CN114540459A - Method for detecting heterogeneous drug resistance and stability of cyanobacteria antibiotics - Google Patents
Method for detecting heterogeneous drug resistance and stability of cyanobacteria antibiotics Download PDFInfo
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- 206010059866 Drug resistance Diseases 0.000 title claims abstract description 61
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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
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a method for detecting heterogeneous drug resistance and stability of cyanobacteria antibiotics, wherein the heterogeneous drug resistance of the cyanobacteria antibiotics comprises the steps of respectively culturing target species samples by adopting antibiotics with gradient concentration, respectively measuring the minimum antibiotic inhibition concentration of each target species sample by a microdilution method, and taking the lowest value as the community MIC of the target species; respectively culturing the blue algae to be tested by taking antibiotics with the concentrations of 0, 1 and n times of the colony MIC, wherein n is more than or equal to 2 and less than or equal to 8, counting the number of cells of each group of blue algae, and respectively recording the number as P0、P1、Pn(ii) a If Pn＜(10％～50％)P0Calculating the antibiotic heterogeneous drug resistance level f of the blue algae to be detected by the following formula:
the invention effectively reduces the blue algae group containing a small amount of drug-resistant sub-floraThe false negative false judgment probability avoids the underestimation of the traditional Minimum Inhibitory Concentration (MIC) test method on the antibiotic resistance level.
Description
Technical Field
The invention relates to a method for detecting drug resistance and stability thereof, in particular to a method for detecting heterogeneous drug resistance and stability of a blue algae antibiotic.
Background
In recent years, the detection rate of antibiotics in water bodies of rivers and lakes is continuously increased due to the lack of effective supervision on mass production, excessive use and disposal and discharge of the antibiotics, and the water bodies of the rivers and lakes become huge antibiotic and drug-resistant gene banks. The long-term exposure of the low-concentration antibiotics inevitably promotes the diffusion and the spread of the drug resistance of bacteria in modes of gene mutation induction, horizontal transmission and intergenerational inheritance, and the antibiotic resistance genes are genetically amplified through an aquatic food chain and are transmitted to edible aquatic products such as crustaceans and fishes step by step, so that the disease control effect of aquaculture is reduced, and the drug resistance of bacteria, particularly pathogenic bacteria, is promoted to be further increased through drinking water, the edible aquatic products and agricultural products to return to the human body, so that the treatment efficacy of the antibiotics is gradually reduced or even disappeared, the medical treatment time is prolonged, the cost is increased, and the drug resistance can be avoided or even the life risk can occur, thereby seriously threatening the health and social stability of human beings.
The blue-green algae is also called cyanobacteria, has similar cell structure characteristics and genetic characteristics with gram-negative bacteria, is an important indicator species for water ecosystem balance and water environment evolution, is also an important carrier and a transmission transfer station for antibiotic drug resistance genes in a water ecosystem, and can be used as an important basis for evaluating the pollution levels of the antibiotic and the drug resistance genes in the water body of rivers and lakes. The phenotype and genotype drug resistance of the blue-green algae is widely detected, the blue-green algae shows low sensitivity to some antibiotics through self-resistance, gene mutation under antibiotic pressure, exogenous drug resistance gene transduction and other modes, and the phenotype and genotype drug resistance of the blue-green algae usually shows obvious positive correlation. However, some cyanobacteria samples exhibit a high level of genotype resistance and at the same time often exhibit phenotypic susceptibility to antibiotics in the Minimum Inhibitory Concentration (MIC) test. Currently, the Minimum Inhibitory Concentration (MIC) is widely recognized by the academia as a criterion for bacterial sensitivity or drug resistance. MIC data are typically obtained under conditions conducive to antibiotic sterilization or bacteriostatic action, and are tested for low bacterial growth density, exponential growth phase on the medium, and short antibiotic addition time. However, pathogenic bacteria classified as susceptible based on MIC criteria may exhibit resistance after prolonged exposure to antibiotics, one of the important reasons being bacterial antibiotic heteroresistance.
