CN105543078A - Body fluid enrichment culture bottle and preparation method thereof - Google Patents
Body fluid enrichment culture bottle and preparation method thereof Download PDFInfo
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- CN105543078A CN105543078A CN201610090747.XA CN201610090747A CN105543078A CN 105543078 A CN105543078 A CN 105543078A CN 201610090747 A CN201610090747 A CN 201610090747A CN 105543078 A CN105543078 A CN 105543078A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/38—Caps; Covers; Plugs; Pouring means
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/06—Nozzles; Sprayers; Spargers; Diffusers
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2296—Oxides; Hydroxides of metals of zinc
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Abstract
The invention discloses a body fluid enrichment culture bottle and a preparation method thereof. The bottle comprises a bottle body, a bottle plug, a bottle cap a sensor and a fluid medium. The bottle body is in a cylindrical shape. The sensor is attached to the bottom inside the bottle body. The bottle plug and the bottle cap achieve the sealing and mechanical protecting effects. The sealed space of the body fluid enrichment culture bottle is the negative pressure aseptic environment. The bottle body of the body fluid enrichment culture bottle is the dual-layer plastic bottle body, one layer is made of plastic materials with strength support, and the other layer is made of plastic materials with gas barrier effect. The two layers of plastic materials avoid breaking safety risk and breathability influences. Macromolecule silicone rubber materials in the sensor are solidified at high temperature, so that the preparation time and solidification time of the sensor are greatly shortened, and the time of the whole production process is shortened. The sensor is nontransparent due to an opacifying agent, light signal collection is not interfered, and then the sensitivity of automatic detection equipment is improved.
Description
Technical field
The present invention relates to Clinical microorganism detection technique field, specifically, is a kind of body fluid enriched culture bottle and preparation method thereof.
Background technology
Body fluid cultivate be in human body sample with or without microorganism exist a kind of microbiological test method, comprise hemoculture, Sputum culturing, urine cultivate and other body fluid (hydrothorax, ascites, cerebrospinal fluid etc.) cultivate.Rapid detection seriously being jeopardized clinically to septicemia, the microbemia of patient vitals, whether have microorganism growth with very important effect of clarifying a diagnosis in patient body fluid, is the key of clinical effective treatment.So the detection of pathogenic microorganism should be carried out within the time short as far as possible in body fluid or analogue.In recent years, along with scientific-technical progress, the combination of microbiology, computer and engineering science, has createed the culture systems of many automatizations.
The body fluid microorganism culturing of automatization generally comprises two portions, automatic detection system and body fluid enriched culture bottle.Automatic detection system provides incubator and the detection system of specific culture environment, and body fluid enrichment medium provides microbial growth environment and the sensor for detection.Body fluid sample is inoculated in body fluid enriched culture bottle, cultivates in instrument at automatization body fluid and carry out microorganism culturing, the change that detection system is produced by sensor in body fluid enriched culture bottle, automatically detect, provide cultivation results, be supplied to the foundation of clinical diagnosis.
Existing culturing bottle mostly is flashing lightning magnetic field detector, in the patent No. 2004200480078, a kind of flashing lightning magnetic field detector is described, this bottle does not have negative pressure, be not easy to the injection of clinical sample, can pollute when using careless, and use glass body, there is security risk, can fragmentation be caused when accidentally falling, make user be exposed to glass cullet, and have the possibility of infection.Another kind of use plastic material, such as polycarbonate, the bottle of this material can solve security risk, but it has ventilation property, can cause the negative pressure variation in culturing bottle, can not ensure smooth injecting sample between effective service life.If ambient atmos enters in culturing bottle on the other hand, built-in gaseous constituent in culturing bottle can be caused to change, and affect microbial growth, after such as, entering air in Anaerobic culturel bottle, anerobe can be caused to grow, and cause clinical erroneous judgement.
