CN109913527A - A method of utilizing shigella dysenteriae in ATP bioluminescence reaction detection food - Google Patents
A method of utilizing shigella dysenteriae in ATP bioluminescence reaction detection food Download PDFInfo
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Abstract
The invention discloses a kind of methods using shigella dysenteriae in ATP bioluminescence reaction detection food, centrifugation obtains initial supernatant liquid after this method first mixes sample to be detected with PBS buffer solution, digestive juice is then added and carries out heating in water bath for reaction, digestion supernatant is obtained after being centrifuged again and is filtered, filter membrane is immersed into mechanical shaking extraction in ATP release liquid, obtain ATP extracting solution, then the luminescence-producing reaction liquid being prepared is mixed in luminescence-producing reaction pipe with ATP extracting solution, it is placed in ATP fluorescence detector and detects luminous intensity, to realize the quick detection to shigella dysenteriae in food.Method easy quick, high sensitivity of the present invention using shigella dysenteriae in ATP bioluminescence reaction detection food can accurately detect to be greatly promoted the raising of food safety detection technology whether containing shigella dysenteriae and relative amount in food.
Description
Technical field
It is the present invention relates to biological detecting method technical field, in particular to a kind of to utilize ATP bioluminescence reaction detection food
The method of shigella dysenteriae in product.
Background technique
Shigella dysenteriae is also known as shigella dysenteriae, is a kind of Grain-negative bacillus pumilis, is that human bacterial's property dysentery is most commonly seen
Pathogen, Major Epidemic are generally called shigella dysenteriae in developing country, resist cold, and can pass through 24 hours on plain agar culture medium
Growth forms diameter up to 2 mm sizes, translucent smooth colony.Bacillary dysentery is the most common enteric infectious disease, summer
Two season of autumn, patient was most.The pathogenicity of shigella dysenteriae is stronger, and 10-100 bacterium of infection can fall ill.Shigella dysenteriae is in vegetables, melon
It can survive 1~2 week, and can breed in fruit, pickle, edible raw food food and unclean melon and fruit can cause bacillary dysentery.Therefore, quickly
Effectively the shigella dysenteriae in food is detected, so that the further diffusion of the infection sources is effectively controlled, for human health
It is of great significance with disease defence.
The conventional method of microorganism detection has medium therapy, PCR Molecular Detection method, enzyme linked immunosorbent assay and ATP biology
Luminescence method etc..Conventional medium method time-consuming is too long, and PCR molecular detecting method needs special instrument and equipment, need to specially extract
The DNA of sample, sample pre-treatments complex steps, operator need to have technical skill.Enzyme linked immunosorbent assay is made with antibody
To identify molecule, influence of the antibody vulnerable to external condition especially temperature requires harshness to storage conditions, largely
Limit the flexible Application of method.In addition, Antibody preparation is needed by zoopery or cell experiment, it is cumbersome bothersome, preparation
Antibody is at high cost, and the method testing cost of development is also corresponding higher.
ATP biloluminescence method is a kind of detection technique to grow up the 1980s, and this method is based on firefly
Principle of luminosity, under the catalytic action of luciferase, ATP and luciferin reaction discharge fluorescence, by test luminous intensity come real
The quick detection of existing microorganism.ATP is both the required substrate and the movable energy of all biological lifes of luciferase catalytic luminescence
Source is measured, the ATP content in every kind of microbial cell is constant.By the ATP concentration in measurement sample, work can be extrapolated
Bacterium number.It is to study a kind of more micro- life both at home and abroad at present because microculture, easy to operate, detection speed is fast without carrying out
Object detecting method.But for ATP biloluminescence method, the burst size of ATP will have a direct impact on test result in microorganism, if
ATP release not exclusively, then can make test result relatively low.And ATP is a kind of degradable energy matter, if release reagent can be broken
It is relatively low to also result in test result by bad ATP.Therefore, select suitable ATP release reagent most important for different test sources.
Summary of the invention
In view of the above-mentioned problems existing in the prior art and demand, it is sent out the object of the present invention is to provide a kind of using ATP biology
The method that light reaction detects shigella dysenteriae in food, this method are centrifuged after first mixing sample to be detected with PBS buffer solution
Initial supernatant liquid is obtained, digestive juice is then added and carries out heating in water bath for reaction, digestion supernatant is obtained after being centrifuged again and is filtered,
Filter membrane is immersed into mechanical shaking extraction in ATP release liquid, ATP extracting solution is obtained, then mentions the luminescence-producing reaction liquid being prepared with ATP
It takes liquid to mix in luminescence-producing reaction pipe, is placed in ATP fluorescence detector and detects luminous intensity, to realize to dysentery bar in food
The quick detection of bacterium.Method of the present invention using shigella dysenteriae in ATP bioluminescence reaction detection food is easy quickly,
High sensitivity can accurately detect to be greatly promoted food safety whether containing shigella dysenteriae and relative amount in food
The raising of detection technique.