Antibiotic resistance (antibiotic heteroresistance) generally refers to a single isolated strain in which a subpopulation of cells having different sensitivities to antibiotics is present in the cultured population, i.e., some cells are sensitive to antibiotics while other cells are resistant, and the bacterium is referred to as an antibiotic heteroresistant strain. The number of cyanobacterial cells of different subgroups sensitive to antibiotics is small and is difficult to reflect by MIC data, however, under the long-term antibiotic selection, the subgroups which are small are likely to become main groups, and unstable heterogeneous drug resistance can further aggravate the hiding degree of cyanobacterial antibiotic drug resistance false negative. The reliability of the MIC data of the blue-green algae strains in a laboratory is reduced by the heterogeneous blue-green algae resistance, the antibiotic resistance level of the river and lake water environment can be underestimated, and a severe challenge is brought to the antibiotic resistance evaluation of the blue-green algae environment samples.
Although the phenomenon of antibiotic-resistant heterosis has been discovered by relevant studies at home and abroad, the frequency of occurrence of bacterial heteroresistance to clinically used antibiotics and their relative contribution to the failure of antibiotic therapy are still poorly understood. The current research on the heterogeneous drug resistance of the cyanobacteria antibiotics is relatively lack, mainly because of two reasons, one is fuzzy of the heterogeneous drug resistance of the cyanobacteria antibiotics and the stability definition and identification methodology thereof, the other is that the MIC breakpoint of the cyanobacteria strains does not have an accurate database at present, and the method for clinically testing the antibiotic sensitivity is still very difficult to be used for detecting the heterogeneous drug resistance of the cyanobacteria.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics, which is efficient and economical and can reduce the false negative misjudgment probability; the invention also aims to provide a method for detecting the stability of the heterogeneous resistance of the cyanobacteria antibiotics.
The technical scheme is as follows: the invention discloses a method for detecting heterogeneous drug resistance of cyanobacteria antibiotics, which comprises the following steps:
(1) collecting N parts of blue-green algae community samples at different sampling points, separating blue-green algae target species from the blue-green algae community samples, and obtaining M parts of target species samples; wherein M is less than or equal to N, and N parts of blue algae communities comprise a plurality of blue algae species, but each part of blue algae community does not comprise a target species, so that only M parts of samples can be obtained by separating N parts of samples.
(2) And respectively culturing M target species samples by adopting antibiotics with gradient concentrations, respectively measuring the minimum inhibitory concentration of the antibiotics of each target species sample by a microdilution method, and taking the lowest value as the community MIC of the target species.
(3) Respectively culturing the blue algae to be tested by taking antibiotics with the concentrations of 0, 1 and n times of the MIC of the community, wherein the blue algae to be tested is the blue algae with a single species, the species is a target species, n is more than or equal to 2 and less than or equal to 8, counting the number of cells of each group of blue algae after culturing for the same days under the same culture conditions as the culture conditions in the step (2), and respectively recording the number as P0、P1、Pn(ii) a If Pn＜(10％～50％)P0Then, the resistance of the blue algae to be detected is calculated by the following formulaLevel of antibiotic heterogeneous resistance f:
further, in the step (3), if f is more than or equal to 0.0001-0.00001%, the blue algae to be detected has heterogeneous drug resistance; f is less than or equal to 0.0001-0.00001%, and the blue algae to be detected has sensitivity.
Further, in step (3), if P is present1≥P0And P isn≥P0The blue algae to be detected has drug resistance.
Further, in the step (1), separating and purifying the blue algae target species from the blue algae colony sample under the aseptic condition, culturing the blue algae target species in a BG11 liquid culture medium for several days, and concentrating until the density of blue algae cells is about 108-109The samples were expressed in terms of counts/L.
Further, in the step (2), Escherichia coli K12 for testing the activity of the antibiotic standard solution was added to the positive control group in the microdilution method.
Further, in the step (2), the method for judging the minimum inhibitory concentration of the antibiotic is as follows: and (4) compared with a negative control group, the antibiotic concentration completely inhibits the growth of the blue algae cells and destroys the blue algae cells.
Further, the number of cyanobacterial cells and the integrity of the cells were analyzed by densitometric analysis and fluorescence microscopy.
