CN101701184A - Microbe rapid detection device and detection method thereof - Google Patents
Microbe rapid detection device and detection method thereof Download PDFInfo
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- CN101701184A CN101701184A CN200910309576A CN200910309576A CN101701184A CN 101701184 A CN101701184 A CN 101701184A CN 200910309576 A CN200910309576 A CN 200910309576A CN 200910309576 A CN200910309576 A CN 200910309576A CN 101701184 A CN101701184 A CN 101701184A
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Abstract
The invention provides a microbe rapid detection device and a detection method thereof, and the device comprises a detection box, a sample bottle and a reference bottle in the box, wherein the sample bottle and the reference bottle are internally provided with a sample to be detected and a sterile reference object respectively; the detection box is a closed box body connected with a heating device and a vacuum-pumping device and is integrated by nesting an inner box, a heat-insulation interlayer and an outer box into a whole; the top surface of the inner box is distributed with bottle-plugging holes; the sample bottle and the reference bottle are plugged into the inner box, and the outer edges of the bottle jacks bear the peripheries of the bottle-plugging holes; each sample bottle and each reference bottle are provided with a bottle cork which is plugged with a temperature detection sensing probe in a penetrating manner; and a data transmission cable of each temperature detection sensing probe is connected with an input terminal of the central processing unit. The invention can rapidly detect the quantity and types of microbes in a detected sample by metabolic thermal effect, has the advantages of high precision and efficiency, can be widely used in detection of foods, human body tissues and blood microbes, also can be used in microbe detection of articles, such as skin care product, tableware and the like.
Description
Technical field
The present invention relates to a kind of microorganism detection equipment and detection method.
Background technology
In today of 21 century, along with rapid economic development of China, wide variation have taken place in living standards of the people, and the kind and the form of diet are more rich and varied.But the problem of food quality and security also becomes increasingly conspicuous.Utilize more effective and analysis means fast, set up reliable food safety monitoring system, ensure the healthy of the people, become the work of governments at all levels and relevant department's task of top priority.Food Fast Detection Technique of new generation and product that research and development has independent intellectual property right also become China's researcher obligation one can't decline.
The edible disease that is caused by the food of microbial contamination is called food origin disease.Along with changes of environment and antibiotic abuse, though people's inhabitation and sanitary condition are constantly improved, in continuous decline, food origin disease is in rising trend always to the resistibility of germ.Therefore, also just apparent more important to the detection of pathogenic bacterium in the food.Conventional detection relies on the training objective method of microorganism to determine whether food is polluted mostly, incubation time at least 48 hours, and because colony counts is calculated in range estimation, and it is consuming time not only to require great effort, error is also bigger.Simultaneously, also special laboratory must be arranged, operate under aseptic condition by the professional and technical personnel and just can finish.This is for ten hundreds of middle-size and small-size foodstuff production and processing enterprise, and difficulty is very big.The sample that extracts can only be delivered to special detection mechanism, wait for a couple of days after, just can obtain the result.Time-consuming expensive, monitoring product quality fully in real time.
At present, the research about fast detection technique of food microorganisms both at home and abroad has two main flow directions: the one, and the improvement of traditional viable count method.Groundwork concentrates on the simplification of working specification, improves culture condition, Development of New Generation sample preparation system and experimental installation (substratum, experiment utensil etc.), and the counting process automatization etc.These class methods can make the detection speed of traditional method and efficient be improved, but degree is limited.
Another kind of research direction is to estimate microbial numbers and kind by variation, especially physics or chemical transformation that the measurement microorganism is taken place in growth and metabolic process.According to the difference that detects principle, this class technology can roughly be divided into following several.
1. impedance method (impedence measurement)
Its principle is to show microbial numbers indirectly by measuring the substratum impedance variations that causes because of microorganism growth.The measuring system of having gone on the market such as the Bactometer of France, going through time when it reaches than the low a certain value of substratum initial ion concentration by measuring bacteriogenic ionic concn also claims detection time (DT) to indicate the sample bacterial number.Be to utilize resistance, electric capacity or resulting impedance are measured three parameter systems of ionic concn.Can detected Bacteriophyta hurdle value be 106-107/milliliter.Another listing product is a Malthus microorganism quick analytic instrument.It utilizes bacterium is electrically charged more small molecules with the macromole metabolism in the substratum, thereby the physical principle that causes electricity to lead increase is come bacterial detection quantity.The time that is the living detectable change of living microorganism amount and electric artificial delivery is inversely proportional to.So, pollute seriously more, detection time (DT) is short more.