Technical solution: to achieve the goals above, food is detected using ATP bioluminescence reaction the invention discloses a kind of
The method of middle shigella dysenteriae, comprising the following steps:
(1) the liquid/solid sample to be detected of 50 g is mixed with 1 × PBS buffer solution of 950 g, it is ground/to stir to get
Suspension/mixed liquor, is centrifuged off insoluble matter, and 200 μ L digestive juices are added into obtained initial supernatant liquid, are put into after mixing
15~20 min of digestion reaction is carried out in 40 degree of water-baths, is then centrifuged 1~2 min with 3000 g, is digested after abandoning precipitating
Supernatant；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and is then immersed in filter membrane and is contained with 50 mL
In the beaker of ATP release liquid, the ATP release liquid includes Triton X-100, trimethylamino methane, mercaptoethylmaine, 12
Sodium alkyl sulfate, saleratus, superoxide dismutase are then added 15 μ L apyrases, beaker are transferred to
40 DEG C of water-bath medium-rate mechanical shaking extractions, obtain ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, oscillation mix, and obtain luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 7.0~9.0 is added contains 0.13
The Tris-HCl buffering of~0.15 mol/L NaCl, 0.02~0.03 mol/L EDTA, 0.06~0.08 mol/L cyclodextrin
Total volume is complemented to 0.9 mL by liquid, adds the luminescence-producing reaction liquid of 0.1 mL step (3), is sufficiently placed in mixed liquor after oscillation
Luminous intensity is detected in ATP fluorescence detector, the detection response time is 20~30 s.
Preferably, the digestive juice composition in the step (1) are as follows: 0.1~0.3 mol/L Triton X-100,0.05~
0.07 mol/L disodium ethylene diamine tetraacetate, 0.02~0.04 mol/L Proteinase K, 0.5~0.9 mol/L dodecyl sulphur
Sour sodium.
Preferably, the pore size of filter membrane is 0.25~0.35 μm in the step (2).
Preferably, in the step (2) ATP release liquid composition are as follows: 0.04~0.06 mol/L Triton X-100,
0.05~0.07 mol/L trimethylamino methane, 0.03~0.05 mol/L mercaptoethylmaine, 2~4 mmol/L dodecanes
Base sodium sulphate, 13~15 mmol/L saleratus, 8~12 mmol/L superoxide dismutases.
Preferably, the rate of mechanical shaking extraction is 200 r/min in the step (2), and the time of mechanical shaking extraction is 8~10
min。
Preferably, soluble inorganic magnesium salts is any one in magnesium chloride, magnesium nitrate and magnesium sulfate in the step (3)
Kind is several.
Preferably, it is specifically with 10~20 min of rate oscillation of 150 r/min that the oscillation in the step (3), which mixes,.
Preferably, the pH of Tris-HCl buffer is 8.0 in the step (4).
Preferably, when mixed liquor is placed in ATP fluorescence detector the detection response for detecting luminous intensity in the step (4)
Between be 25 s.
Compared with prior art, the invention has the following beneficial effects:
(1) the method simplicity of the present invention using shigella dysenteriae in ATP bioluminescence reaction detection food is quick, sensitive
Degree is high, can accurately detect whether contain shigella dysenteriae and relative amount in food.
(2) method of the present invention using shigella dysenteriae in ATP bioluminescence reaction detection food, which is directed to, is detected
Special object shigella dysenteriae devise dedicated ATP release liquid, can for shigella dysenteriae carry out Specific lytic, sufficiently release
The ATP in shigella dysenteriae is put, the interference of ATP in other miscellaneous bacterias is avoided, substantially increases detection specificity.
Specific embodiment
It is of the invention below by way of combining following specific embodiments to further illustrate.It should be pointed out that real in detail below
It applies mode for explaining only the invention, is not used to be defined the contents of the present invention.
ATP release liquid provided by the invention, by by mercaptoethylmaine, lauryl sodium sulfate, saleratus, super oxygen discrimination
Change enzyme to be compounded with Triton X-100, the trimethylamino methane in tradition ATP release liquid, for shigella dysenteriae without bud
Born of the same parents have the characteristics of this Morphological Features of pili and its cell wall structure are Glanz negative bacterium without pod membrane, atrichia, majority,
It is anti-that chemistry is occurred with the thallus of shigella dysenteriae jointly by mercaptoethylmaine, lauryl sodium sulfate, saleratus, superoxide dismutase
It answers, the cell wall of shigella dysenteriae and cell membrane is made to decompose, to sufficiently discharge intracellular ATP, realize detection sensitivity
With the raising of accuracy.
