CN201915104U - Bacteria detecting device - Google Patents
Bacteria detecting device Download PDFInfo
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- CN201915104U CN201915104U CN2010206987391U CN201020698739U CN201915104U CN 201915104 U CN201915104 U CN 201915104U CN 2010206987391 U CN2010206987391 U CN 2010206987391U CN 201020698739 U CN201020698739 U CN 201020698739U CN 201915104 U CN201915104 U CN 201915104U
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Abstract
The utility model relates to a bacteria detecting device, which comprises a filter unit, a sample injection unit and a cultivating and detecting unit. The filter unit including a hollow fibrous membrane is respectively connected with the sample injection unit and the cultivating and detecting unit and used for intercepting bacteria and/or bacteria metabolites in samples to be detected; the sample injection unit is used for injecting the samples to be detected and/or washing liquor to the inner side and/or the outer side of the hollow fibrous membrane; and the cultivating and detecting unit is used for cultivating and detecting the bacteria and/or the bacteria metabolites eluted from the filter unit. The bacteria detecting device is capable of effectively shortening detecting time and increasing detecting efficiency, and is high in detecting accuracy, long in service life and low in maintenance cost.
Description
Technical field
The utility model relates to the detection of bacterium, specifically a kind ofly carries out the device that bacterium is held back automatically and detects based on specific reaction substrate method.
Background technology
Surface water, underground water, even contain various bacteria in the rainwater.Especially when water body was subjected to human and animal excreta, sanitary sewage or the pollution of some industrial and agricultural wastewater, the bacterium in the water body can roll up.Therefore, the check to the bacteriological analysis of water, particularly intestinal bacteria has great importance on hygiology.Wherein total coli group, heat-resisting coliform and colon bacillus have been widely used as the important conventional microorganism detection index of water body safety assessment, by the detection of they and other bacterium, can judge water body hygiology quality, guarantee people's safe drinking water.
In existing water body method of detecting bacterium, generally adopt the bacterium amplification pattern that solid plate is cultivated behind liquid culture or the membrane filtration, when reaching higher level, bacterial concentration carries out artificial counting or colour developing is observed.These class methods often need fussy hand operation, comprise diluted sample, artificial inoculation, observation counting and confirmatory experiment etc., cause detecting consuming time very long, can reach 24-72 hour even more of a specified duration usually.Therefore, not only time-consuming, the effort of above-mentioned traditional water body method of detecting bacterium, and secondary pollution might take place in loaded down with trivial details operating process.
In addition, when utilizing traditional experiment chamber means that the water body in rivers, Hu Ku, swimming pool, view place is carried out Bacteria Detection, often need the large volume sampling, and to guarantee to be no more than 2 hours under the water sample haulage time normal temperature, perhaps 10 degrees centigrade of stored refrigerated are no more than 6 hours, this has brought very big inconvenience for the daily monitoring of bacterium, more can't satisfy the detection demand that people improve day by day to water environment safety.
At present, fast, the convenient and stable biochemical analysis method based on the specific reaction substrate relies on it to be easy to observe characteristics such as identification and realization automatization, become the main development trend of method of detecting bacterium in the water, obtain people's very big concern, and the existing research of adopting this method to carry out the online detection of water quality bacteria, and be used for the on-line instrument that water quality bacteria detects and come out, such as the colifast of Norway Colifast AS company and the Colilert3000 water quality colibacillus on-line monitoring instrument of France's match ring.The special indicator enzyme that this quasi-instrument utilizes the colibacillus metabolism to produce, the corresponding luminous or chromogenic substrate of hydrolysis by optical monitoring, carries out qualitative and quantitative analysis to colibacillus.Determined that initial bacterium amount is big more the detection time of this method by initial intestinal bacteria quantity, then detection time is short more.Compare with traditional method, on-line instrument can shorten detection time to 4-18 hour.In addition, this quasi-instrument can be realized the single tube continuous monitoring of water body, reduces the comprehensive detection cost, improves detection efficiency, for setting up complete water quality monitoring system and ensureing that the microorganisms in water hygienic safety provides important platform.
Can realize online detection based on the bacterium online test method of specific reaction substrate, but also have the following disadvantages the water body bacterium:
1, concentration is low, and detection time is long
Above-mentioned detection method is to cultivate from the water body direct sampling and to sample to detect, and measure relevant with the initial bacterium in the water body detection time: if the amount of bacteria in the water body is less, then detect the time that just needs cost long, detection efficiency is lower, in this case, aforesaid method just can't satisfy the rapid detection demand to water body;
2, the water body background influence is big, and accuracy in detection is low
Because above-mentioned detection method all is cultivate to detect from the water body direct sampling, water pH value, ion, free cpds or metabolite etc. can influences detected result, can't realize the accurately quantitative of bacterium in the environment water, and accuracy in detection is low.