Sensor bottom culturing bottle is made up of high molecular polymer and indicator, in prior art, the perviousness of sensor is determined by molecular size, as described in the patent No. 2012105834227, polymer matrix can stop the material of molecular weight more than 300, small molecules is there is, water, hydrogen ion etc., if these small molecules all can have an impact to sensor through semi-permeable membranes in real liquid substratum, cause colour-change, and essence not the change that microorganism growth produces.And according to sensor described in the prior, there is light transmission, and the Cleaning Principle that automatic detection system uses is optical detection principle, gathers optical signal, be then converted into electric signal transmission in Controlling System, if now sensor has light transmission, the collection of optical signal will be affected.
In addition, in the prior art, the preparation of sensor and the time spent by solidification all longer, generally at 24 hours-48 hours, had a strong impact on production efficiency.
Summary of the invention
Technical problem to be solved by this invention is, body fluid enriched culture bottle providing a kind of plastic material of bilayer and preparation method thereof, not easily broken, and gas tightness is transparent, Heat stability is good, and manufacturing process is simple, reduces production process step, reduces production cost.
In order to solve the problems of the technologies described above; the technical solution used in the present invention is: a kind of body fluid enriched culture bottle; comprise bottle, bottle stopper, bottle cap, sensor, liquid nutrient medium; culturing bottle bottle is cylindrical; interior bottom portion at the bottom of sensor is affixed on bottle; bottle stopper and bottle cap play sealing and mechanical protection effect; the seal cavity of body fluid enriched culture bottle is the gnotobasis of negative pressure; the bottle of body fluid enriched culture bottle is double-layer plastic bottle; one deck is the plastic material with strength support, and another layer is for having the plastic material of gas shield effect.
The described plastic material with strength support is nylon, polycarbonate or polypropylene PP, the skin of described double-deck bottle or internal layer are the plastic material with gas shield effect, described in there is gas shield effect plastic material be PEN, polyethylene terephthalate, polyester or polyvinyl chloride.
The sensor of described body fluid enriched culture bottle is selectivity semi-permeable membranes, and it can stop polar molecule through non-polar molecule, ensures that carbonic acid gas can stop other molecules in substratum and/or ion through semi-permeable membranes.
The sensor of described body fluid enriched culture bottle is polymer silastic material polymethyl siloxane and solidifying agent thereof.
The sensor of described body fluid enriched culture bottle contains opalizer, acid base indicator and fluorescent indicator, can there is colour-change according to the carbonic acid gas of microorganism growth generation in substratum in acid base indicator, fluorescent indicator indicator according to the carbonic acid gas of microorganism growth generation in substratum, fluorescent reaction can occur.
Described acid base indicator is dibromothymolsulfonphthalein, purpurum bromocresolis, tetrabromo-mcresolsulfonphthalein or xylenol blue; Described fluorescent indicator is fluorescein, pararosaniline, Sulforhodamine B, rhodamine B, C.I. 42685, sodium alizarinsulfonate or Xylene Red; Described opalizer is zinc white, titanium white, lithopone or titanox.
Described sensory device material formula is:
Above per-cent is volume percent, wherein acid base indicator is the aqueous solution of mass percent concentration 0.1%-10%, opalizer be mass percent concentration 10%-80% be suspended the aqueous solution, fluorescent indicator is the aqueous solution of mass percent concentration 0.1-10%, and other compositions are water and/or ethanol.
Preferably, described sensory device material formula is:
Above per-cent is volume percent, wherein dibromothymolsulfonphthalein is the dibromothymolsulfonphthalein aqueous solution of mass percent concentration 0.1%-10%, zinc white is that the zinc white of mass percent concentration 10%-80% is suspended the aqueous solution, rhodamine B is the rhodamine B aqueous solution of mass percent concentration 0.1-10%, and other compositions are water and/or ethanol.
Described liquid nutrient medium comprises tryptic soy broth, brain heart infusion, yeast extract, carbohydrate, and containing microorganism being played to the vitamin B complex of repair and/or vitamin K race and/or vitamin-E race.
Described liquid nutrient medium also comprises the Sodium Citrate, sodium thioglycollate, Trisodium Citrate, ironic citrate, S-WAT, one or more in L-cysteine hydrochloride that promote anaerobion growth; Comprise and reduce other materials interference in sample, one or more in the edetate of accelerate growth of microorganism, para-amino benzoic acid, poly-fennel sodium sulfonate, glucose oxidase, protohemine.