Below by embodiment, the present invention is furture elucidated.It should be pointed out that the present invention is not intended to be limited to the reality
Apply example.
Embodiment 1
(1) it is mixed after being crushed the leaf of spinach to be detected of 50 g with 1 × PBS buffer solution of 950 g, it is agitated to be suspended
Liquid is centrifuged off insoluble matter, and 200 μ L digestive juices, digestive juice composition are as follows: 0.1 mol/L are added into obtained initial supernatant liquid
Triton X-100,0.05 mol/L disodium ethylene diamine tetraacetate, 0.02 mol/L Proteinase K, 0.5 mol/L dodecyl
Sodium sulphate is put into progress 15 min of digestion reaction in 40 degree of water-baths, is then centrifuged 1 min with 3000 g, abandons precipitating after mixing
After obtain digestion supernatant；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and the pore size of filter membrane is 0.25 μm, then will
Filter membrane is immersed in the beaker for being contained with 50 mL ATP release liquids, the composition of the ATP release liquid are as follows: 0.04 mol/L
Triton X-100,0.05 mol/L trimethylamino methane, 0.03 mol/L mercaptoethylmaine, 2 mmol/L dodecyls
15 μ L atriphos bis phosphoric acids are then added in sodium sulphate, 13 mmol/L saleratus, 8 mmol/L superoxide dismutases
Beaker is transferred to 40 DEG C of water-bath medium-rate mechanical shaking extractions by enzyme, and the rate of mechanical shaking extraction is 200 r/min, mechanical shaking extraction
Time is 8 min, obtains ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, is mixed with 10 min of rate oscillation of 150 r/min, obtains luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 8.0 is added contains 0.13 mol/
L NaCl, 0.02 mol/L EDTA, 0.06 mol/L cyclodextrin Tris-HCl buffer total volume is complemented into 0.9 mL,
The luminescence-producing reaction liquid of 0.1 mL step (3) is added, mixed liquor is placed in detection in ATP fluorescence detector sufficiently after oscillation and is shone
Intensity, detection response time are 25 s.
Embodiment 2
(1) the cold cuts sample to be detected of 50 g is mixed with 1 × PBS buffer solution of 950 g, it is ground to obtain suspension, from
The heart removes insoluble matter, and 200 μ L digestive juices, digestive juice composition are as follows: 0.2 mol/L are added into obtained initial supernatant liquid
Triton X-100,0.06 mol/L disodium ethylene diamine tetraacetate, 0.03 mol/L Proteinase K, 0.7 mol/L dodecyl
Sodium sulphate is put into progress 18 min of digestion reaction in 40 degree of water-baths, is then centrifuged 1.5 min with 3000 g after mixing, it is heavy to abandon
Digestion supernatant is obtained behind shallow lake；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and the pore size of filter membrane is 0.30 μm, then will
Filter membrane is immersed in the beaker for being contained with 50 mL ATP release liquids, the composition of the ATP release liquid are as follows: 0.05 mol/L
Triton X-100,0.06 mol/L trimethylamino methane, 0.04 mol/L mercaptoethylmaine, 3 mmol/L dodecyls
The double phosphorus of 15 μ L atriphos are then added in sodium sulphate, 14 mmol/L saleratus, 10 mmol/L superoxide dismutases
Beaker is transferred to 40 DEG C of water-bath medium-rate mechanical shaking extractions by sour enzyme, and the rate of mechanical shaking extraction is 200 r/min, mechanical shaking extraction
Time be 9 min, obtain ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, is mixed with 15 min of rate oscillation of 150 r/min, obtains luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 8.0 is added contains 0.14 mol/
L NaCl, 0.025 mol/L EDTA, 0.07 mol/L cyclodextrin Tris-HCl buffer total volume is complemented into 0.9 mL,
The luminescence-producing reaction liquid of 0.1 mL step (3) is added, mixed liquor is placed in detection in ATP fluorescence detector sufficiently after oscillation and is shone
Intensity, detection response time are 25 s.