The utility model content
In order to solve above-mentioned deficiency of the prior art, the utility model provides a kind of water quality bacteria proofing unit based on specific reaction substrate method, can deduct the water body background contamination, improve and detect initial bacterium amount, shortens detection time, improves the detected result accuracy.
For achieving the above object, the utility model adopts following technical scheme:
A kind of bacteria detecting apparatus comprises:
Filtering unit comprises hollow-fibre membrane, is connected with the cultivation detecting unit with sample injection unit respectively, is used for holding back the bacterium and/or the bacterium metabolite of sample to be tested;
Sample injection unit is used for injecting sample to be tested and/or washing fluid to the inboard and/or the outside of hollow-fibre membrane;
Cultivate detecting unit, be used to cultivate bacterium and/or bacterium metabolite, and detect from the filtering unit wash-out.
As preferably, the hollow-fibre membrane inside diameter ranges is 0～1000 μ m, and the external diameter scope is 0～1500 μ m, and membranous wall filter opening equivalence pore size filter scope is 0～1.0 μ m.
Further, described sample injection unit comprises the stream handover module, is used for optionally to filtering unit supply sample to be tested, washing fluid.
As preferably, described bacteria detecting apparatus also comprises pressure monitoring spare, links to each other the pressure of monitoring filtering unit stream with filtering unit.
The utility model also provides a kind of method of detecting bacterium, may further comprise the steps:
A, sampling step
Sample to be tested feeds the tubular fibre filtering unit, and bacterium and/or bacterium metabolite are held back by hollow-fibre membrane and accumulated in filtering unit;
Washing fluid feeds filtering unit, and the inboard and/or the outside of flushing hollow-fibre membrane develop bacterium and/or the bacterium metabolite that accumulates;
B, cultivation and detection step
Bacterium that cultivation develops and/or bacterium metabolite, the target bacteria metabolite accumulates in culturing room;
Detect the information of solution in the culturing room, obtain the information of target bacteria in the tested sample.
As preferably, described sample to be tested is a liquid sample.
As preferably, described liquid sample is water sample, liquid-food, humoral sample and beverage.
As preferably, the hollow-fibre membrane inside diameter ranges is 0～1000 μ m, and the external diameter scope is 0～1500 μ m, and membranous wall filter opening equivalence pore size filter scope is 0～1.0 μ m.
As preferably, in the described culturing room nutrient solution is arranged.
As preferably, described washing fluid is liquid nutrient medium or damping fluid or clear water.
Further, in described washing fluid and/or the nutrient solution specific reaction substrate is arranged.
As preferably, monitor the pressure of filtering unit stream.
As preferably, inboard and/or the outside and/or the sensing chamber of scavenging solution flushing hollow-fibre membrane are to Bacteria Detection stream cleaning and sterilizing.
As preferably, detect in the culturing room information of ultimate constituent in the solution or target bacteria metabolite.
As preferably, in step b, detect solution absorbency or transmittance or fluorescence intensity in the culturing room.
As preferably, in step b, whether the information of solution contains bacterium in the culturing room that obtains according to detection in the judgement sample;
Or the time dependent relation of information of solution in the culturing room that obtains according to continuous detecting, draw the concentration of bacterium.
Compared with prior art, the utlity model has following advantage:
1, effectively shortens detection time, improves detection efficiency
The employing strainer is held back the bacterium in the water body, and microbial culture is detected, and has effectively improved the initial detecting bacterium amount of bacterium, has effectively improved the water body Bacteria Detection time, especially detects for the less water body of bacterium amount, and effect is more obvious;
Owing to realized full-automatic filtration, flushing, microbial culture and the detection of water body, effectively shortened detection time, improved detection efficiency;
2, accuracy height
Adopt the hollow fibre filament of different filter openings and material, by the demand assembly and adjustment, guarantee to sample fast, efficient, but the different detection chaff interference of filtering is simultaneously got rid of the water body background interference, improves coincidence rate as a result;
Simultaneously,, have functions such as sterilization and disinfection, can not introduce secondary pollution because of the residual of manual operation or other bacterium liquid, improved detection accuracy because whole filtering unit is airtight;
3, the proofing unit life-span is long, maintenance cost is low
Because multiple rinse modes such as pressure, external pressure, interior external pressure mixing can be guaranteed effective flushing of fenestra tamper in adopting, reduce membrane damage and obstruction, make the tubular fibre filtering unit be able to permanently effective use repeatedly, reduce maintenance cost;
Adopt the stream pressure of pressure monitoring spare monitoring filtering unit, filtering unit is worked under rated pressure, tubular fibre is worked can bearing in the pressure range, prolong tubular fibre work-ing life, reduce system conversion and maintenance cost;
4, detected result stability height
Can realize the automatic assessment of filtering unit function by pressure test, guarantee repeatedly sampled result stability.