Be added with the polymeric adsorbent of mass percent 2%-20% in described liquid nutrient medium, polymeric adsorbent is one or more non-ionic adsorption resins and Zeo-karb, to weaken the interference that in sample, microbiotic is cultivated sample.
The inside enclosed space of described body fluid enriched culture bottle contains built-in gas, and described built-in gas comprises one or more gases in oxygen, nitrogen, carbonic acid gas, hydrogen, air.
A preparation method for body fluid enriched culture bottle, comprises the following steps:
(1) preparation of Double layer culture bottle: first form double-deck parison, then carry out blowing, the temperature of blowing is at 25 degrees Celsius to 150 degrees Celsius;
(2) sensor material preparation: fluorescent indicator and acid base indicator purified water are dissolved, opalizer purified water is mixed with and is suspended the aqueous solution, proportionally after mixing, add the silastic material containing platinum group catalyst, solidifying agent, stir deaeration under vacuum, while stirring, complete deaeration work;
(3) sensor adds and solidification: use even dispensing apparatus to be incorporated in bottle by sensor material, pave in bottom, forms the sensor of 2-10mm thickness; The bottle introducing sensor material is solidified under temperature 45-85 DEG C of environment, in 1-2 hour, completes solidification;
(4) medium preparing: liquid nutrient medium component added in proportion in purified water, dissolves, stirs, degerming;
(5) filling: by the liquid nutrient medium of sterilizing and the polymeric adsorbent of sterilizing by filling-to jump a queue-sealing machine is filled in the bottle containing sensor;
(6) seal: the plug of sterilizing is attached to above bottleneck, by filling--sealing machine blanketing gas of jumping a queue, and forms vacuum, then by plug compacting, finally by filling-to jump a queue-aluminium-plastic cap is attached to above bottleneck by sealing machine, then compacting;
(7) sterilizing: good seal, culturing bottle that each assembly apparatus is good are carried out irradiation sterilization, the final aseptic body fluid Zengjing Granule bottle product forming sealing.
The invention has the beneficial effects as follows:
1, a kind of culturing bottle bottle of plastic material of bilayer is provided, solves risk, the complex process of breakable security risk, the built-in gaseous constituent of ventilative impact and internal negative pressure, the risk that cost is high simultaneously.
2, a kind of sensor based on acid base indicator and fluorescent indicator is provided, realize the detection method of sensor colors change and fluorescent reaction simultaneously, solve the risk relying on acid base indicator colour-change easily to produce false positive result, can not artificial interpretation be carried out with fluorescent indicator and the high shortcoming of automatic checkout equipment cost, improve sensitivity and the accuracy rate of whole detection system.
The sensor of the body fluid enriched culture bottle 3, used is selectivity semi-permeable membranes, it can stop polar molecule through non-polar molecule, ensure that carbonic acid gas can stop other molecules in substratum and/or ion through semi-permeable membranes, the concentration of the carbonic acid gas that the colour-change of sensor is only produced by microorganism growth in sample causes, and exempt the interference of other ions and/or molecule, and then improve the accuracy rate of equipment Inspection.
4, the polymer silastic material in sensor used comprises polymethyl siloxane, solidifying agent, platinum group catalyst, and solidifies under the high temperature conditions, greatly can shorten preparation time and the set time of sensor, and then shorten the time of whole production process.
5, the opalizer in sensor used comprises zinc white, titanium white, lithopone, titanox, can play interception, makes sensor opaque, gathers optical signal and is not interfered, and then improve the sensitivity of automatic detection system.
Add in the liquid nutrient medium of 6, described body fluid enriched culture bottle and the vitamin B complex of repair and/or vitamin K race and/or vitamin-E race are risen to microorganism, the microorganism cells (containing microbiotic, the microorganism that may exist being sustained damage in such as blood sample) sustained damage in sample can be made to be repaired, increase and detect probability, avoid clinical undetected and incur loss through delay conditions of patients.