Embodiment 3
(1) the salted & preserved vegetable pickling liquid to be detected of 50 g is mixed with 1 × PBS buffer solution of 950 g, it is agitated to be mixed
Liquid is centrifuged off insoluble matter, and 200 μ L digestive juices, digestive juice composition are as follows: 0.3 mol/ are added into obtained initial supernatant liquid
L Triton X-100,0.07 mol/L disodium ethylene diamine tetraacetate, 0.04 mol/L Proteinase K, 0.9 mol/L dodecane
Base sodium sulphate is put into progress 20 min of digestion reaction in 40 degree of water-baths, is then centrifuged 2 min with 3000 g after mixing, it is heavy to abandon
Digestion supernatant is obtained behind shallow lake；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and the pore size of filter membrane is 0.35 μm, then will
Filter membrane is immersed in the beaker for being contained with 50 mL ATP release liquids, the composition of the ATP release liquid are as follows: 0.06 mol/L
Triton X-100,0.07 mol/L trimethylamino methane, 0.05 mol/L mercaptoethylmaine, 4 mmol/L dodecyls
The double phosphorus of 15 μ L atriphos are then added in sodium sulphate, 15 mmol/L saleratus, 12 mmol/L superoxide dismutases
Beaker is transferred to 40 DEG C of water-bath medium-rate mechanical shaking extractions by sour enzyme, and the rate of mechanical shaking extraction is 200 r/min, mechanical shaking extraction
Time be 10 min, obtain ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, is mixed with 20 min of rate oscillation of 150 r/min, obtains luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 8.0 is added contains 0.15 mol/
L NaCl, 0.03 mol/L EDTA, 0.08 mol/L cyclodextrin Tris-HCl buffer total volume is complemented into 0.9 mL,
The luminescence-producing reaction liquid of 0.1 mL step (3) is added, mixed liquor is placed in detection in ATP fluorescence detector sufficiently after oscillation and is shone
Intensity, detection response time are 25 s.
Comparative example is (with bacteriolyze enzymes extraction mercaptoethylmaine, lauryl sodium sulfate, saleratus and super in ATP release liquid
Superoxide dismutase)
(1) the cold cuts sample to be detected of 50 g is mixed with 1 × PBS buffer solution of 950 g, it is agitated to obtain suspension, from
The heart removes insoluble matter, and 200 μ L digestive juices, digestive juice composition are as follows: 0.2 mol/L are added into obtained initial supernatant liquid
Triton X-100,0.06 mol/L disodium ethylene diamine tetraacetate, 0.03 mol/L Proteinase K, 0.7 mol/L dodecyl
Sodium sulphate is put into progress 18 min of digestion reaction in 40 degree of water-baths, is then centrifuged 1.5 min with 3000 g after mixing, it is heavy to abandon
Digestion supernatant is obtained behind shallow lake；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and the pore size of filter membrane is 0.30 μm, then will
Filter membrane is immersed in the beaker for being contained with 50 mL ATP release liquids, the composition of the ATP release liquid are as follows: 0.05 mol/L
15 μ L atriphos are then added in Triton X-100,0.06 mol/L trimethylamino methane, 0.5 g/L lysozyme
Beaker is transferred to 40 DEG C of water-bath medium-rate mechanical shaking extractions by bisphosphatase, and the rate of mechanical shaking extraction is 200 r/min, oscillation
The time of extraction is 9 min, obtains ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, is mixed with 15 min of rate oscillation of 150 r/min, obtains luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 8.0 is added contains 0.14 mol/
L NaCl, 0.025 mol/L EDTA, 0.07 mol/L cyclodextrin Tris-HCl buffer total volume is complemented into 0.9 mL,
The luminescence-producing reaction liquid of 0.1 mL step (3) is added, mixed liquor is placed in detection in ATP fluorescence detector sufficiently after oscillation and is shone
Intensity, detection response time are 25 s.
Reference examples
Embodiment 1-3 sample detected and comparative example sample detected are carried out according to conventional PCR Molecular Detection method flat
Row detection, to separately verify ATP bioluminescence reaction method used in ATP bioluminescence reaction method and comparative example used in the present invention
Detection accuracy.
Embodiment 1-3 is obtained into corresponding shigella dysenteriae sum through conversion with the fluorescence radiation intensity value of comparative example, specifically
Conversion method are as follows: in the luminous intensity of test sample, while shining for blank control is detected using sterile water as blank control
Intensity, sample luminous value take logarithm after subtracting blank luminous value, calculate the total number of bacteria in water sample to be measured by luminosity curve, carefully
Bacterium sum TBC=N × (505 × 10(A+BLogRLU))/3 × 10-16(a/liter), N=extension rate, RLU are relative light unit, and A, B are
After the standard ATP luminosity curve test of ATP luminescence-producing reaction liquid, ATP concentration and net relative light unit take the line obtained after double-log
Property return luminous equation coefficient.Obtained shigella dysenteriae sum is surveyed with the conventional PCR Molecular Detection method through reference examples
Fixed shigella dysenteriae sum is compared one by one, on the basis of the shigella dysenteriae sum that conventional PCR Molecular Detection method is measured,
The deviation percentage for the shigella dysenteriae sum that calculating embodiment 1-3 and comparative example are measured, the results are shown in Table 1.As it can be seen that of the invention
Method using shigella dysenteriae in ATP bioluminescence reaction detection food is only aided with the ATP release liquid of respective specific component
It is able to achieve high-precision detection effect, this is never to report in the prior art, to further increasing ATP bioluminescence reaction
The sensitivity of detection method plays a significant role.