Description of drawings
Fig. 1 is a bacteria detecting apparatus structural representation among the embodiment 1;
Fig. 2 is a bacteria detecting apparatus structural representation among the embodiment 2;
Fig. 3 is a bacteria detecting apparatus structural representation among the embodiment 3;
Fig. 4 is a bacteria detecting apparatus structural representation among the embodiment 4.
Embodiment
See also Fig. 1, a kind of bacteria detecting apparatus is used for the detection of water sample bacterium; Described bacteria detecting apparatus comprises sample injection unit, filtering unit 1 and detecting unit 2;
Described sample injection unit comprises stream handover module and pipeline, and described stream handover module comprises pump and valve; Described stream handover module links to each other with filtering unit 1, sample to be tested and reagent respectively by pipeline, is used for optionally to filtering unit supply sample to be tested and reagent; In the present embodiment, reagent is washing fluid;
Described pump comprises pump 311 and pump 312, sample is driven inject filtering unit 1 inner filtration or cleaning and filtering unit 1, and described pump can be fluid drive apparatus such as peristaltic pump, surge pump, syringe pump;
Described valve comprises valve 321, valve 322, valve 325 and valve 326, is arranged on the pipeline break-make of pipeline between control sample to be tested or reagent and filtering unit 1 or filtering unit 1 and the analytical unit 2; Described valve can be two-way valve, T-valve or multiport valve etc.;
Between sample to be tested and reagent and the filtering unit 1 and filtering unit 1 and detecting unit 2 link to each other by pipeline, the pipeline internal diameter is 0.5～10mm, wall thickness is 0.1～5mm, flow range can reach 0.001～15000ml/min;
In the present embodiment, pump is peristaltic pump, and flow range is 0.07～2280ml/min, and valve is that normally closed bilateral or threeway electromagnetism breaks valve, and pipeline uses the Pharmed pipe of internal diameter 6.4mm, wall thickness 0.8mm;
Described filtering unit 1 comprises filter core and cavity, and described filter core is a hollow fiber filter assemblies, is made of hollow-fibre membrane 111, and the material of described hollow-fibre membrane 111 can be polyethylene, polysulfones, polyethersulfone, tetrafluoroethylene, polyvinylidene difluoride (PVDF) etc.; The inside diameter ranges of used hollow-fibre membrane 111 can be 0～1000 μ m, and the external diameter scope is 0～1500 μ m, and membranous wall filter opening equivalence pore size filter is 0～1.0 μ m;
In the present embodiment, described hollow fiber filter assemblies is made up of the hollow-fibre membrane of 300 hydrophilicity kynoars, and its pressure that can bear is 0.2MPa; Its internal diameter is 600 μ m, and external diameter is 800 μ m, and the equivalent pore size filter of its membranous wall is 0.1m;
Solution in the sample to be tested, metal ion, micromolecular compound and macro-molecular protein product etc. can see through the membranous wall of hollow-fibre membrane 111, bacterium or bacterium metabolite etc. then can not see through, the membranous wall that is hollow-fibre membrane 111 can play filtering effect to sample to be tested, and holds back bacterium or bacterium metabolite in the sample to be tested;
The internal space of each hollow-fibre membrane forms the filtration inner chamber jointly, and injection port 12 and outlet 13 are set at the two ends of described filter core, communicates with filtering inner chamber, and sample to be tested can flow into or the outflow filter inner chamber by injection port 12/ outlet 13;
The membranous wall of hollow-fibre membrane and cavity are formed the filtration exocoel, and outer chamber wall is provided with an exocoel opening 14, and sample to be tested can flow into or the outflow filter exocoels by exocoel opening 14;
Pump 311 links to each other with injection port 12, sample to be tested and reagent; Pump 312 links to each other with exocoel opening 14, sample to be tested and reagent, and exocoel opening 14 links to each other with waste liquid pool;
The use of filtering unit 1 has three kinds of patterns, that is: interior die pressing type, outer die pressing type and interior external pressure mixed mode: interior die pressing type is meant that pump 311 is injected sample to be tested by injection port 12; Outer die pressing type is meant that pump 312 is injected sample to be tested by exocoel opening 14; Interior external pressure mixed mode is meant that sample to be tested is injected and injected from exocoel opening 14 by pump 312 from injection port 12 by pump 311 simultaneously;
When carrying out the sample to be tested filtration, can adopt interior die pressing type, sample to be tested injects injection port 12, and then enter the filtration inner chamber, bacterium in the sample to be tested concentrates accumulation in filtering inner chamber, the membranous wall that solution, metal ion, micromolecular compound and macro-molecular protein product etc. see through hollow-fibre membrane 111 enters the filtration exocoel, and discharges filtering unit 1 by exocoel opening 14;
When carrying out the sample to be tested filtration, also can adopt outer die pressing type, be that sample to be tested injects exocoel opening 14 from valve 326, and then enter the filtration exocoel, bacterium in the sample to be tested concentrates accumulation in filtering exocoel, the membranous wall that solution, metal ion, micromolecular compound and macro-molecular protein product etc. see through hollow-fibre membrane 111 enters the filtration inner chamber, and discharges filtering unit by injection port 12/ outlet 13;