Further, in the step (2) and the step (3), the culture conditions were 25.0. + -. 0.5 ℃ and 50. + -. 5. mu.E/m/s, and the light-dark ratio was 14/10 hours.
On the other hand, the method for detecting the stability of the antibiotic heterogeneous drug resistance of the blue algae comprises the detection method according to any one of claims 1 to 8, and further comprises the following steps:
(1) separating the blue algae with antibiotic heterogeneous drug resistance obtained by screening to obtain a plurality of blue algae colonies, and culturing the blue algae colonies for a plurality of hours by taking the antibiotic with the concentration being n times of the MIC value of the colonies;
(2) measuring the MIC value of the original cyanobacteria colony of one part of the cultured cyanobacteria colony; transferring another part of cyanobacteria colony for more than or equal to 50 times in an environment without antibiotic pressure, and then measuring the MIC value of the transferred cyanobacteria colony; and if the MIC value of the transferred cyanobacteria colony is not obviously reduced compared with the MIC value of the original cyanobacteria colony, the stability of antibiotic heterogeneous drug resistance is realized.
Further, the method for transferring in the step (2) is to add the mother liquor of the blue algae colony to 50 times of the liquid culture medium of the sterile BG11 without adding antibiotics.
The invention establishes an efficient and economic method for detecting and judging the heterogeneous drug resistance and the stability of the cyanobacteria antibiotics, is used for replacing the traditional MIC test method, can avoid the underestimation of the MIC test method on the drug resistance level of the antibiotics, effectively reduces the false negative misjudgment probability of the cyanobacterial community containing a small amount of drug-resistant sub-flora, is beneficial to comprehensively and accurately evaluating the antibiotic drug resistance of the river and lake water ecological system, and has important scientific value for deepening the research on the water environment behavior mechanism of the antibiotic drug-resistant gene and the ecological risk control strategy.
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages: (1) the false negative misjudgment probability of the cyanobacterial community containing a small amount of drug-resistant sub-flora is effectively reduced, the underestimation of the traditional Minimum Inhibitory Concentration (MIC) test method on the antibiotic resistance level is avoided, the antibiotic MIC break point is replaced by the minimum inhibitory concentration of the cyanobacterial overall community, the technical bottleneck that the cyanobacterial antibiotic MIC break point lacks data is solved, the definition and identification method of the heterogeneous drug resistance of the cyanobacterial antibiotic is provided for the first time, and the analysis method and the judgment standard of the heterogeneous drug resistance level and the stability are defined; (2) the method is simple to operate, saves time, effectively improves the accuracy of the detection of the phenotypic resistance of the antibiotics of the blue algae samples, is beneficial to comprehensively and accurately evaluating the antibiotic resistance of the river and lake water ecological system, has important scientific value for deepening the research of the water environment behavior mechanism of antibiotic resistance genes and ecological risk control strategies, and has wide application prospect.
Drawings
FIG. 1 is a flowchart of determination of the level of heterogeneous resistance of cyanobacteria antibiotics;
FIG. 2 shows the result of determining the level of heterogeneous resistance of cyanobacteria antibiotics;
FIG. 3 is a flowchart of determination of the stability of heterogeneous resistance of cyanobacterial antibiotics;
FIG. 4 shows the result of determining the stability of the heterogeneous drug resistance of the cyanobacteria antibiotics.
Detailed Description
The technical scheme of the invention is further explained by combining the attached drawings.
Example 1
A method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics comprises the following steps:
(1) preparation of to-be-detected algal strain and antibiotic standard solution
Collecting blue algae samples: in Taihu lake, Hongze lake, nido lake and Dian lake water areas, 50 sampling points are arranged in total according to the comprehensive, representative, objective, feasible and continuous principles, and 50 blue algae community samples are collected.