The principle that the carbonic acid gas that the proposition recently of domestic University Of Tianjin utilizes bacterial metabolism to produce makes the substratum electricity lead variation detects the technology of microbe population, but does not have formal product to come out as yet.
2.ATP bioluminescence technique (bioluminescence, BL)
All living body biologicals all contain ATP, when contacting with ATP, the luciferase by the preparation of biologies such as fluorescence worm just produces luminescence phenomenon, detect luminous intensity and just can obtain living microorganism quantity indirectly. utilize the detecting instrument of this principle that kind more than ten is arranged, as the AF-100 of Japan, the biological optical measuring instrument of E.I.Du Pont Company.
3. microcalorimetry (microcalorimetry)
It is to estimate microbe population indirectly by measuring the metabolic heat that produces in bacterial growth and the metabolic process, and this method needs pre-rendered heating curve figure and special highly sensitive microtemperature transmitter.
4. dissolved oxygen Shu fine linen is tanned
Japan DAIKIN company develops a kind of electric current that causes because of dissolved oxygen in the microbial consumption substratum by detection and changes the instrument that comes the contained microbe population of indirect calculation sample.
5. radiation method (radio metric)
Be that the radio-labeling of trace is introduced in glucose or other carbohydrate molecule, sugar is utilized and emits the carbonic acid gas of mark during bacterial growth, after deriving from culture apparatus, with the radiation measurement instrument detection of U.S. Johnston company, its exit dose and bacterial number are proportional.
Generally speaking, these class methods can increase substantially detection speed, and increase accuracy of detection and alleviate the testing amount, be developing direction from now on.
The principle of institute of the present invention foundation belongs to the microcalorimetry law areas in principle.Because food contamination is serious more, contained living microorganism quantity is just big more, and the increment of unit time intracellular metabolite heat effect is just big more, and this variation characteristic is relevant with the kind of microorganism.The key that adopts this kind technology to carry out microorganism detection is to be subject to environment and artificial interfering factors influence, is difficult to correctly detect the feeble signal that specified microorganisms discharges in specified time, can not address the above problem and still there is test set so far.
Summary of the invention
The purpose of this invention is to provide a kind of microbe rapid detection device and detection method thereof, solve existing microorganism detection method complexity, the measuring accuracy of test set is subject to environment and the influence of artificial interfering factors, and device structure complexity, technical problem that cost is high.
For achieving the above object, the present invention adopts following technical scheme:
This microbe rapid detection device comprises the sample bottle and the reference bottle that detect in box and the box, in sample bottle and the reference bottle detected sample and aseptic object of reference is housed respectively;
Detect box: be an airtight box body that is connected with vacuum extractor with heating unit, it is stacked as one by inner box, insulating sandwich and outer box cover, the end face of described inner box is distributed with inserts the bottle hole, sample bottle and reference bottle insert in the inner box, its bottleneck outer support is being inserted bottle hole periphery, each sample bottle and reference bottle are furnished with bottle stopper, pass bottle stopper and are inserted with the probe temperature sensing probe, and the data cable of each probe temperature sensing probe is connected with the test data input terminal of central processing unit;
The sidewall of inner box and bottom surface are connected with a pair of catheter and a pair of airway respectively, and above-mentioned catheter and airway all are connected with magnetic valve, and liquid-inlet pipe is communicated with the fluid heat storage can of heating unit;
Circulation power pump liquid outlet in liquid discharge pipe and the fluid heat storage can is communicated with;
The extraction pipe of box bottom surface is communicated with the vacuum pump of vacuum extractor;
The eduction pipe of box sidewall passes outer box and is connected with atmosphere;
Heating unit: have in one fluid is housed, the fluid heat storage can of well heater and circulation power pump, described fluid heat storage can has lagging casing, two ends have fluid inlet and fluid outlet respectively up and down, the fluid heat storage can has lagging casing, two ends have fluid inlet and fluid outlet respectively up and down, the circulation power pump is installed on the bottom of fluid heat storage can, fluid is built-in with the fluid temperature (F.T.) probe, the data line of fluid temperature (F.T.) probe is connected with fluid temperature (F.T.) data input pin of central processing unit, and the starting switch of well heater and circulation power pump is connected with the control terminal of central processing unit through control line respectively;