When filtering unit 1 is cleaned, can select interior die pressing type for use, scavenging solution injects injection port 12 by pump 311, and discharges from outlet 13 and exocoel opening 14; Or select outer die pressing type for use, scavenging solution injects exocoel opening 14 by pump 312, discharges from injection port 12/ outlet 13; Or external pressure mixed mode in selecting for use, scavenging solution is injected injection port 12 and is injected exocoel opening 14 by pump 312 by pump 311 simultaneously, discharges from outlet 13 discharges or other openings from the exocoel sidewall;
Described detecting unit 2 comprises cultivates module, detection module and analysis module;
Described cultivation module comprises culturing room 211 and incubator 20, and described culturing room 211 is used to hold the nutrient solution that contains the specific reaction substrate; Because present embodiment is that the total coli group in the water body is detected, the photometric signal when needing only culturing room's 211 pairs of bacterial detection luminosity information can not exert an influence; Present embodiment is to detect absorbancy or the transmittance of nutrient solution at 420nm wavelength place, and the material of selecting culturing room 211 for use is a water white transparency pyrex material, and the cutoff wavelength of this material is about 300nm, all is transparent to the light about 420nm;
Described incubator 20 is used for the envrionment conditions control of microbial culture process for can hold the casing that is used to regulate and control the microbial culture condition of described culturing room 211;
Described incubator 20 also can hold detection module simultaneously, so that detection module can detect the luminosity information of nutrient solution in the culturing room 211 in incubator 20;
Described detection module comprises light emission spare 122 and light receiving piece 123; When measuring, light emission spare 122 and light receiving piece 123 are separately positioned on the relative both sides of culturing room 211; Because present embodiment is to detect absorbancy or the transmittance of nutrient solution at 420nm wavelength place, the light that then described detection module can be launched and detect about 420nm gets final product;
Described detection module is arranged on the outside of cultivating module, and described light emission spare 122 is in the relative measurement state of light path with light receiving piece 123; When needs are measured, culturing room 211 is taken out the measurement light path of putting into detection module light emission spare 122 and light receiving piece 123 formation from incubator 20, the luminosity information of solution in the culturing room in the microbial culture process is carried out discontinuity detection;
Or detection module and analysis module 24 put into incubator 20, and make light emission spare 122 and light receiving piece 123 place the relative both sides of culturing room 211 respectively, in the microbial culture process, the luminosity information of solution in the culturing room is interrupted or continuous detecting;
In the present embodiment, detection module is provided with the outside of cultivating module;
Present embodiment also provides a kind of method of detecting bacterium, is used for the water body Bacteria Detection, may further comprise the steps:
A, provide present embodiment described bacteria detecting apparatus; Sampling specifically may further comprise the steps:
A1, filtration step
Turn on pump 311, valve 322 are closed pump 312 and all the other each valves, and the flow of control pump 311 makes the pressure of its generation can be in the tolerance range of hollow-fibre membrane 111; Pump 311 is continuously pumped into the filtration inner chamber with 2L water sample inherent filtration unit 1 injection port 12, bacterium is trapped within the hollow-fibre membrane 111 and accumulation, solution, metal ion, micromolecular compound and macro-molecular protein product etc. see through hollow-fibre membrane 111 membranous walls and enter the filtration exocoel, and enter waste liquid pool by exocoel opening 14;
A2, rinse step
Die pressing type washes bacterium in addition, turn on pump 312, valve 321 and valve 325, close pump 311 and all the other each valves, pump 312 is driven washing fluid and enters the filtration exocoel by exocoel opening 14, utilize hydraulic pressure to make washing fluid see through hollow-fibre membrane 111 membranous walls and enter the filtration inner chamber, the bacterium that will accumulate in hollow-fibre membrane 111 develops from filtering unit 1 outlet 13, brings in the culturing room 211 in the detecting unit 2; As long as washing fluid can come out the bacterium wash-out that is trapped in the hollow-fibre membrane membranous wall, can be liquid nutrient medium or damping fluid or clear water, washing fluid is a clear water in the present embodiment;
B, cultivation and detection step
B1, culturing step
