CN104419644A - Luminous bacteria freeze-dried preparation and preparation method thereof - Google Patents
Luminous bacteria freeze-dried preparation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a method for preparing a luminous bacteria freeze-dried preparation. The method specifically comprises the following steps: a, providing a liquid mixture containing luminous bacteria and a protective agent; b, cooling the mixture to a first temperature interval T1, and maintaining the first temperature interval T1 for a period of time P1; c, heating the cooled mixture from the first temperature interval T1 to a second temperature interval T2, and maintaining the second temperature interval for a period of time P2; d, heating the heated mixture from the second temperature interval to a third temperature interval T3, and maintaining the temperature for a period of time P3; and e, drying the dormancy mixture, thereby forming the luminous bacteria freeze-dried preparation. The luminous bacteria freeze-dried preparation prepared by the method disclosed by the invention is high in stability, and the effective time of the activity after recovery is long.
Description
Technical field
The present invention relates to microbial preparation field, particularly, relate to the freeze-dried preparation field of photogenic bacterium especially Qinghai Vibrion (Vibrio qinghaiensis).
Background technology
Utilize photogenic bacterium for the susceptibility of poisonous substance, the composite acuity toxicity carrying out testing and evaluating environment or food has become more and more conventional monitoring method.
Utilizing photogenic bacterium test the technical testing of poisonous substance toxicity and evaluate in the composite acuity toxicity process of environment or food, usually use photogenic bacterium nutrient solution or photogenic bacterium freeze dried.
The nutrient solution of photogenic bacterium is compared with freeze dried, nutrient solution uses complicated, need first photogenic bacterium to be carried out the cultivation of solid slant culture base, culturing process often needs repeatable operation repeatedly, photogenic bacterium nutrient solution stability and consistence poor, require high to incubation time and environment and professional.
At present, photogenic bacterium freeze-dried preparation compares nutrient solution, although have advantage easy to carry, simple to operate, because the lyophilised bacteria validity hold-time after recovery is shorter, only 1 hours, can not meet long testing requirement.
Therefore, this area is high and can at recovery long-time effective freeze-dried preparation afterwards in the urgent need to developing a kind of stability.
Summary of the invention
The invention provides a kind of photogenic bacterium freeze-dried preparation and preparation method thereof, and a kind of test kit comprising photogenic bacterium freeze-dried preparation of the present invention.
A first aspect of the present invention, provides the preparation method of photogenic bacterium freeze-dried preparation, comprises step:
A. one is provided containing photogenic bacterium and protectant liquid form mixt;
B. described mixture be cooled to the first temperature range T1 and maintain for some time P1 in the first temperature range, thus forming the mixture after cooling, the first wherein said temperature range T1 is-100 ~-60 DEG C, is more preferably-90 ~-70 DEG C; P1 is 0.2 ~ 10 hour, preferably, is 0.5 ~ 8 hour, more preferably, is 1 ~ 4 hour;
C. the mixture after described cooling is warming up to the second temperature range T2 from the first temperature range T1, and maintain for some time P2 in the second temperature range, thus forming the rear mixture that heats up, the second described temperature range T2 is-55 ~-30 DEG C, is more preferably-40 ~-35 DEG C; P2 is 0.2 ~ 24 hour, more preferably, is 0.5 ~ 10 hour;
D. the mixture after described intensification is heated up and rise to the 3rd temperature range T3 from the second temperature range, and maintain for some time P3, thus form dormancy mixture, the 3rd described temperature range T3 is-5 ~ 0 DEG C, and P3 is 1 ~ 72 hour, more preferably, 5 ~ 24 hours are maintained; With
E. described dormancy mixture is carried out drying, thus form the freeze-dried preparation of photogenic bacterium.
In another preference, in stepb, described cooling is that 0 ~ 40 DEG C of liquid form mixt is down to the first temperature range T1.
In another preference, in stepb, described rate of temperature fall is 1-5 DEG C/min.
In another preference, in stepb, described temperature fall time is 0.25-2 hour.
In another preference, in steps d, also comprise step: the mixture after described intensification is warming up to stage by stage the 3rd described temperature range T3.
In another preference, described comprising stage by stage is divided into one-phase, two benches or three stages.
In another preference, comprise and described mixture is warming up to one or more sub-temperature range S, described sub-temperature range is selected from lower group :-25 ± 2 DEG C ,-20 ± 2 DEG C ,-15 ± 2 DEG C ,-10 ± 2 DEG C.