Separating and culturing target species of the blue algae: separating and purifying 50 collected blue algae community samples, and totally separating 300 blue algae single communities, wherein the method comprises the following steps: anabaena, synechocystis, cyanobacteria, microcystis, oscillatoria, flounder, cisternaria, synechocystis, and synechocystis, wherein there are 14 anabaena, 25 synechocystis, 9 cyanobacteria, 182 microcystis, 16 oscillatoria, 7 flounder, 13 cisternaria, 19 synechocystis, and 15 synechocystis, and the culture is performed according to the species;
the culture method comprises the following steps: culturing single blue algae species in BG11 liquid culture medium for 5-7 days (25.0 + -0.5 deg.C, 50 + -5 μ E/m/s, 14/10 hr light-dark ratio), and concentrating until the density of blue algae cells is about 108-109one/L, 100. mu.L per well, was batched into 96-well plates.
Antibiotics: seven 19 common antibiotics are selected, including aminoglycosides, chloramphenicol, beta-lactams, macrolides, quinolones, sulfonamides and tetracyclines, and the specific antibiotic types are shown in table 1;
TABLE 1
Preparing standard solution: BG11 liquid culture medium was used to prepare antibiotic standard solutions of different concentrations, the gradient interval of the antibiotic standard solution concentration was 2 times.
(2) Determination of minimum inhibitory concentration of integral colony of blue-green algae
The target species of the blue algae is anabaena, and the antibiotic is gentamicin of aminoglycoside.
Respectively measuring the MICs of 14 anabaena on gentamicin by adopting an improved microdilution method; and (2) filling 100 mu L of single anabaena algae liquid into each hole in a 96-hole plate, then respectively filling 100 mu L of the gentamicin standard liquid prepared in the step (1) into the algae liquid to be tested of the 96-hole plate, wherein the algae liquid is not added in a blank control group, and only the antibiotic standard liquid is added in a negative control group, and the escherichia coli K12(ATCC 47076) is added in a positive control group for testing the activity of the antibiotic standard liquid. The 96-well plate is cultured for 14 days under the same conditions (25.0 +/-0.5 ℃, 50 +/-5 mu E/m/s, light-dark ratio of 14/10 hours), the number of blue algae and the integrity of cells are analyzed through optical density analysis (450nm) and fluorescence microscopy (400-1000 times), and compared with a negative control group, the minimum antibiotic concentration which completely inhibits the growth of blue algae cells and destroys the blue algae cells is calculated as MIC.
And repeating the steps until 14 Anabaena MICs for gentamicin are obtained, and taking the lowest value as the community MIC.
(3) Blue algae antibiotic bacteria spectrum analysis
Selecting any sampling point in the water areas of the Taihu lake, the Hongze lake, the nido lake and the Dian lake in the step (1) for blue algae sampling, separating a single anabaena from the sampling point as an algae solution to be detected, culturing for 5-7 days (25.0 +/-0.5 ℃, 50 +/-5 mu E/m/s and 14/10 hours light-dark ratio) in a BG11 liquid culture medium, and concentrating until the cell density of the blue algae is about 108-109And (2) per liter.
Preparing standard solutions with different antibiotic concentrations by using a BG11 liquid culture medium, wherein the antibiotic standard solutions are respectively 0 (negative control), 0.125, 0.25, 0.5, 1.0, 2.0, 4.0 and 8.0 times of the colony MIC, each standard solution is respectively added into 100 mu L of to-be-detected algae solution of a 96-well plate, and the setting of a control group, the culture conditions and the index detection method are the same as those in step 2); wherein the antibiotic standard solution concentration is 1.0, 2.0, 4.0 and 8.0 times of the colony MIC, and the antibiotic standard solution is called as antibiotic concentration group.
(4) Determination of heterogeneous drug resistance level of blue algae antibiotics
As shown in figure 1, the number of the blue algae population cells in different antibiotic concentration groups is analyzed to judge the heterogeneous drug resistance level of the blue algae antibiotics. In the antibiotic concentration group, if the number of the blue algae population cells is equal to or more than 10-50% of that of the negative control group, the drug resistance can be judged; if the blue algae population does not meet the drug resistance condition and the number of the subset cells in the blue algae population is equal to or higher than 0.0001-0.00001 percent of the negative control group in an antibiotic concentration group 8 times the MIC of the community, the blue algae population can be judged to have heterogeneous drug resistance; if the blue algae population does not meet the conditions of drug resistance and heterogeneous drug resistance, the blue algae population can be judged to have sensitivity.