With the temperature regulation to 35.5 of incubator ℃, in the sensing chamber 211 nutrient solution is arranged, contain reaction substrate ONPG (Orthonitrophenyl β-D galactopyranoside) in the nutrient solution, the target bacteria in the present embodiment is a total coli group; Growth and breeding and produce metabolite in the culturing room 211 of bacterium in detecting unit 2 comprises the excretory certain enzyme in the metabolite, described certain enzyme is decomposed the reaction substrates in the culturing room 211 specifically, discharges the feature indicator;
Along with the growing multiplication of bacterium, described feature indicator is progressively accumulation in sensing chamber 21;
If having target bacteria in the detected water sample is total coli group, then in culturing process this bacterioid with excreting beta-D tilactase (β-D galactosidase), this enzyme will cut off the ONPG molecule specifically and discharge ONP (Orhonitrophenyl) molecule, cause the ONP molecule to accumulate in culturing room 211; If not having target bacteria in the detected water sample is total coli group, then in culturing process, do not discharge the ONP molecule;
B2, detection step
Microbial culture after 10 hours, is taken out culturing room 211 in incubator, put into the light emission spare 122 of detection module and the measurement light path that light receiving piece 123 forms, the luminosity information of solution in the culturing room in the microbial culture process 211 is carried out discontinuity detection; Solution in the detection culturing room 211 is at the absorbancy or the transmittance at 420nm wavelength place, and analysis module 24 is handled the signal that light receiving pieces 123 transmit, and draws the concentration information of feature indicator ONP:
If solution absorbency reaches 0.05Unit in the final culturing room, show that then target bacteria in the detected water sample is that the concentration of total coli group is higher than 1/100mL; If absorbancy is less than 0.05Unit, show that then target bacteria in the detected water sample is that the concentration of total coli group is lower than 1/100mL.
The utility model employing tubular fibre filtering unit is held back the bacterium in the water body, then the bacterium of holding back is cultivated detection, effectively improved the initial detecting bacterium amount of bacterium, effectively improved the water body Bacteria Detection time, especially detect for the less water body of bacterium amount, effect is more obvious;
Simultaneously,, realized full-automatic filtration, flushing, microbial culture and the detection of water body, effectively shortened detection time, improved detection efficiency by sequential control to pump and valve;
Because whole filtering unit is airtight, has functions such as sterilization and disinfection, can not introduce secondary pollution because of the residual of manual operation or other bacterium liquid, has improved detection accuracy;
When the method for employing present embodiment is filtered water body, see through the discharge of hollow-fibre membrane membranous wall to detecting noisy material such as metal ion, small molecules and macro-molecular protein product etc. in the water body, effectively deduct the water body background interference, improved the accuracy of detected result.
See also Fig. 2, a kind of bacteria detecting apparatus, different with embodiment 1 described bacteria detecting apparatus is: the bacteria detecting apparatus of present embodiment also comprises pressure monitoring spare 4, is arranged on the pipeline between filtering unit and the detecting unit pressure of monitoring filtering unit stream;
Hollow fiber filter assemblies is made up of the hollow-fibre membrane 121 of 400 wetting ability polysulfones filter membrane materials, and its pressure that can bear is 0.2MPa; The internal diameter of described hollow-fibre membrane 121 is 400 μ m, and external diameter is 600 μ m, and the equivalent pore size filter of its membranous wall is 0.2 μ m;
The stream handover module also comprises valve 323 and valve 324, and described reagent also comprises scavenging solution and thimerosal; Described valve 323 links to each other with thimerosal with pump 311, pump 312 respectively, described valve 324 links to each other with scavenging solution with pump 311, pump 312 respectively, so that thimerosal and scavenging solution can enter filtering unit and detecting unit, to the cleaning that carries out disinfection of sample injection unit, filtering unit and detecting unit; Culturing room 221 links to each other with valve 327, to discharge the waste liquid in the culturing room 221; In the present embodiment, thimerosal is 0.5% clorox;
Pipeline uses the Pharmed pipe of internal diameter 3.1mm, wall thickness 1.6mm;
The light emission spare of detection module 222 can emission wavelength be the light of 365nm, and light receiving piece 223 can detect the light that wavelength is 450nm; Culturing room 221 is the colorless transparent polycarbonate material, to the emission light and the reception optical transparency of detection module;
When measuring, culturing room 221 taken out to put into from incubator 20 be arranged on the measurement light path of cultivating the detection module outside the module, solution in the culturing room in the microbial culture process is carried out discontinuity detection, or detection module and analysis module 24 put into incubator, in the process of microbial culture, the luminosity information of solution in the culturing room is interrupted or continuous detecting; In the present embodiment, detection module and analysis module 24 are arranged in the incubator.