In another preference, being 5 ~ 60min at the Ps that holds time of each sub-temperature range, more preferably, is 10 ~ 30min.
In another preference, described is two benches stage by stage, and per stage temperature rise rate is 1 DEG C/min, and it is 15 minutes that per stage holds time.
In another preference, the speed of described intensification is 2-10 DEG C/min.
Preferably, the speed of described intensification is 3-8 DEG C/min.
In another preference, described method also has the one or more features being selected from lower group:
● the temperature rise rate being warming up to the second temperature range T2 from the first temperature range T1 is 1-6 DEG C/min;
● the heating-up time being warming up to the second temperature range T2 from the first temperature range T1 is 5-30 minute;
● the temperature rise rate being warming up to the 3rd temperature range T3 from the second temperature range T2 is 1-6 DEG C/min;
● the heating-up time being warming up to the 3rd temperature range T3 from the second temperature range T2 is 5-30 minute;
In another preference, described photogenic bacterium is selected from the photogenic bacterium with subordinate: Vibrio (Vibrio), Photobacterium (Photobacterium), Photorhabdus (Photorhabdus).
In another preference, described photogenic bacterium is Qinghai Vibrion or its derivative bacterium.
In another preference, described photogenic bacterium is the photobacterium phosphoreum from Photobacterium.
In another preference, described photogenic bacterium is the Fermi operator (Vibriofischeri) from Vibrio.
In another preference, the protective material described in step a comprises milk powder, trehalose
In step e, described drying is vacuum-drying.
A second aspect of the present invention, provides the freeze-dried preparation of a kind of photogenic bacterium, described freeze-dried preparation through recovery after, the survival transformation period T of photogenic bacterium
1/2>=4 hours.
In another preference, T
1/2for 5-10 hour.
In another preference, T
1/2>=5 hours, T
1/2>=6 hours, T
1/2>=7 hours, T
1/2>=8 hours, T
1/2>=10 hours,
A third aspect of the present invention, provides the freeze-dried preparation of a kind of photogenic bacterium, and in described freeze-dried preparation, the luminometer numerical value of photogenic bacterium is more than 400,000 number of photons.
In another preference, described luminometer numerical value for pressing method described in iso standard ISO11348-3-2007, measured value (I when latter 15 minutes of freeze-dried preparation recovery
k15or Ic
15).
In another preference, luminometer numerical value >=500,000 number of photons, >=600,000 number of photons or >=700,000 number of photons.
In another preference, described freeze-dried preparation is prepared by described method arbitrary in first aspect present invention.
A fourth aspect of the present invention, provides a kind of test kit, and described test kit comprises container and is arranged in the freeze-dried preparation described in the claim 7 or 8 of described container.
In another preference, described test kit also comprises the optional reagent that working instructions and optional one or more are selected from lower group:
(a) resuscitation fluid or recovery reagent;
(b) detection reagent;
In another preference, described resuscitation fluid comprises 0.85% sodium chloride solution
In another preference, described freeze-dried preparation and optional reagent are independent packagings.
A fifth aspect of the present invention, provides a kind of detection method, and described method comprises step:
I) freeze-dried preparation of the photogenic bacterium described in the present invention second or the third aspect is recovered, form the bacterium liquid of the photogenic bacterium after recovery;
Ii) the bacterium liquid of described photogenic bacterium is contacted with determinand, thus carry out the poisonous substance in qualitative or detection by quantitative determinand by the luminous intensity measuring photogenic bacterium.
In another preference, described determinand comprises water sample.
In another preference, described poisonous substance comprises heavy metal, agricultural chemicals, organic compound etc.
In another preference, described recovery step comprises and being mixed with resuscitation fluid by described freeze-dried preparation, thus also forms the bacterium liquid of the photogenic bacterium after recovery, and places for some time (as 0.5 ~ 2 hour) under room temperature (10 ~ 40 DEG C).
In another preference, described method also comprises step: the initial luminescence measuring the photogenic bacterium after recovery.
Should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and can combining mutually between specifically described each technical characteristic in below (eg embodiment), thus form new or preferred technical scheme.As space is limited, tiredly no longer one by one to state at this.