Vacuum extractor: the vacuum pump intake is communicated with the extraction pipe of box bottom surface, and venting port is communicated with atmosphere, and the motor start switch that drives vacuum pump work is connected with a control signal output terminal of central processing unit through control line;
Central processing unit: comprise storer, arithmetical unit, timer, controller, Data Input Interface, control signal output interface and communication interface;
Storer is used for storage operation program, trace routine, nominal data storehouse and detects data;
Arithmetical unit be used for will input each temperature detection data and the nominal data storehouse compare, analyze, calculate, handle and draw detected result and controlling party to;
Controller is used for to well heater output control information, control heater operation, and the keying of Control Circulation power-driven pump, vacuum pump, liquid outlet electromagnetic valve, liquid inlet electromagnetic valve, bleed magnetic valve and pressure-releasing electromagnetic valve;
Communication interface is connected with host computer by communication line;
Power module: output terminal is connected with central processing unit with the motor of well heater, circulation power pump, driving vacuum pump respectively by supply lead, and input terminus is connected with external source;
Liquid crystal display: be connected with the demonstration output terminal of central processing unit, be used for display operation prompting and each parts running status;
Action button: be connected with data input pin of central processing unit, be used for each parts operation of Artificial Control.
The end face of described inner box insert the bottle hole around and outer box cover around be lined with rubber pad, bottle stopper, probe temperature sensing probe, the insulating sandwich of inner box top and the loam cake of outer box are fixed as one, and form and seal lid, seal lid above be connected with handle.
A described sample bottle and a reference bottle outside surface scribble the radioprotective heat-transfer matcrial.
Described heating unit and vacuum extractor, power module and central processing unit are contained in the shell, and liquid crystal display and action button are embedded on the shell, also leave the socket that connects power supply on the shell.
The fluid heat storage can of described heating unit has lagging casing, and two ends have fluid inlet and fluid outlet respectively up and down.
The method for rapid inspecting animalcule of this microbe rapid detection device, the bottle that detected sample and object of reference are housed is put into same airtight detection box, object of reference is identical with sample material, and the material of the microorganism that obtains after adopting the means that can not change the object of reference physical properties thoroughly to sterilize, make the starting temperature of the two reach consistent before the detection earlier, and in detecting box, keep constant temperature, in detection, avoid the heat of each bottle to scatter and disappear and bottle between the heat transmission, utilize the fluid temperature (F.T.) probe that inserts in each bottle in for some time, to gather the temperature signal that transmits, and utilize computer drawing to go out temperature-time changing curve in each bottle, deduction object of reference temperature-time changing curve part from sample temperature-time changing curve, and with computer in existing microorganism Wen-Du time behavior curve of storing compare, draw contained microbe population of sample and kind.
Concrete detection step is as follows:
Well heater in the step 2, fluid heat storage can begins preheating;
Step 5 per sample with the initial temperature of object of reference, is set the preset temp of fluid heat storage can inner fluid;
Step 7, treat each bottle temperature unanimity after, extract the box inner fluid out;
Step 8 is closed pressure-releasing electromagnetic valve, opens the magnetic valve of bleeding, and opens vacuum pump, makes to detect box maintenance constant vacuum;
Step 9, beginning are gathered in real time the temperature signal that the probe temperature sensing probe of each bottle transmits, and utilize computer drawing to go out temperature-time changing curve in each bottle;
The fluidic temperature is than putting into the sample that detects box and the high 5-10 of initial top temperature ℃ of object of reference in the described step 5, the highlyest is no more than 45 ℃.
Fluid and bottle surface are nonwetting in the described step 6, and fluid adopts mineral oils.
Vacuum ranges can be the 0.01-0.95 normal atmosphere in the described step 8.