Present embodiment also provides a kind of method of detecting bacterium, and different with embodiment 1 described detection method is:
Adopt the bacteria detecting apparatus of present embodiment;
In step a1, pump 311 is continuously pumped into the filtration inner chamber with the 1.5L water sample from injection port 12, bacterium is trapped within the hollow-fibre membrane 121 and accumulation, solution, metal ion, micromolecular compound and macro-molecular protein product etc. see through hollow-fibre membrane 121 membranous walls and enter the filtration exocoel, and enter waste liquid pool by exocoel opening 14;
In this process, the pressure of pressure monitoring spare 4 monitoring filtering unit streams, and, hollow-fibre membrane 121 is worked under rated pressure by the counter flow of transferring pump 311 of force value;
Adopt the stream pressure of pressure monitoring spare monitoring filtering unit, filtering unit is worked under rated pressure, hollow-fibre membrane 121 is worked can bearing in the pressure range, prolong tubular fibre work-ing life, reduce system conversion and maintenance cost;
In step a2,
In the present embodiment, washing fluid is for containing the liquid nutrient medium of reaction substrate MUG (4-methyl umbelliferone-β-D glucosiduronate), and target bacteria is intestinal bacteria;
With interior die pressing type flushing bacterium, turn on pump 311, valve 321 and valve 325, close pump 312 and all the other each valves, pump 311 is driven washing fluid and enters the filtration inner chamber by injection port 12, flushing hollow-fibre membrane 121 inboards, the bacterium that will accumulate in hollow-fibre membrane 121 develops from filtering unit outlet 13, brings detecting unit into;
In step b, if having target bacteria in the tested sample is intestinal bacteria, then this bacterioid is with excreting beta-D-glucuronidase in culturing process, and this enzyme will cut off the MUG molecule specifically and discharge MU (4-methyl umbelliferone) molecule, cause the MU molecule to accumulate in culturing room 21; If not having target bacteria in the detected water sample is intestinal bacteria, then in culturing process, do not discharge the MU molecule;
Simultaneously, the light emission spare of detection module 222 and light receiving piece 223 are arranged on the relative both sides of culturing room in the incubator 221, (excitation wavelength is 365nm to the fluorescence information of solution in the culturing room of detection in real time, emission wavelength is 450nm), with the concentration of obtaining feature indicator MU information over time;
When fluorescence intensity of solution in the culturing room increases and reaches preset threshold 0.01RF, cultivate and finish, record is cultivated the time that end is consumed from beginning to be cultured to, and can extrapolate colibacillary concentration in the detected water sample in conjunction with the relational expression of cultivating between elapsed time and the bacterial concentration; The relational expression of cultivating between elapsed time and the bacterial concentration can obtain by a series of water samples being pressed the contrast of preceding method and colony counting method;
The present embodiment method of detecting bacterium also comprises step c: sterilization, cleaning step:
Adopt outer die pressing type to wash, turn on pump 312, valve 323, thimerosal enters filtering unit by exocoel opening 14, continue to enter detecting unit flushing culturing room 221 from outlet 13 effusive thimerosals, after the flushing of sample to be tested volume, close each valve more than 3 times, the thimerosal storage is stayed in sample injection unit, filtering unit and the detecting unit, after reaching sterilization and wanting seeking time 4min, feed scavenging solution again and wash by same operation.
Because filtering unit is cleaned, can guarantee that the tamper of hollow-fibre membrane membranous wall filter opening is effectively washed, reduce the damage and the obstruction of hollow-fibre membrane 121, make the tubular fibre filtering unit be able to permanently effective use repeatedly, reduce maintenance cost.
Embodiment 3
See also Fig. 3, a kind of bacteria detecting apparatus, different with embodiment 2 described proofing units is: pressure monitoring spare 4 is arranged on the pipeline between sample injection unit and the filtering unit, the pressure of monitoring filtering unit stream, and present embodiment is arranged between pump 311 and the injection port 12;
Hollow fiber filter assemblies is made up of the hollow-fibre membrane 121 of 500 wetting ability polysulfones filter membrane materials;
Detection module and analysis module are arranged in the incubator; Light emission spare 322 and light receiving piece 323 are arranged on the relative both sides of culturing room in the incubator 221, can launch and detect the light of 405nm wavelength;
The present embodiment thimerosal is 0.5% DCC sodium salt, i.e. dichloro isocyanuric acid sodium.