The method for calculating the heterogeneous drug resistance level (f) of the blue algae antibiotics comprises the following steps:
wherein, PnThe number of blue algae population cells in an antibiotic concentration group 8 times the MIC of the population, i.e., the number of subset cells, P, in a heterogeneous drug-resistant blue algae population0The number of the blue algae population cells is a negative control group.
And (3) determining 14 blue algae target species and 19 antibiotics in a free combination manner, and repeating the steps (2) to (4) to obtain the heterogeneous drug resistance level of the blue algae antibiotics of 5700 different antibiotics and the blue algae strains to be determined in a combined manner.
As shown in FIG. 2, in 5700 different antibiotics and cyanobacteria strain combinations to be tested, 3568 combinations have drug resistance, 1137 combinations have heterogeneous drug resistance, and 995 combinations have sensitivity. Of the 1137 combinations with heterogeneous resistance, 896 showed resistance in the traditional MIC test and 241 showed sensitivity in the traditional MIC test.
Example 2
The method for judging the heterogeneous drug resistance stability of the cyanobacteria antibiotics comprises the following steps:
(1) and (3) re-separating the resistance subset cells obtained by screening the heterogeneous drug-resistant group, selecting 3-5 cyanobacteria colonies, re-inoculating the cyanobacteria colonies into a sterile BG11 liquid culture medium with the same antibiotic concentration, and continuously culturing for 72 hours. The culture conditions are 25.0 + -0.5 deg.C, 50 + -5 μ E/m/s, and 14/10 hr light-dark ratio.
(2) Performing MIC test on the cyanobacteria colonies, wherein the setting of a control group, the culture conditions and the index detection method are the same as the step (2) in the example 1; the subculture was performed 50 times in succession, 1mL of each stock solution was added to 50mL of sterile BG11 liquid medium without antibiotic addition, and the MIC value of the whole population of cyanobacteria was determined (as shown in fig. 3). If the cyanobacteria is transformed for 50 times in the environment without antibiotic pressure, the MIC value of the whole population of the cyanobacteria is obviously reduced, and the stability of the antibiotic heterogeneous drug resistance of the cyanobacteria can be judged.
As shown in fig. 4, of the 1137 antibiotics with heterogeneous resistance and cyanobacterial strain combinations, 97.7% of the combinations had no stability against heterogeneous resistance, and 2.3% of the combinations had stability against heterogeneous resistance.
Claims (10)
1. A method for detecting heterogeneous drug resistance of a cyanobacteria antibiotic is characterized by comprising the following steps:
(1) collecting a plurality of blue-green algae community samples at different sampling points, and respectively separating and purifying blue-green algae target species in the blue-green algae community samples to obtain target species samples;
(2) respectively culturing target species samples by adopting antibiotics with gradient concentrations, respectively measuring the minimum inhibitory concentration of the antibiotics of each target species sample by a microdilution method, and taking the lowest value as the community MIC of the target species;
(3) respectively culturing blue algae to be tested by using antibiotics with the concentration of 0, 1 and n times of the community MIC, wherein n is more than or equal to 2 and less than or equal to 8, and counting the blue algae of each group after culturing for the same number of days under the same culture condition in the step (2)The cell number of the algae is recorded as P0、P1、Pn(ii) a If Pn＜(10％～50％)P0Calculating the antibiotic heterogeneous drug resistance level f of the blue algae to be detected by the following formula:
2. the method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 1, wherein in the step (3), if f is more than or equal to 0.0001-0.00001%, the cyanobacteria to be detected has the heterogeneous drug resistance; f is less than or equal to 0.0001-0.00001%, and the blue algae to be detected has sensitivity.
3. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 1, wherein in the step (3), if P is P1≥P0And P isn≥P0The blue algae to be detected has drug resistance.
4. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 1, wherein in the step (1), a plurality of cyanobacteria target species samples are separated and purified from cyanobacteria community samples under aseptic conditions, and are respectively cultured in BG11 liquid culture medium for a plurality of days, and then are respectively concentrated until the cyanobacteria cell density is about 108-109And (2) per liter.
5. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 1, wherein in the step (2), Escherichia coli K12 for testing the activity of the antibiotic standard solution is added to a positive control group in a microdilution method.
6. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 1, wherein in the step (2), the method for judging the minimum inhibitory concentration of the antibiotics comprises the following steps: and (4) compared with a negative control group, the antibiotic concentration completely inhibits the growth of the blue algae cells and destroys the blue algae cells.
7. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotics according to claim 6, wherein the number and the integrity of the cyanobacteria cells are analyzed by optical density analysis and fluorescence microscopy.
8. The method for detecting the heterogeneous drug resistance of the cyanobacteria antibiotic according to claim 1, wherein the culture conditions in the neutralization step (3) in the step (2) are 25.0 ± 0.5 ℃, 50 ± 5 μ E/m/s and 14/10 hours light-to-dark ratio.
9. A method for detecting the stability of the heterogeneous resistance of the cyanobacteria antibiotics, which is characterized by comprising the detection method according to any one of claims 1 to 8, and further comprising the following steps:
(1) separating the blue algae with antibiotic heterogeneous drug resistance obtained by screening to obtain a plurality of blue algae colonies, and culturing the blue algae colonies for a plurality of hours by taking the antibiotic with the concentration being n times of the MIC value of the colonies;
(2) measuring the MIC value of the original cyanobacteria colony of one part of the cultured cyanobacteria colony; transferring another part of cyanobacteria colony for more than or equal to 50 times in an environment without antibiotic pressure, and then measuring the MIC value of the transferred cyanobacteria colony; and if the MIC value of the transferred cyanobacteria colony is not obviously reduced compared with the MIC value of the original cyanobacteria colony, the stability of antibiotic heterogeneous drug resistance is realized.
10. The method for detecting the stability of the heterogeneous resistance of the cyanobacteria antibiotics according to claim 9, wherein the transfer in the step (2) is performed by adding the mother liquor of the cyanobacteria colony to 50 times of the volume of the sterile BG11 liquid culture medium without the antibiotics.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210050300.5A CN114540459A (en) | 2022-01-17 | 2022-01-17 | Method for detecting heterogeneous drug resistance and stability of cyanobacteria antibiotics |
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| CN107604084A (en) * | 2017-10-30 | 2018-01-19 | 深圳市第三人民医院 | Bacterial drug resistance fast prediction system and its Forecasting Methodology |
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| WO2020061553A1 (en) * | 2018-09-21 | 2020-03-26 | Emory University | Using multiple heteroresistance to guide combination antibiotic regimens |
| CN112126682A (en) * | 2020-09-30 | 2020-12-25 | 水利部交通运输部国家能源局南京水利科学研究院 | Method for determining alternative habitat tributaries and target fishes based on environmental DNA |
| CN113862127A (en) * | 2021-09-28 | 2021-12-31 | 山东大学 | Device and method for culturing oil-producing blue algae by using sewage containing low-concentration antibiotics |
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| CN107604084A (en) * | 2017-10-30 | 2018-01-19 | 深圳市第三人民医院 | Bacterial drug resistance fast prediction system and its Forecasting Methodology |
| WO2020061553A1 (en) * | 2018-09-21 | 2020-03-26 | Emory University | Using multiple heteroresistance to guide combination antibiotic regimens |
| CN110643675A (en) * | 2019-10-31 | 2020-01-03 | 上海氘峰医疗器械有限公司 | Method for rapidly detecting drug resistance of bacteria |
| CN112126682A (en) * | 2020-09-30 | 2020-12-25 | 水利部交通运输部国家能源局南京水利科学研究院 | Method for determining alternative habitat tributaries and target fishes based on environmental DNA |
| CN113862127A (en) * | 2021-09-28 | 2021-12-31 | 山东大学 | Device and method for culturing oil-producing blue algae by using sewage containing low-concentration antibiotics |
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| ZHIYUAN WANG ET AL.: "High prevalence of unstable antibiotic heteroresistance in cyanobacteria causes resistance underestimation", 《WATER RESEARCH》 * |
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