The computation process that detects total number of bacterial colony in the described step 10 is:
If the sample bottle starting temperature is Ty0, be Tc0 with reference to the bottle starting temperature, sample bottle t temperature constantly is Tyt, is Tct with reference to bottle t temperature constantly, then begins to t constantly the clean temperature increment of sample bottle from detection:
ΔTyt＝(Tyt-Ty0)-(Tct-Tc0)；
Reasonable in design when test set, parameter is selected appropriate, operation just often, Ty0 and Tc0 ten minutes are approaching, following formula is reduced to:
ΔTyt＝Tyt-Tct；
Δ Tyt that obtains and the nominal data that is stored in the computer are compared, the size of supposing Δ Tyt is arranged under the database similarity condition, between two adjacent data Δ Ty1 of similar sample and the Δ Ty2 (Δ Ty1＜Δ Ty2), their corresponding total number of bacterial colony are respectively J1 and J2, then the total number of bacterial colony of this test sample:
Jy＝J1+(ΔTyt-ΔTy1)(J2-J1)/(ΔTy2-ΔTy1)。
Compared with prior art the present invention has following characteristics and beneficial effect:
The present invention has adopted special detection mode and device, a large amount of environment and artificial interfering factors have been got rid of, correctly detect the feeble signal that specified microorganisms discharges in specified time, by a series of suitable amplifications, filter is made an uproar and conversion regime, finally obtain microbe population (total number of bacterial colony) accurately, and identify its kind.
The present invention has following characteristics:
(1), directly detect metabolic effect, rather than detect physics or chemical transformation that meta-bolites produces predetermined substance.The improvement that the simplification of the raising of accuracy of detection and equipment unit is all brought matter.
(2), specially designed probe and equipment can make the faintest metabotic change also can be detected.
(3), custom-designed object of reference, can make environment and artificial interference factor drop to minimum.
(3), specially designed hard, the metabolic characteristic that software system can the accurate recording specified microorganisms, for the discriminating of microorganism provides may.
(4), real-time data acquisition scheme, full-automatic operation and information processing manner, advanced man-machine interface design.
The present invention compares with traditional detection method and existing external method for quick, has following competitive edge:
(1), fast, shorten to 1 to 2 hour or shorter 48 hour detection time that present technique can be required with traditional way.
(2), high precision, minimum bacterial count door hurdle value that can external all kinds of specialized equipment are required significantly reduces (1/2 or lower).Can be low to moderate 1-2/gram (milliliter) by detected colony number.
(3), high-level efficiency, this product can detect dozens of simultaneously, even up to a hundred samples.
(4), easy to use, whole detection system is portable casing, the non-specialised staff just can operate also and can use at the scene through short-term cultivation.
(5), the automatization principle, adopt advanced electronics and IT technology, make system have higher automatization and information processing manner.
(6), do not have easily consumption part or material, be different from most detecting instruments both at home and abroad, present device does not need the material of any frequent replacing, medicine, test paper or part.
The present invention can adopt the influence of the method removal envrionment temperature of establishing reference substance by microbe population and kind in the metabolic heat effect rapid detection sample.Adopt the fluid circulation means to reach sample bottle starting temperature unanimity, adopt vacuum and antiradiation coating method to reduce to conduct heat between bottle, adopt with the correlated method of National Standard Method heat effect is converted to total number of bacterial colony.Adopt with the correlated method of preset temp time curve and determine microbe species.In the test set, the sample detection box is used to settle sample bottle and with reference to bottle, and makes the starting temperature unanimity before the formal detection, prevent as far as possible in detection that heat scatters and disappears and bottle between the heat transmission.Main control system provides the interior predetermined vacuum level of box in consistent required fluid of starting temperature and the testing process; Receive in real time sample bottle and with reference to the temperature analog signal of bottle,, after filter is made an uproar and amplified, send into computer through digital-to-analog conversion.Computer Analysis and these data of processing, by the contrast of sample and reference substance, find out the true metabolic heat feature of microorganism, compare with the microbial metabolism characteristic that is pre-stored in the Computer Database again, obtain the judgement of microbe population and kind, finally show and print.Have precision height, characteristics that efficient is high, can be widely used in the detection of food, tissue and blood microorganism, also can be applicable to skin care product, the microorganism detection of articles for use such as tableware.
Description of drawings
The present invention will be further described in detail below in conjunction with accompanying drawing.
Fig. 1 is the structural representation of microbe rapid detection device.
Fig. 2 detects sample hose in the box and with reference to the layout viewing of pipe.
Fig. 3 detects sample hose in the box and with reference to the layout viewing of pipe.
Fig. 4 detects sample hose in the box and with reference to the layout viewing of pipe.