The Sodium Citrate, sodium thioglycollate, Trisodium Citrate, ironic citrate, S-WAT, one or more in L-cysteine hydrochloride that promote anaerobion growth are added in the liquid nutrient medium of 7, described body fluid enriched culture bottle, to the oxidizing substance neutralization introduced in liquid nutrient medium or in sample, ensure the anaerobic environment of anaerobion growth.What increase the especially obligatory anaerobic bacteria of anerobe detects probability, avoids clinical undetected and incur loss through delay conditions of patients.
8, provide a kind of simple manufacturing process, adopt final irradiation sterilization, reduce the clean requirement to environment in production process, reduce production process step, reduce production cost.
Accompanying drawing explanation
Fig. 1 body fluid enriched culture bottle structural representation.
The production process of Fig. 2 body fluid enriched culture bottle.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail:
As illustrated in fig. 1 and 2, body fluid enriched culture bottle of the present invention, comprises bottle 1, bottle stopper 3, bottle cap 2, sensor 6, liquid nutrient medium 5; Wherein said culturing bottle bottle is cylindrical, interior bottom portion at the bottom of sensor 6 is affixed on bottle, and bottle stopper 3 and bottle cap 2 play sealing and mechanical protection effect, and culturing bottle inside is gnotobasis.
The bottle 1 of described body fluid enriched culture bottle is double-layer plastic bottle, the internal layer of described double-deck bottle or skin are for having the plastic material of strength support, such as nylon, polycarbonate or polypropylene PP, the skin of described double-deck bottle or internal layer are the plastic material with gas shield effect, such as PEN, polyethylene terephthalate, polyester or polyvinyl chloride.
The seal cavity of described body fluid enriched culture bottle is negative pressure;
The bottle stopper 3 of described body fluid enriched culture bottle is elastic butyl rubber plug, fits tightly, play sealing function with bottleneck;
The bottle cap 2 of described body fluid enriched culture bottle is aluminium-plastic cap, and to guarantee that bottle stopper can not drop and pollute, during use, the plastic cover 8 on top layer can remove, convenient operation.
The sensor 6 of described body fluid enriched culture bottle is selectivity semi-permeable membranes, and it can stop polar molecule through non-polar molecule, ensures that carbonic acid gas can stop other molecules in substratum and/or ion through semi-permeable membranes.
The sensor of described body fluid enriched culture bottle uses polymer silastic material.
The sensor of described body fluid enriched culture bottle contains opalizer.
The sensor of described body fluid enriched culture bottle contains acid base indicator, and this indicator according to the carbonic acid gas of microorganism growth generation in substratum, colour-change can occur.
Described acid base indicator comprises dibromothymolsulfonphthalein, purpurum bromocresolis, tetrabromo-mcresolsulfonphthalein, xylenol blue etc.
The sensor of described body fluid enriched culture bottle contains fluorescent indicator, and this indicator according to the carbonic acid gas of microorganism growth generation in substratum, fluorescent reaction can occur.
Described fluorescent indicator comprises fluorescein, pararosaniline, Sulforhodamine B, rhodamine B, C.I. 42685, sodium alizarinsulfonate, Xylene Red etc.
Polymer silastic material in described sensor comprises polymethyl siloxane.
Opalizer in described sensor comprises zinc white, titanium white, lithopone, titanox.
The liquid nutrient medium of described body fluid enriched culture bottle is the nutritive ingredient such as tryptic soy broth, brain heart infusion, yeast extract, carbohydrate that most Institute of Micro-biology needs.
The liquid nutrient medium of described body fluid enriched culture bottle also comprises and plays the vitamin B complex of repair and/or vitamin K race and/or vitamin-E race to microorganism.
The liquid nutrient medium of described body fluid enriched culture bottle also comprises the Sodium Citrate, sodium thioglycollate, Trisodium Citrate, ironic citrate, S-WAT, one or more in L-cysteine hydrochloride that promote anaerobion growth.
