocyanate
Magainin I- staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, is washed with reaction buffer
Afterwards, detection buffer is added, fluorescence signal is detected and record, is asked according to the working curve that fluorescence intensity and step (a) are drawn
Obtain the concentration of staphylococcus aureus ATCC25923 in sample to be tested.
6. the application method of the fluorescence detection reagent kit of staphylococcus aureus as claimed in claim 5, which is characterized in that packet
Include following steps：
(a) concentration is made 10 ~ 10 × 10 in the dilution of staphylococcus aureus ATCC25923 bacteria suspension reaction buffer5
Staphylococcus aureus ATCC25923 work bacterium solution within the scope of CFU/mL；To 1.0 mL staphylococcus aureus ATCC25923
10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptide coupled beads are added in work bacterium solution, 37 DEG C are incubated for 45
Minute, Magnetic Isolation obtains staphylococcus aureus ATCC25923- phage polypeptide coupled bead compound, is buffered with reaction
After liquid washing, 5 μ g/mL fluoresceinisothiocyanate Magainin I, 1.0 mL is added, 37 DEG C are incubated for 45 minutes, magnetic
Separation obtains the coupling of fluoresceinisothiocyanate Magainin I- staphylococcus aureus ATCC25923- phage polypeptide
Bead complexes after being washed with reaction buffer, add 200 μ L detection buffer, fluorescence signal are detected and record, with glimmering
Luminous intensity maps to staphylococcus aureus ATCC25923 concentration, draws working curve；
(b) it is even that 10 μ g staphylococcus aureus ATCC25923 specific bacteriophage polypeptides are added into 1.0 mL samples to be tested
Join magnetic bead, 37 DEG C are incubated for 45 minutes, and Magnetic Isolation obtains staphylococcus aureus ATCC25923- phage polypeptide coupled bead
Compound after being washed with reaction buffer, adds 5 μ g/mL fluoresceinisothiocyanate Magainin I, 1.0 mL, and 37
DEG C be incubated for 45 minutes, Magnetic Isolation obtains fluoresceinisothiocyanate Magainin I- staphylococcus aureus
ATCC25923- phage polypeptide coupled bead compound after being washed with reaction buffer, adds 200 μ L detection buffering
Liquid detects and records fluorescence signal, is acquired according to the working curve that fluorescence intensity and step (a) are drawn golden yellow in sample to be tested
The concentration of staphylococcus A TCC25923.
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