Accompanying drawing explanation
Fig. 1 shows in embodiment 1, prepares the temperature curve of Qinghai Vibrion freeze-dried preparation.
Fig. 2 is the result figure that reagent of the present invention does blank liquid test data.After can finding that the luminous inhibiting rate of reagent of the present invention steadily remains to 90 minutes always, there is no obvious downtrending.
Fig. 3 is the result figure using reagent of the present invention to do reference toxicant zinc chloride Example Test Data.After can finding that the luminous inhibiting rate of reagent of the present invention steadily remains to 22 hours always, there is no obvious downtrending.
The result figure of the blank liquid test data that Fig. 4 uses H reagent to do.Can find that the luminous inhibiting rate of H reagent has sharply downward trend after 60 minutes.
Embodiment
The present inventor is through extensive and deep research, be surprised to find that first, the photogenic bacterium bacterium liquid of cryopreservation is carried out hyperthermic treatment stage by stage, the quantity of photogenic bacterium in corresponding preparations can be improved extremely significantly, and the activity of photogenic bacterium and transformation period after recovery can be significantly improved (validity hold time length).On this basis, the present invention is completed.
Photogenic bacterium
Photogenic bacterium used in the present invention is not particularly limited, and can be any photogenic bacterium that can be applied to environment or the detection of sample poisonous substance.
" photogenic bacterium and environmental poisonous substance detect " ISBN978-7-5019-6625-7/X034 that the conventional visible red legend outstanding person of photogenic bacterium, Zheng Tianling, Li Weimin etc. write.
Typical photogenic bacterium comprises the photogenic bacterium be selected from subordinate: Vibrio (Vibrio), Photobacterium (Photobacterium), Photorhabdus (Photorhabdus) etc.Representational photogenic bacterium comprises (but being not limited to): Fermi operator (Vibrio fischeri), Qinghai Vibrion (Vibrio qinghaiensis), photobacterium phosphoreum (Photobacterium phosphoreum).
In another preference, described photogenic bacterium comprises Qinghai Vibrion, Fermi operator, photobacterium phosphoreum.
Photogenic bacterium freeze-dried preparation and preparation thereof
Photogenic bacterium freeze-dried preparation precision cv value provided by the invention is less than 10%, and the time of remaining valid after recovery is at least 8 hours, is more preferably 10 ~ 24 hours.
Photogenic bacterium freeze-dried preparation provided by the invention through recovery after, the medium effective concentration EC of photogenic bacterium
50t detection time
1/2>=4 hours.Preferably, T
1/2for 5-8 hour; More preferably T
1/2>=5 hours, T
1/2>=6 hours, T
1/2>=7 hours, T
1/2>=8 hours.
In order to prepare photogenic bacterium freeze-dried preparation of the present invention, must the strict temperature and time controlled in preparation process.Preparation method of the present invention can reduce for photogenic bacterium damage in freeze-drying process, and effectively improve the activity keeping photogenic bacterium and stability, concrete grammar is as follows:
A. one is provided containing photogenic bacterium and protectant liquid form mixt;
B. described mixture be cooled to the first temperature range T1 and maintain for some time P1 in the first temperature range, thus forming the mixture after cooling, the first wherein said temperature range is-100 ~-60 DEG C, is more preferably-90 ~-70 DEG C; P1 is 0.2 ~ 10 hour, preferably, is 0.5 ~ 8 hour, more preferably, is 1 ~ 4 hour;
C. the mixture after described cooling is warming up to the second temperature range T2 from the first temperature range T1, and maintain for some time P2 in the second temperature range, thus forming the rear mixture that heats up, the second described temperature range T2 is-55 ~-30 DEG C, is more preferably-40 ~-35 DEG C; P2 is 0.2 ~ 24 hour, more preferably, is 0.5 ~ 10 hour;
D. the mixture after described intensification is heated up and rise to the 3rd temperature range T3 from the second temperature range, and maintain for some time P3, thus form dormancy mixture, the 3rd described temperature range T3 is-5 ~ 0 DEG C, and P3 is 1 ~ 72 hour, more preferably, 5 ~ 24 hours are maintained; With
E. described dormancy mixture is carried out drying, thus form the freeze-dried preparation of photogenic bacterium.
Wherein, protective material comprises milk powder, trehalose etc.