Fig. 5 detects the real time temperature-time plot that obtains.
Reference numeral: 1-inner box, the 2-fluid, the 3-rubber pad, 4-inserts the bottle hole, 5-probe temperature sensing probe, the 6-bottle stopper, the 7-liquid inlet electromagnetic valve, the 8-liquid outlet electromagnetic valve, the 9-magnetic valve of bleeding, the 10-pressure-releasing electromagnetic valve, the outer box of 11-, the 12-insulating sandwich, the 13-handle, the 14-liquid discharge pipe, the 15-extraction pipe, the 16-liquid-inlet pipe, the 17-fluid inlet, the 18-fluid outlet, 19-circulation power pump, the 20-well heater, 21-fluid temperature (F.T.) probe, the 22-central processing unit, 23-fluid heat storage can, the 24-vacuum pump, the 25-data cable, the 26-liquid crystal display, the 27-action button, the 28-power module, the 29-socket, the 30-direct current supply line, the 31-control line, the 32-communication interface, the 33-host computer, the 34-communication line, the 35-eduction pipe, the 36-shell, the 37-sample bottle, 38-is with reference to bottle.
Embodiment
Embodiment is referring to shown in Figure 1, and this microbe rapid detection device comprises the sample bottle 37 and the reference bottle 38 that detect in box and the box, in sample bottle and the reference bottle detected sample and aseptic object of reference is housed respectively.
Detect box: be an airtight box body that is connected with vacuum extractor with heating unit, it is stacked as one by inner box 1, insulating sandwich 12 with outer box 11 covers.Insulating sandwich 12 is fixed with one with outer box, and insulating sandwich can further reduce the interference of envrionment temperature, can be used on the conventional lagging material (as porous plastics) that uses in refrigerator or the insulation can and makes.The end face of inner box is distributed with inserts bottle hole 4, insert the bottle hole around and outer box cover be lined with rubber pad 3 all around, after the sample bottle of band convex edge inserts inner box, as being negative pressure in the compartmentalized box for holding assorted fruits and candies, then will keep closely contacting between convex edge and the rubber pad, and the box internal pressure is low more, contacts tight more, the certain negative pressure of easy more maintenance (or vacuum tightness) in the box, a sample bottle and a reference bottle outside surface scribble the radioprotective heat-transfer matcrial.Inner box wall 2 can be made by adiabatic mechanically resistant material (as tetrafluoroethylene), can stand negative pressure in certain box and indeformable, sample bottle or with reference to the bottle by heat insulating ability preferably material (as glass) make, outside surface scribbles the material (as silver or aluminium) that radioprotective is conducted heat, can be by electroplating, spray or be similar to the manufacture craft of the glass liner of a thermos flask, even common silver mirror reaction is made.Sample bottle and reference bottle insert in the inner box, and its bottleneck outer support is being inserted a bottle hole periphery, and each sample bottle and reference bottle are furnished with bottle stopper 6.Bottle stopper 6 is made by adiabatic good elasticity material (as rubber or cork), can tightly contact with bottleneck, prevents that bottle interior heat scatters and disappears.Pass bottle stopper and be inserted with probe temperature sensing probe 5, the data cable 25 of each probe temperature sensing probe is connected with the test data input terminal of central processing unit 22.Bottle stopper 6, probe temperature sensing probe 5, the insulating sandwich of inner box top and the loam cake of outer box are fixed as one, and form the sealing lid, are connected with handle 13 above the sealing lid.
The sidewall of inner box and bottom surface are connected with a pair of catheter and a pair of airway respectively, and above-mentioned catheter and airway all are connected with magnetic valve, and liquid-inlet pipe 16 is communicated with the fluid heat storage can 23 of heating unit.The liquid discharge pipe 14 and liquid-inlet pipe 16 outsides are wrapped with thermal insulation layer.
The extraction pipe of box bottom surface is communicated with the vacuum pump of vacuum extractor.
The eduction pipe of box sidewall passes outer box and is connected with atmosphere.
Heating unit: the fluid heat storage can 23 that fluid 2, well heater 20 and circulation power pump 19 are housed in is arranged, and fluid heat storage can 23 is surrounded by lagging casing, and two ends have fluid inlet 17 and fluid outlet 18 respectively up and down.If adopt liquid (as water or oil) as heat-transfer medium, can add liquid by fluid inlet.If directly adopt air as medium.This mouthful can be closed.To use fluid too of a specified duration to emit by fluid outlet, so that change to new heat-transfer medium.