The liquid nutrient medium of described body fluid enriched culture bottle also comprises and reduces other materials interference in sample, one or more in the edetate of accelerate growth of microorganism, para-amino benzoic acid, poly-fennel sodium sulfonate, glucose oxidase, protohemine.
The polymeric adsorbent of 2%-20% (w/w) is added with, to weaken the interference that in sample, microbiotic is cultivated sample in the liquid nutrient medium of described body fluid enriched culture bottle.
The inside enclosed space of described body fluid enriched culture bottle contains built-in gas, and described built-in gas comprises one or more gases in oxygen, nitrogen, carbonic acid gas, hydrogen, air.
The preparation method of body fluid enriched culture bottle of the present invention.Have monolayer plastic culturing bottle or glass culturing bottle in prior art, mentioned the defect that it exists above, also have a kind of laminated plastics culturing bottle, it can meet not easily broken, the features such as gas tightness, but complex process, and preparation difficulty, cost is higher.
Therefore, the invention provides a kind of not easily fragmentation that can meet required for culturing bottle, gas tightness, transparent, the body fluid enriched culture bottle that thermostability etc. need.Final irradiation sterilization, finally forms sterile product, and the sterility requirements reduced in process and sterilization steps.
The production process of described body fluid enriched culture bottle comprises, the preparation of double-layer plastic bottle, the preparation of sensor material, the interpolation of sensor and solidification, and the preparation of liquid nutrient medium is filling, sealing, sterilizing.
The preparation process of described Double layer culture bottle comprises: form parison; Blow molding.
Described sensor material preparation process comprises: fluorescent indicator and acid base indicator purified water are dissolved, opalizer purified water is mixed with and is suspended the aqueous solution, is proportionally mixed in silicon rubber after mixing.
Described sensor adds and solidification process comprises: use even dispensing apparatus to be incorporated into by sensor material in culturing bottle bottle, through solidification in 1-2 hour under 45-85 DEG C of condition.
Described sensor solidification process comprises use catalyzer and solidifying agent, and described catalyzer is the platinum group catalyst added in sensor components.
Described liquid nutrient medium preparation process comprises: substratum moiety added in proportion in purified water, fully dissolve, through sterilising treatment.
Described pouring process comprises: the polymeric adsorbent equivalent after sterilizing be dispensed into and prepare in the culturing bottle of sensor.
Described pouring process also comprises: the liquid nutrient medium after filtration sterilization is added to preparing in the culturing bottle of sensor.
Described seal process comprises: above the culturing bottle butyl rubber plug after sterilizing being placed in filling good polymeric adsorbent and liquid nutrient medium, by filling-to jump a queue-sealing machine is to culturing bottle filled with gas.
Described seal process also comprises: above the culturing bottle butyl rubber plug after sterilizing being placed in filling good polymeric adsorbent and liquid nutrient medium, by filling-to jump a queue-sealing machine applies vacuum to culturing bottle inside, form negative pressure.
Described seal process also comprises: be pressed in by aluminium-plastic cap and injected the culturing bottle bottleneck that filled with gas forms negative pressure, compacting seals.
Described sterilization process is irradiation sterilization.
Body fluid enriched culture bottle of the present invention, can not only carry out blood cultivation, can also cultivate, as hydrothorax, ascites, cerebrospinal fluid etc. other body fluid samples clinical.