Comprise and described mixture is warming up to one or more sub-temperature range S, described sub-temperature range is selected from lower group :-25 ± 2 DEG C ,-20 ± 2 DEG C ,-15 ± 2 DEG C ,-10 ± 2 DEG C.
In another preference, be 5 ~ 60min at the Ps that holds time of each sub-temperature range, more preferably, be that 10 ~ 30min photogenic bacterium of the present invention freeze-dried preparation preparation method can complete by artificial temperature control or by the freeze drying plant containing automatic temp sequence of control.
In another preference, the described freeze drying plant containing automatic temp sequence of control is program control cryogenic temperature freezing drying machine.
In another preference, described freeze drying plant is provided with pre-freezing temperature, is-100 ~-60 DEG C.
Test kit and using method
Test kit of the present invention comprises photogenic bacterium freeze-dried preparation of the present invention, solvent and sealed vessel, and wherein, described freeze-dried preparation and solvent are independently packaged in sealed vessel.
Solvent used in the present invention is not particularly limited, and photogenic bacterium freeze-dried preparation can be able to be made recover under room temperature (10-40 DEG C) and on photogenic bacterium and tested sample character without the medium affected for any.Typical solvent comprises 0.5% ~ 5% sodium chloride solution.
When using test kit of the present invention, sample and 17% sodium chloride solution can be mixed to form analyte mixed solution, and photogenic bacterium freeze-dried preparation provided by the invention and solvent are mixed to form luminescent bacteria mixed solution, after analyte mixed solution is become test fluid with luminescent bacteria mixed solution in the ratio remix of 19:1; Control group preparation process is the same, and difference is that analyte mixed solution is mixed by distilled water and 17% sodium chloride solution; Contrast liquid and test fluid are put into special photometer successively, obtain tested test fluid and contrast liquid reading, the velocity of variation that (method of calculation can reference standard: ISO11348-3-2007) obtain relative luminous intensity is calculated according to reading, the inhibiting rate of poisonous substance toxicity for photogenic bacterium luminescence is obtained according to the change of relative luminous intensity, thus successively according to the size (Luminous bacteria) judging poisonous substance toxicity.
Beneficial effect of the present invention
1. the photogenic bacterium freeze-dried preparation stability prepared of the inventive method is high: photogenic bacterium freeze-dried preparation precision cv value of the present invention is less than 10%, and the precision cv value of photogenic bacterium freeze-dried preparation prepared by traditional method is more than 20%, the two contrast has obvious difference.
2. the photogenic bacterium freeze-dried preparation prepared of the inventive method is long for the working lipe after recovery: photogenic bacterium freeze-dried preparation of the present invention is at least 8 hours working lipe, and the working lipe of photogenic bacterium freeze-dried preparation prepared by traditional method is 1 hours, therefore, freeze-dried preparation of the present invention can meet the test request of continuous a day substantially, and does not need repeatedly to prepare test formulation.
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.Unless otherwise indicated, otherwise per-cent and number are weight percent and parts by weight.
The preparation of embodiment 1 Qinghai Vibrion freeze-dried preparation:
Please sketch preparation method:
Separation Qinghai Vibrion bacterium liquid and protective material are mixed to form bacterium liquid mixture by 1.1;
1.2 freeze drying plants of mixture being inserted pre-freeze to-86 DEG C, and maintain 1 hour;
Mixture after cooling is warming up to-40 DEG C by 1.3, and maintains 2 hours, forms the mixture after once heating up;
Mixture after once heating up is warming up to-20 DEG C by 1.4 again, and maintains 15min, repeats this step, be warming up to-10 DEG C;
1.5 mixtures obtained 1.4 are warming up to 0 DEG C, and maintain 8 hours;
1.6 mixtures obtained 1.5, by vacuum-drying, obtain Qinghai Vibrion freeze-dried preparation.
The preparation of embodiment 2 Qinghai Vibrion freeze-dried preparation 2
Preparation method, with embodiment 1, refers to table 1:
The preparation of embodiment 3 Qinghai Vibrion freeze-dried preparation 3
Preparation method, with embodiment 1, refers to table 1:
Table 1
Embodiment 4 Qinghai Vibrion freeze-dried reagent stability test
Material: Qinghai Vibrion freeze dried (being called for short reagent of the present invention) prepared by the embodiment of the present invention 1.
Control group: pure water;
Experimental group: zinc chloride ZnCl
2, concentration is 0.6mg/L.