Vacuum extractor: the suction port of vacuum pump 24 is communicated with the extraction pipe 15 of box bottom surface, and venting port is communicated with atmosphere, and the motor start switch that drives vacuum pump work is connected with a control signal output terminal of central processing unit 22 through control line.
Central processing unit 22: comprise storer, arithmetical unit, timer, controller, Data Input Interface, control signal output interface and communication interface.
Storer is used for storage operation program, trace routine, nominal data storehouse and detects data.
Arithmetical unit be used for will input each temperature detection data and the nominal data storehouse compare, analyze, calculate, handle and draw detected result and controlling party to.
Controller is used for to well heater output control information, control heater operation, and the keying of Control Circulation power-driven pump 19, vacuum pump 24, liquid outlet electromagnetic valve 8, liquid inlet electromagnetic valve 7, bleed magnetic valve 9 and pressure-releasing electromagnetic valve 10.
Power module 28: output terminal is connected with central processing unit 22 with the motor of well heater 20, circulation power pump 19, driving vacuum pump respectively by supply lead, and input terminus is connected with external source.
Liquid crystal display 26: be connected with the demonstration output terminal of central processing unit 22, be used for display operation prompting and each parts running status.
Action button 27: be connected with data input pin of central processing unit 22, be used for each parts operation of Artificial Control.
Described heating unit and vacuum extractor, power module 28 and central processing unit 22 are contained in the shell 36, and liquid crystal display 26 and action button 27 are embedded on the shell, also leave the socket 29 that connects power supply on the shell.The prompting of liquid crystal display 26 display operations and each parts (detecting box, vacuum pump, heat storage can) running status.
Because of the present invention adopts physical method indirect detection microbe species and quantity, thus biological method (National Standard Method) contrast that must and directly detect, could precise quantification.Correlated result will import in the database of computer as data in advance.This process is called demarcation.Have two in the demarcation:
(1) total number of bacterial colony is demarcated.Select same sample, be divided into two parts of equivalent after the even processing.Portion carries out biological process (National Standard Method) and detects (dilution back insulation can was cultivated 48 hours, took advantage of extension rate after the microscopically range estimation), obtains this sample total number of bacterial colony.Such as 300/milliliter.Second half sample enters the detection case of the present invention's design simultaneously, presses the real-time collecting temperature data of predetermined process, in the Preset Time that record (detects air pressure and starting temperature) under certain condition, and the pure increased value of sample temperature (having deducted ambient temperature effect).Such as vacuum tightness (see below and state) in detecting box is 0.1 normal atmosphere, and when detecting starting temperature and being 30 ℃, sample temperature pure increased value after 1.0 hours is 0.15 ℃, has increased by 0.17 ℃ after 1.5 hours, has increased by 0.195 ℃ etc. after 2.0 hours.Deposit these data in computer.When detecting other sample later on, keep same air pressure and starting temperature condition, if sample has increased same temperature value in the same time, then contained total number of bacterial colony is identical, promptly is all 300/milliliter.Consider that total number of bacterial colony can not be carried out from big to small infinite time demarcates, can select classification to carry out, promptly select total number of bacterial colony at 1-10,10-50,50-100,100-150,150-200,200-250,250-300,300-400,400-500 (individual/milliliter) .... each sample in the scope is demarcated.Adopt method of interpolation to go out the total number of bacterial colony value again by computed in software.(seeing below calculation formula).Certainly, classification is thin more, and error can be more little, but workload in advance is also big more.In principle, different types of sample need be set up different nominal data storehouses.
(2) biological species is demarcated.Select the special single microorganism of cultivating to add in the aseptic sample, also need add nutrition when needing.Be divided into equivalent after the homogenizing two minutes.Same by two kinds of methods detections.Write down under certain condition, the temperature increase curve of sample after the deduction ambient temperature effect, analytic curve mathematical feature (rate of rise, flex point, asymptotic line or the like) is recorded in the computer in the lump.When under similarity condition, detecting later on, run into when identical curve occurring, just can conclude and contain this kind microorganism in the sample, its quantity and equating that biological process is measured.Certainly, real sample may contain more than a kind of microorganism, and this needs a large amount of previous works, and the xenogenesis microorganism that has been pre-mixed different ratios is demarcated.Set up thousands of curves, just can obtain better result.