Specifically, described culturing bottle bottle is double-layer plastic bottle 1, the air-locked cylinder type of water white transparency, and inside is sterile negative pressure environment, and sample is injected in culturing bottle smoothly, exempts Pollution risk.Bottle stopper 3 is elasticity butyl rubber plug, fits tightly with bottleneck the resistance to air loss ensureing culturing bottle.Bottle cap 2 is double-deck aluminium-plastic cap, is removed by the plastic cap 8 on top layer during use, uses front plastic cap will to keep the sealing of plug and aseptic.In culturing bottle, the liquid nutrient medium 4 of configuration is the nutrient media components such as tryptic soy broth, brain heart infusion, yeast extract, carbohydrate that most microorganism growth needs, and the VITAMIN of adding suitably to its repair of microorganism cells, make microorganism Fast-propagation, and the microorganism being subject to interfering substance in sample (such as microbiotic) and damaging can be repaired, improve and detect probability.Be added with the material of antagonistic action in liquid nutrient medium, as one or more in edetate, para-amino benzoic acid, poly-fennel sodium sulfonate, glucose oxidase, protohemine, microbial growth and Fast-propagation can be promoted.Polymeric adsorbent 5 is added with in liquid nutrient medium, polymeric adsorbent is one or more non-ionic adsorption resins and Zeo-karb, polymeric adsorbent is suspended in liquid nutrient medium, for adsorbing the materials such as the microbiotic that contains in sample, reduce the interference that the materials such as microbiotic are cultivated body fluid, improve recall rate.Containing built-in gas in body fluid enriched culture bottle, provide the prerequisite of microorganism growth.At the bottom of the bottle of body fluid enriched culture bottle, inject sensor 6, sensor is selectivity semi-permeable membranes, adopt the preparation of polymer silastic material, it can stop polar molecule through non-polar molecule, ensures that carbonic acid gas can stop other molecules in substratum and/or ion through semi-permeable membranes, and the concentration of the carbonic acid gas that the colour-change of sensor is only produced by microorganism growth in sample causes, and exempt the interference of other ions and/or molecule, and then improve the accuracy rate of equipment Inspection.Containing catalyzer in sensor, sensor material can be made to introduce fast setting after in culturing bottle, shorten set time.Containing acid base indicator and fluorescent indicator in sensor, when microorganism growth in bottle produces carbonic acid gas, cause acid base indicator colour-change and the change of fluorescent indicator fluorescent substance.Contain built-in blanketing gas 7 in body fluid enriched culture bottle, comprise one or more in nitrogen, oxygen, hydrogen, carbonic acid gas, air, meet inhomogeneity microbial growth demand, promote microbial growth.
As shown in Figure 2, the production process of body fluid enriched culture bottle of the present invention, needs 7 steps, specifically describes as follows:
Prepared by step 1 double-layer plastic bottle: first use parison mould casting parison internal layer, under 120-150 DEG C of condition, polycarbonate is injected mould, after cooling, the inner core casting parison of the outer mould of parison is outer according to this, under 70-100 DEG C of condition, PEN is injected parison mould, mould adobe blocks after cooling, then double bottle mould is proceeded to, under the high temperature conditions blow molding.
Prepared by step 2 sensor material: take rhodamine B 0.0496g, be dissolved in, in 10mL purified water, take dibromothymolsulfonphthalein 0.345g, be dissolved in 15mL purified water, taking zinc white 3.472g joins in 10mL purified water, solution will be prepared above, mix with 495.9mL silicon rubber basal component and 49.6mL solidifying agent, with purified water constant volume to 600mL, stir deaeration 15min under vacuum, while stirring, complete deaeration work.
Step 3 sensor adds: use even dispensing apparatus to be incorporated in bottle by sensor material, pave in bottom, forms the sensor of 2-10mm thickness.
Step 4 sensor solidifies: solidified under temperature 45-85 DEG C of environment by the bottle introducing sensor material, the solidifying agent contained in silicon rubber and platinum group catalyst play promotion solidification, in 1-2 hour, complete solidification, and sensor preparation work completes.
Prepared by step 5 liquid nutrient medium: take tryptic soy broth 90g, brain heart infusion 6g, yeast extract 6g, glucose 30g, sodium-chlor 15g, poly-fennel sodium sulfonate 0.03g, combination VITAMIN 0.03g, protohemine 0.003g, somatomedin 0.003g, adds in 3000mL purified water by above liquid nutrient medium component, dissolves, stir, filtration sterilization.
Step 6 is filling: by the liquid nutrient medium of sterilizing and the polymeric adsorbent of sterilizing by filling-to jump a queue-sealing machine is filled in the bottle containing sensor, every bottle of 30mL.