Testing method and step: adopt iso standard ISO11348-3-2007 " Water quality-Determination of the inhibitory effect of water samples on the lightemission of Vibrio fischeri (Luminescent bacteria test)-Part3Methodusing freeze-dried bacteria ", measure 15 minutes relative luminous intensity inhibiting rate H
t.
In table 2, I
0for luminous intensity at once prepared by reagent; I
k15for the luminous intensity after 15 minutes prepared by reagent; I
k15/ I
0represent the attenuation ratio of reagent luminous intensity, according to ISO11348-3-2007 regulation, this damping ratios should in the scope of 0.6 ~ 1.8; f
k15average is 0.8156.
In table 3, I
0for luminous intensity at once prepared by reagent; I
15for the luminous intensity of 15 minutes after reagent contact poison; I
c15for the luminous intensity after 15 minutes prepared by reagent; H
15be 15 minutes relative luminous intensity inhibiting rates;
Table 2
Table 3
The activity of embodiment 5 Qinghai Vibrion freeze-dried reagent is held time test
Material is with embodiment 4
Method selection: iso standard ISO11348-3-2007 " Water quality-Determination ofthe inhibitory effect of water samples on the light emission of Vibriofischeri (Luminescent bacteria test)-Part3Method using freeze-driedbacteria ".
Test duration selects: blank liquid contrast test is selected to be greater than 2 hours.Reference toxicant test selection is greater than 20 hours.
Test result is as Fig. 2 ~ Fig. 3:
Fig. 2 shows the result figure that reagent of the present invention does blank liquid test data.Can find, after the luminous inhibiting rate of reagent of the present invention steadily remains to 90 minutes always, there is no obvious downtrending.
Fig. 3 shows the result figure that reagent of the present invention does reference toxicant zinc chloride Example Test Data.Can find, after the luminous inhibiting rate of reagent of the present invention steadily remains to 22 hours always, there is no obvious downtrending.
The preparation of embodiment 6 photobacterium phosphoreum freeze-dried preparation
Preparation method is with embodiment 1.
Activity holds time experiment with embodiment 5, and comparative group luminous bacillus freeze-dried preparation can be obtained by conventional IQF method, refers to table 4:
The preparation of embodiment 7 Fermi operator freeze-dried preparation
Preparation method is with embodiment 1.
Activity holds time experiment with embodiment 5, and comparative group luminous bacillus freeze-dried preparation can be obtained by conventional IQF method, refers to table 4:
Table 4
Result shows, photobacterium phosphoreum prepared by the inventive method and the activity of Fermi operator freeze-dried preparation after recovery are held time and be all longer than freeze-dried preparation prepared by traditional method.
The Qinghai Vibrion freeze-dried reagent that comparative example 1 dry ice legal system is standby
Material and step, with reference to Chinese invention patent 97106203.X, are prepared into Qinghai Vibrion freeze-dried preparation (being called for short H reagent).
The Qinghai Vibrion freeze-dried preparation stability test of comparative example 2 comparative example 1 preparation
Material: Qinghai Vibrion freeze-dried preparation (H reagent) prepared by comparative example 1 of the present invention
Poisonous substance used and testing method are all with embodiment 4.
Test result is as shown in table 5 ~ 6:
Table 5
Table 6
From embodiment 4 and comparative example 2:
1. " the f in above-mentioned H reagent " blank group "
k15average " data of item are 0.4741; " f in reagent of the present invention " blank group "
k15average " data of item are 0.8156, according to 11th item item in iso standard ISO11348-3-2007 to the requirement of photogenic bacterium and evaluation, the stability of " reagent of the present invention " is better than " H reagent " greatly.
2. the data of " cv% " item in above-mentioned H reagent " blank group " are 20.42%; The data of " cv% " item in reagent of the present invention " blank group " are 3.42%, and the test precision that data show " reagent of the present invention " is compared " H reagent " and is greatly improved.
The activity of comparative example 3 Qinghai Vibrion freeze-dried reagent is held time test
Material: Qinghai Vibrion freeze-dried preparation (H reagent) prepared by comparative example 1 of the present invention
Method and test duration: with embodiment 5.
Test result is as shown in Figure 4:
Fig. 4 shows the result figure of H reagent set blank liquid test data.Can find, the luminous inhibiting rate of H reagent has sharply downward trend after 60 minutes.