Present embodiment also provides a kind of method of detecting bacterium, and different with embodiment 2 described detection methods is:
Provide present embodiment described bacteria detecting apparatus;
In step a, the bacterium in the excellent yogurt of 1L liquid-food accumulates in hollow-fibre membrane;
In the present embodiment, contain reaction substrate Boc-le and u-Gly-Arp-p in the washing fluid, target bacteria is the streptococcus aureus of coagulase positive;
With interior external pressure mixed mode flushing bacterium, turn on pump 311, turn on pump 312, valve 321 and valve 325 are closed all the other each valves, and washing fluid drives by injection port 12 drivings through pump 311 and enters the filtration inner chamber; Enter the filtration exocoel through pump 312 drivings by exocoel opening 14, hydraulic pressure makes washing fluid see through hollow-fibre membrane 121 membranous walls and enters the filtration inner chamber; Enter washing fluid flushing hollow-fibre membrane 121 inboards of filtering inner chamber, the bacterium that will accumulate in hollow-fibre membrane 121 develops from filtering unit outlet 13, brings detecting unit into;
In step b,, bacterium is cultivated the temperature regulation to 37 of incubator 212 ℃;
If having target bacteria in the sample is the streptococcus aureus of coagulase positive, then this bacterioid will be secreted Thrombin coagulase in culturing process, this enzyme is catalysis Boc-le and u-Gly-Arp-p reaction specifically, produce colored p-Nitroaniline (nitroa-niline, PNA), cause pna molecule in culturing room 221, to accumulate; If not having target bacteria in the detected water sample is the streptococcus aureus of coagulase positive, then in culturing process, do not discharge pna molecule;
The light emission spare of detection module 322 and light receiving piece 323 are arranged on the relative both sides of culturing room in the incubator 221, solution in the culturing room of detection in real time 221 is at the absorbancy or the transmittance at 405nm wavelength place, analysis module 24 is handled the signal that light receiving piece 323 transmits, thereby obtains the concentration information of feature indicator pna molecule; If the solution absorbance in the final culturing room then shows in the excellent yogurt of tested liquid-food to have target bacteria more than or equal to 0.1Unit; If absorbancy less than 0.1Unit, then shows in the excellent yogurt of tested liquid-food not have target bacteria;
In step c, die pressing type cleans in adopting, turn on pump 311, valve 323, thimerosal enters filtering unit by injection port 12, a thimerosal part continues to enter 221 process valves, 327 discharges of detecting unit flushing culturing room by outlet 13 through valve 325, and a part sees through to enter from hollow-fibre membrane 121 membranous walls filters exocoel from 326 discharges of exocoel opening 14 process valves; After the sample to be tested volume flushing, close each valve more than 3 times, the thimerosal storage is stayed in sample injection unit, filtering unit and the detecting unit, after reaching sterilization and wanting seeking time 5min, feed scavenging solution again and wash by same operation;
In above-mentioned Bacteria Detection step, pressure monitoring spare 4 is monitored the pressure change of filtering unit stream all the time;
The present embodiment bacteria detecting apparatus also comprises steps d: the automatic appraisal procedure of filtering unit:
The pressure change of pressure monitoring spare 4 monitoring filtering unit streams, if fully washing the back pressure change still in normal range 0.1MPa, illustrate that then cleaning, sterilization to bacteria detecting apparatus are completely, adopt this bacteria detecting apparatus can proceed to filter and detect; If pressure surpasses or is lower than initial value 0.01MPa, show may exist filter membrane to stop up or the filter membrane damage, then need continue flushing or change, until pressure change in normal range.
Can realize the automatic assessment of filtering unit function by pressure test, guarantee multiple times of filtration result's stability.
Embodiment 4
See also Fig. 4, a kind of bacteria detecting apparatus, different with embodiment 3 described bacteria detecting apparatus is: on the filtration outer chamber wall exocoel opening 15 is set again, exocoel opening 15 links to each other with valve 326, discharges the waste liquid that filters in the exocoel;
Hollow fiber filter assemblies is made up of the hollow-fibre membrane 141 of 200 wetting ability teflon materials; Its equivalent pore size filter 0.45 μ m, internal diameter 400 μ m, external diameter 600 μ m.
Present embodiment also provides a kind of method of detecting bacterium, and different with embodiment 3 described detection methods is:
Adopt the described bacteria detecting apparatus of present embodiment;
In step a, the bacterium in the 5L water body accumulates in hollow-fibre membrane 141; In the present embodiment, contain reaction substrate MUGal (4-methyl umbelliferone-β-D semi-lactosi thuja acid) in the washing fluid, target bacteria is an excrement colibacillus group;
In step b,, bacterium is cultivated the temperature regulation to 45 of incubator ℃;
If having target bacteria in the sample is excrement colibacillus group, then this bacterioid will be secreted beta-D-galactosidase in culturing process, and this enzyme will cut off the MUGal molecule specifically and discharge the MU molecule, cause the MU molecule to accumulate in culturing room; If not having target bacteria in the detected water sample is excrement colibacillus group, then in culturing process, do not discharge the MU molecule;
Simultaneously, the light emission spare of detection module 222 and light receiving piece 223 are arranged on the relative both sides of culturing room in the incubator 20 221, (excitation wavelength is 365nm to the fluorescence information of solution in the culturing room of detection in real time, emission wavelength is 450nm), with the concentration of obtaining feature indicator MU information over time;
When fluorescence intensity of solution in the culturing room increases and reaches preset threshold 0.01RF, cultivate and finish, record is cultivated the time that end is consumed from beginning to be cultured to, and can extrapolate the concentration of target bacteria in the tested sample in conjunction with the relational expression between cultivation elapsed time and the bacterial concentration; The relational expression of cultivating between elapsed time and the bacterial concentration can obtain by a series of water samples being pressed the contrast of preceding method and colony counting method;
In step c, the external pressure mixed mode cleans in adopting, turn on pump 311, pump 312, valve 323, and a thimerosal part enters filtering unit by injection port 12, and a part enters filtering unit from exocoel opening 14; Cleaned the thimerosal behind the filtering unit, a part continues to enter detecting unit flushing culturing room 221 from outlet 13, and a part is discharged from exocoel opening 15; After the sample to be tested volume flushing, close each valve more than 5 times, the thimerosal storage is stayed in sample injection unit, filtering unit and the detecting unit, after reaching sterilization and wanting seeking time 6min, feed scavenging solution again and wash by same operation.