Step 7 seals: be attached to above bottleneck by the plug of sterilizing, by filling--sealing machine blanketing gas of jumping a queue, and forms vacuum, then by plug compacting, finally by filling-to jump a queue-aluminium-plastic cap is attached to above bottleneck by sealing machine, then compacting.
Step 8 sterilizing: will with good seal, the culturing bottle that each assembly apparatus is good carries out irradiation sterilization (25kGy), the final aseptic body fluid Zengjing Granule bottle product forming sealing.
Conducted further description by some embodiments with regard to some committed steps below.
In one embodiment, the material of sensor comprises:
Wherein Sylgard184 silicon rubber basal component comprises polymethyl siloxane and platinum group catalyst.
In yet another embodiment, the material of sensor comprises:
In an embodiment, the material of sensor comprises:
In another embodiment, the material of sensor comprises:
The scope of sensory device material formula in the present invention:
Above-described is volume ratio, wherein dibromothymolsulfonphthalein is the aqueous solution of concentration 0.1%-10% (w/w), zinc white is the suspension liquid of concentration 10%-80% (w/w), rhodamine B is the aqueous solution of concentration 0.1-10% (w/w), and other compositions are water or ethanol etc.
In one embodiment, the preparation method of liquid nutrient medium is taken in proportion by following component, is then dissolved in purified water, and heating makes it fully dissolve.
In another embodiment, liquid nutrient medium comprises:
Above-described embodiment is only for illustration of technological thought of the present invention and feature, its object is to enable those skilled in the art understand content of the present invention and implement according to this, only can not limit the scope of the claims of the present invention with the present embodiment, namely the equal change done of all disclosed spirit or modification, still drop in the scope of the claims of the present invention.
Claims (13)
1. a body fluid enriched culture bottle; comprise bottle (1), bottle stopper (3), bottle cap (2), sensor (5), liquid nutrient medium (4); culturing bottle bottle is cylindrical; interior bottom portion at the bottom of sensor is affixed on bottle; bottle stopper and bottle cap play sealing and mechanical protection effect; the seal cavity of body fluid enriched culture bottle is the gnotobasis of negative pressure; it is characterized in that; the bottle (1) of body fluid enriched culture bottle is double-layer plastic bottle; one deck is the plastic material with strength support, and another layer is for having the plastic material of gas shield effect.
2. body fluid enriched culture bottle according to claim 1, it is characterized in that, the described plastic material with strength support is nylon, polycarbonate or polypropylene PP, the skin of described double-deck bottle or internal layer are the plastic material with gas shield effect, described in there is gas shield effect plastic material be PEN, polyethylene terephthalate, polyester or polyvinyl chloride.
3. body fluid enriched culture bottle according to claim 1, it is characterized in that, the sensor of described body fluid enriched culture bottle is selectivity semi-permeable membranes, it can stop polar molecule through non-polar molecule, ensures that carbonic acid gas can stop other molecules in substratum and/or ion through semi-permeable membranes.
4. body fluid enriched culture bottle according to claim 3, is characterized in that, the sensor of described body fluid enriched culture bottle is polymer silastic material polymethyl siloxane and solidifying agent thereof.
5. body fluid enriched culture bottle according to claim 1, it is characterized in that, the sensor of described body fluid enriched culture bottle contains opalizer, acid base indicator and fluorescent indicator, can there is colour-change according to the carbonic acid gas of microorganism growth generation in substratum in acid base indicator, fluorescent indicator indicator according to the carbonic acid gas of microorganism growth generation in substratum, fluorescent reaction can occur.
6. body fluid enriched culture bottle according to claim 5, is characterized in that, described acid base indicator is dibromothymolsulfonphthalein, purpurum bromocresolis, tetrabromo-mcresolsulfonphthalein or xylenol blue; Described fluorescent indicator is fluorescein, pararosaniline, Sulforhodamine B, rhodamine B, C.I. 42685, sodium alizarinsulfonate or Xylene Red; Described opalizer is zinc white, titanium white, lithopone or titanox.