Because H reagent is doing in the sample test of reference toxicant zinc chloride, completely cannot complete identical experiment, so there is no same embodiment 5 poisonous substance and testing identical data results.
Data according to embodiment 4,5 and comparative example 2,3 can be found out, maintain activity time (f after the level of activity after the recovery of freeze-dried preparation prepared by the present invention and recovery
k15average: 0.8156; Cv%:3.42%; Luminous inhibiting rate holds time>=and 22 hours) be all far superior to the standby H reagent (f of dry ice legal system
k15average: 0.4741; Cv%:20.42%; Luminous inhibiting rate is held time about 1 hour).
The all documents mentioned in the present invention are quoted as a reference all in this application, are just quoted separately as a reference as each section of document.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.
Claims (11)
1. a preparation method for photogenic bacterium freeze-dried preparation, is characterized in that, comprises step:
A. one is provided containing photogenic bacterium and protectant liquid form mixt;
B. described mixture be cooled to the first temperature range T1 and maintain for some time P1 in the first temperature range, thus forming the mixture after cooling, the first wherein said temperature range T1 is-100 ~-60 DEG C; P1 is 0.2 ~ 10 hour;
C. the mixture after described cooling is warming up to the second temperature range T2 from the first temperature range T1, and maintains for some time P2 in the second temperature range, thus form the rear mixture that heats up, the second described temperature range T2 is-55 ~-30 DEG C; P2 is 0.2 ~ 24 hour;
D. heated up by the mixture after described intensification and rise to the 3rd temperature range T3 from the second temperature range, and maintain for some time P3, thus form dormancy mixture, the 3rd described temperature range T3 is-5 ~ 0 DEG C, and P3 is 1 ~ 72 hour; With
E. described dormancy mixture is carried out drying, thus form the freeze-dried preparation of photogenic bacterium.
2. the method for claim 1, is characterized in that, also comprises step in steps d: the mixture after described intensification is warming up to stage by stage the 3rd described temperature range T3.
3. method as claimed in claim 2, it is characterized in that, described comprising stage by stage is divided into one-phase, two benches or three stages.
4. the method for claim 1, is characterized in that, the speed of described intensification is 2-10 DEG C/min, and more preferably, the speed of described intensification is 3-8 DEG C/min.
5. method as claimed in claim 2, it is characterized in that, described photogenic bacterium is selected from the photogenic bacterium with subordinate: Vibrio (Vibrio), Photobacterium (Photobacterium), Photorhabdus (Photorhabdus).
6. the method for claim 1, is characterized in that, the protective material described in step a comprises milk powder, trehalose; And/or
In step e, described drying is vacuum-drying.
7. a freeze-dried preparation for photogenic bacterium, is characterized in that, described freeze-dried preparation through recovery after, the survival transformation period T of photogenic bacterium
1/2>=4 hours.
8. a freeze-dried preparation for photogenic bacterium, is characterized in that, in described freeze-dried preparation, the luminometer numerical value of photogenic bacterium is more than 400,000 number of photons.
9. freeze-dried preparation as claimed in claim 7 or 8, is characterized in that, described freeze-dried preparation is prepared by described method arbitrary in claim 1 ~ 6.
10. a test kit, is characterized in that, described test kit comprises container and is arranged in the freeze-dried preparation described in the claim 7 or 8 of described container.
11. 1 kinds of detection methods, is characterized in that, described method comprises step:
I) freeze-dried preparation of the photogenic bacterium described in claim 7 or 8 is recovered, form the bacterium liquid of the photogenic bacterium after recovery;
Ii) the bacterium liquid of described photogenic bacterium is contacted with determinand, thus carry out the poisonous substance in qualitative or detection by quantitative determinand by the luminous intensity measuring photogenic bacterium.
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| CN107502567A (en) * | 2017-07-11 | 2017-12-22 | 深圳市朗石科学仪器有限公司 | Luminous bacillus freeze-dried powder and preparation method thereof |
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| CN107502567A (en) * | 2017-07-11 | 2017-12-22 | 深圳市朗石科学仪器有限公司 | Luminous bacillus freeze-dried powder and preparation method thereof |
| CN107502567B (en) * | 2017-07-11 | 2023-02-03 | 深圳市朗石科学仪器有限公司 | Photobacterium freeze-dried powder and preparation method thereof |
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