Table 1
| Deviation percentage (%) | |
| Embodiment 1 | -1.95% |
| Embodiment 2 | -1.92% |
| Embodiment 3 | -1.93% |
| Comparative example | -7.47% |
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but can not
Therefore it is construed as limiting the scope of the patent.It should be pointed out that for those of ordinary skill in the art,
Under the premise of not departing from present inventive concept, various modifications and improvements can be made, and these are all within the scope of protection of the present invention.
Claims (9)
1. a kind of method using shigella dysenteriae in ATP bioluminescence reaction detection food, which is characterized in that including following step
It is rapid:
(1) the liquid/solid sample to be detected of 50 g is mixed with 1 × PBS buffer solution of 950 g, it is ground/to stir to get
Suspension/mixed liquor, is centrifuged off insoluble matter, and 200 μ L digestive juices are added into obtained initial supernatant liquid, are put into after mixing
15~20 min of digestion reaction is carried out in 40 degree of water-baths, is then centrifuged 1~2 min with 3000 g, is digested after abandoning precipitating
Supernatant；
(2) the digestion supernatant for taking 100 mL steps (1) to obtain is filtered, and is then immersed in filter membrane and is contained with 50 mL
In the beaker of ATP release liquid, the ATP release liquid includes Triton X-100, trimethylamino methane, mercaptoethylmaine, 12
Sodium alkyl sulfate, saleratus, superoxide dismutase are then added 15 μ L apyrases, beaker are transferred to
40 DEG C of water-bath medium-rate mechanical shaking extractions, obtain ATP extracting solution；
(3) by 10 μ L of luciferase, 30 mg of soluble inorganic magnesium salts, dithiothreitol (DTT) 15 mg and 50 mL in gnotobasis
Sterile water mixing, oscillation mix, and obtain luminescence-producing reaction liquid；
(4) for the ATP extracting solution that 0.2 mL step (2) of absorption obtains in luminescence-producing reaction pipe, pH 7.0~9.0 is added contains 0.13
The Tris-HCl buffering of~0.15 mol/L NaCl, 0.02~0.03 mol/L EDTA, 0.06~0.08 mol/L cyclodextrin
Total volume is complemented to 0.9 mL by liquid, adds the luminescence-producing reaction liquid of 0.1 mL step (3), is sufficiently placed in mixed liquor after oscillation
Luminous intensity is detected in ATP fluorescence detector, the detection response time is 20~30 s.
2. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, the digestive juice composition in the step (1) are as follows: 0.1~0.3 mol/L Triton X-100,0.05~0.07 mol/L
Disodium ethylene diamine tetraacetate, 0.02~0.04 mol/L Proteinase K, 0.5~0.9 mol/L lauryl sodium sulfate.
3. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, the pore size of filter membrane is 0.25~0.35 μm in the step (2).
4. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, the composition of ATP release liquid in the step (2) are as follows: 0.04~0.06 mol/L Triton X-100,0.05~0.07
Mol/L trimethylamino methane, 0.03~0.05 mol/L mercaptoethylmaine, 2~4 mmol/L lauryl sodium sulfate, 13
~15 mmol/L saleratus, 8~12 mmol/L superoxide dismutases.
5. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, the rate of mechanical shaking extraction is 200 r/min in the step (2), and the time of mechanical shaking extraction is 8~10 min.
6. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature exist
In soluble inorganic magnesium salts is any one or a few in magnesium chloride, magnesium nitrate and magnesium sulfate in the step (3).
7. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, it is specifically with 10~20 min of rate oscillation of 150 r/min that the oscillation in the step (3), which mixes,.
8. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature exist
In the pH of Tris-HCl buffer is 8.0 in the step (4).
9. the method according to claim 1 using shigella dysenteriae in ATP bioluminescence reaction detection food, feature
It is, mixed liquor, which is placed in ATP fluorescence detector, in the step (4) detects the detection response time of luminous intensity as 25 s.
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| CN113621140A (en) * | 2021-08-06 | 2021-11-09 | 江西师范大学 | Preparation method and application of enzyme-integrated rare earth coordination polymer |
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