7. the body fluid enriched culture bottle according to any one of claim 4-6, is characterized in that, described sensory device material formula is:
Above per-cent is volume percent, wherein acid base indicator is the aqueous solution of mass percent concentration 0.1%-10%, opalizer be mass percent concentration 10%-80% be suspended the aqueous solution, fluorescent indicator is the aqueous solution of mass percent concentration 0.1-10%, and other compositions are water and/or ethanol.
8. body fluid enriched culture bottle according to claim 7, is characterized in that, described sensory device material formula is:
Above per-cent is volume percent, wherein dibromothymolsulfonphthalein is the dibromothymolsulfonphthalein aqueous solution of mass percent concentration 0.1%-10%, zinc white is that the zinc white of mass percent concentration 10%-80% is suspended the aqueous solution, rhodamine B is the rhodamine B aqueous solution of mass percent concentration 0.1-10%, and other compositions are water and/or ethanol.
9. body fluid enriched culture bottle according to claim 1, it is characterized in that, described liquid nutrient medium comprises tryptic soy broth, brain heart infusion, yeast extract, carbohydrate, and containing microorganism being played to the vitamin B complex of repair and/or vitamin K race and/or vitamin-E race.
10. body fluid enriched culture bottle according to claim 9, it is characterized in that, described liquid nutrient medium also comprises the Sodium Citrate, sodium thioglycollate, Trisodium Citrate, ironic citrate, S-WAT, one or more in L-cysteine hydrochloride that promote anaerobion growth; Comprise and reduce other materials interference in sample, one or more in the edetate of accelerate growth of microorganism, para-amino benzoic acid, poly-fennel sodium sulfonate, glucose oxidase, protohemine.
11. body fluid enriched culture bottles according to claim 10, it is characterized in that, the polymeric adsorbent of mass percent 2%-20% is added with in described liquid nutrient medium, polymeric adsorbent is one or more non-ionic adsorption resins and Zeo-karb, to weaken the interference that in sample, microbiotic is cultivated sample.
12. body fluid enriched culture bottles according to claim 1, it is characterized in that, the inside enclosed space of described body fluid enriched culture bottle contains built-in gas, and described built-in gas comprises one or more gases in oxygen, nitrogen, carbonic acid gas, hydrogen, air.
The preparation method of 13. 1 kinds of body fluid enriched culture bottles, is characterized in that, comprises the following steps:
(1) preparation of Double layer culture bottle: first form double-deck parison, then carry out blowing, the temperature of blowing is at 25 degrees Celsius to 150 degrees Celsius;
(2) sensor material preparation: fluorescent indicator and acid base indicator purified water are dissolved, opalizer purified water is mixed with and is suspended the aqueous solution, proportionally after mixing, add the silastic material containing platinum group catalyst, solidifying agent, stir deaeration under vacuum, while stirring, complete deaeration work;
(3) sensor adds and solidification: use even dispensing apparatus to be incorporated in bottle by sensor material, pave in bottom, forms the sensor of 2-10mm thickness; The bottle introducing sensor material is solidified under temperature 45-85 DEG C of environment, in 1-2 hour, completes solidification;
(4) medium preparing: liquid nutrient medium component added in proportion in purified water, dissolves, stirs, degerming;
(5) filling: by the liquid nutrient medium of sterilizing and the polymeric adsorbent of sterilizing by filling-to jump a queue-sealing machine is filled in the bottle containing sensor;
(6) seal: the plug of sterilizing is attached to above bottleneck, by filling--sealing machine blanketing gas of jumping a queue, and forms vacuum, then by plug compacting, finally by filling-to jump a queue-aluminium-plastic cap is attached to above bottleneck by sealing machine, then compacting;
(7) sterilizing: good seal, culturing bottle that each assembly apparatus is good are carried out irradiation sterilization, the final aseptic body fluid Zengjing Granule bottle product forming sealing.
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| CN201610090747.XA CN105543078A (en) | 2016-02-18 | 2016-02-18 | Body fluid enrichment culture bottle and preparation method thereof |
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| CN201610090747.XA CN105543078A (en) | 2016-02-18 | 2016-02-18 | Body fluid enrichment culture bottle and preparation method thereof |
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