The method for rapid inspecting animalcule of this microbe rapid detection device, it is as follows specifically to detect step:
Well heater in the step 2, fluid heat storage can begins preheating.
Step 5 per sample with the initial temperature of object of reference, is set the preset temp of fluid heat storage can inner fluid.The fluidic temperature is than putting into the sample that detects box and the high 5-10 of initial top temperature ℃ of object of reference, the highlyest is no more than 45 ℃.
Step 7, treat each bottle temperature unanimity after, extract the box inner fluid out.
Step 8 is closed pressure-releasing electromagnetic valve, opens the magnetic valve of bleeding, and opens vacuum pump, makes to detect box maintenance constant vacuum; Vacuum ranges can be the 0.01-0.95 normal atmosphere.In testing process, remain the certain vacuum degree in the inner box, can keep the thermal insulation between bottle and the bottle, also the influence of envrionment temperature is reduced to minimum.
Step 9, beginning are gathered in real time the temperature signal that the probe temperature sensing probe of each bottle transmits, and utilize computer drawing to go out temperature-time changing curve in each bottle.
The computation process of described detection total number of bacterial colony is:
If the sample bottle starting temperature is Ty0, be Tc0 with reference to the bottle starting temperature, sample bottle t temperature constantly is Tyt, is Tct with reference to bottle t temperature constantly, then begins to t constantly the clean temperature increment of sample bottle from detection:
ΔTyt＝(Tyt-Ty0)-(Tct-Tc0)；
Reasonable in design when test set, parameter is selected appropriate, operation just often, Ty0 and Tc0 ten minutes are approaching, following formula is reduced to:
ΔTyt＝Tyt-Tct；
Δ Tyt that obtains and the nominal data that is stored in the computer are compared, the size of supposing Δ Tyt is arranged under the database similarity condition, between two adjacent data Δ Ty1 of similar sample and the Δ Ty2 (Δ Ty1＜Δ Ty2), their corresponding total number of bacterial colony are respectively J1 and J2, then the total number of bacterial colony of this test sample:
Jy＝J1+(ΔTyt-ΔTy1)(J2-J1)/(ΔTy2-ΔTy1)。
When a beginning new round detects, can change the new bottle after the sterilization, use sterilized instrument that sample and object of reference are added in the new bottle, make probe and the corresponding bottleneck of each autoregistration of bottle stopper after the sterilization again, loam cake is relay down and pressurize, each bottle stopper and bottleneck are sealed fully, and allow probe enter in sample or the object of reference.By each magnetic valve of abovementioned steps opening and closing, inject the equal gentleness of fluid again and vacuumize in detecting box then, a beginning new round detects.After loam cake puts in place, should guarantee closely to contact with on every side thermofin.To reduce the influence of envrionment temperature to greatest extent.
Referring to shown in Figure 24, detect sample hose in the box and have two with reference to the arrangement principle of pipe:
(1) the least possible with reference to the quantity of pipe, in order to avoid take too much space.
(2) should be representative as far as possible with reference to the position of pipe, to avoid the interference difference of envrionment temperature to different positions.
Pipe position among Fig. 2 is layout ringwise.From detecting the temperature effect of box surrounding temperature to each pipe the box, each bottle position is equity fully, and 1 place gets final product as the reference bottle with a bottle so only need at an arbitrary position.
Pipe position among Fig. 3 is square layout because surrounding temperature is incomplete same to the influence of each position, so the bottle of getting 1 and 2 positions as with reference to bottle, with the average back of two bottles temperature datas as the data of object of reference enter calculate proper.
For more bottles and more complicated layout, be preferably in the weighing apparatus of making even between the two principle among Fig. 4.Both taken into full account the difference of different positions, the least possible again establishing with reference to bottle.Can select center and peripheral each one bottle, promptly place with reference to bottle 3 and 4 positions.