About other explanation of the present utility model: 1, adopt hollow-fibre membrane to hold back bacterium metabolite in the sample, and bacterium metabolite developed from hollow-fibre membrane and then to the process of its detection, communicate with the listed process that detection was held back and washed to bacterium among the utility model embodiment; For the situation that has bacterium and bacterium metabolite in sample simultaneously, its processing mode also communicates, and does not repeat them here; 2, the utility model is applicable to the Bacteria Detection of liquid sample, as liquid-food, humoral sample and beverage etc., is applicable to that not only water body detects; The process of holding back, washing and detecting to corresponding liquid sample communicates with the respective process of water body bacterial monitoring; 3, detecting the information of the solution in the culturing room, both can be the information of ultimate constituent in the solution, also can be the information of bacterial indicator such as bacterium metabolite.
Above-mentioned embodiment should not be construed as the restriction to the utility model protection domain.Key of the present utility model is: realize the automatic collection and the distribution of water sample by the automatization flow path system, utilize the tubular fibre filtration module that the water body background is held back, deducted to the liquid bacterial of gathering, use flushings such as clear water, nutrient solution to concentrate thalline, under controlled condition, carry out the bacterium amplification after flowing into the cultivation detecting unit, the signal that monitoring specific reaction substrate produces, according to the particular kind of relationship of signal value and bacterial concentration, realize automatic, the online quantitative or early warning analysis of water body bacterium.Under the situation that does not break away from the utility model spirit, any type of change that the utility model is made all should fall within the protection domain of the present utility model.
Claims (4)
1. bacteria detecting apparatus comprises:
Filtering unit comprises hollow-fibre membrane, is connected with the cultivation detecting unit with sample injection unit respectively, is used for holding back the bacterium and/or the bacterium metabolite of sample to be tested;
Sample injection unit is used for injecting sample to be tested and/or washing fluid to the inboard and/or the outside of hollow-fibre membrane;
Cultivate detecting unit, be used to cultivate bacterium and/or bacterium metabolite, and detect from the filtering unit wash-out.
2. bacteria detecting apparatus according to claim 1 is characterized in that: the hollow-fibre membrane inside diameter ranges is 0～1000 μ m, and the external diameter scope is 0～1500 μ m, and membranous wall filter opening equivalence pore size filter scope is 0～1.0 μ m.
3. bacteria detecting apparatus according to claim 1 is characterized in that: described sample injection unit comprises the stream handover module, is used for optionally to filtering unit supply sample to be tested, washing fluid.
4. according to the described bacteria detecting apparatus of the arbitrary claim of claim 1～3, it is characterized in that: described bacteria detecting apparatus also comprises pressure monitoring spare, links to each other the pressure of monitoring filtering unit stream with filtering unit.
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| CN2010206987391U CN201915104U (en) | 2010-12-31 | 2010-12-31 | Bacteria detecting device |
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| CN2010206987391U CN201915104U (en) | 2010-12-31 | 2010-12-31 | Bacteria detecting device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175632A (en) * | 2010-12-31 | 2011-09-07 | 聚光科技(杭州)股份有限公司 | Method and device for detecting bacteria |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102175632A (en) * | 2010-12-31 | 2011-09-07 | 聚光科技(杭州)股份有限公司 | Method and device for detecting bacteria |
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Effective date of registration: 20120323 Address after: Hangzhou City, Zhejiang province Binjiang District 310052 shore road 760 Co-patentee after: Wuxi Juguang Shengshi Sensor Network Co., Ltd. Patentee after: Focused Photonics (Hangzhou) Inc. Co-patentee after: Hangzhou Juguang Environmental Protection Technology Co.,Ltd. Address before: Hangzhou City, Zhejiang province Binjiang District 310052 shore road 760 Co-patentee before: Wuxi Juguang Shengshi Sensor Network Co., Ltd. Patentee before: Focused Photonics (Hangzhou) Inc. |
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