The detection example of microbe rapid detection device is as follows:
Adopt present device, obtain following detected result according to aforementioned detection step: sample is certain brand high-temperature sterilization milk (also having 1 month quality guaranteed period to expire).Object of reference is same bag milk, but ultraviolet disinfection 25 minutes.Sample and object of reference are 45 milliliters.Ambient temperature range 25-28 ℃.The fluid preset temp is 23.05 ℃.Vacuum tightness is 0.2 normal atmosphere in the box.The sample bottle starting temperature is 23.04 ℃, is 23.045 ℃ with reference to the bottle starting temperature.Real time temperature-time curve that detection obtains as shown in Figure 5.Ordinate zou representation temperature among Fig. 5 (℃), X-coordinate is represented the time (point represented for 4 seconds).Rising is the sample bottle temperature curve faster, and slower is with reference to the bottle temperature curve.As shown in Figure 5, higher because of room temperature, envrionment temperature is still influential to detecting box.After adopting this influence of aforementioned formula (1) and (2) deduction, sample bottle is 0.15 in the clean temperature increment of formal detection after 1 hour.
Look into the nominal data storehouse of this brand milk under the similarity condition, finding to rise in value with 0.15 ℃ of two nearest adjacent temperature is 0.12 ℃ and 0.20 ℃, corresponding total number of bacterial colony is respectively 155 and 370 (individual/milliliter), and by aforementioned calculation formula (3), the total number of bacterial colony that obtains this sample is:
J＝155+(0.15-0.12)(370-155)/(0.20-0.12)
=236 (individual/milliliter).
Same sample detects with National Standard Method, and the total number of bacterial colony that obtains is 250/milliliter, and is very approaching with The above results.Confirmed the reasonableness of the technology of the present invention and equipment design.Same comparison and detection instrument carries out repeatedly.Can repeat to obtain similar results.
Claims (10)
1. a microbe rapid detection device comprises the sample bottle 37 and the reference bottle 38 that detect in box and the box, in sample bottle and the reference bottle detected sample and aseptic object of reference is housed respectively, it is characterized in that:
Detect box: be an airtight box body that is connected with vacuum extractor with heating unit, it is stacked as one by inner box 1, insulating sandwich 12 with outer box 11 covers, the end face of described inner box is distributed with inserts bottle hole 4, sample bottle and reference bottle insert in the inner box, its bottleneck outer support is being inserted bottle hole periphery, each sample bottle and reference bottle are furnished with bottle stopper (6), pass bottle stopper and be inserted with probe temperature sensing probe (5), the data cable of each probe temperature sensing probe (25) is connected with the test data input terminal of central processing unit (22);
The sidewall of inner box and bottom surface are connected with a pair of catheter and a pair of airway respectively, and above-mentioned catheter and airway all are connected with magnetic valve, and liquid-inlet pipe (16) is communicated with the fluid heat storage can (23) of heating unit;
Circulation power pump (19) liquid outlet in liquid discharge pipe (14) and the fluid heat storage can is communicated with;
The airway of box bottom surface (15) is communicated with the vacuum pump (24) of vacuum extractor;
The airway of box sidewall (35) passes outer box (11) and is connected with atmosphere;
Heating unit: the fluid heat storage can (23) that fluid (2), well heater (20) and circulation power pump (19) are housed in is arranged, described circulation power pump (19) is installed on the bottom of fluid heat storage can, fluid (2) is built-in with fluid temperature (F.T.) probe (21), the data line of fluid temperature (F.T.) probe (21) is connected with the fluid temperature (F.T.) data input pin of central processing unit (22), and the starting switch of well heater (20) and circulation power pump (19) is connected through the control terminal of control line (31) with central processing unit (22) respectively;
Vacuum extractor: the suction port of vacuum pump (24) be communicated with the airway (15) of box bottom surface, venting port is communicated with atmosphere, the motor start switch that drives vacuum pump work is connected with a control signal output terminal of central processing unit (22) through control line;
Central processing unit (22): comprise storer, arithmetical unit, timer, controller, Data Input Interface, control signal output interface and communication interface;
Storer is used for storage operation program, trace routine, nominal data storehouse and detects data;
Arithmetical unit be used for will input each temperature detection data and the nominal data storehouse compare, analyze, calculate, handle and draw detected result and controlling party to;
Controller is used for to well heater output control information, control heater operation, and the keying of Control Circulation power-driven pump (19), vacuum pump (24), liquid outlet electromagnetic valve 7, liquid inlet electromagnetic valve 8, bleed magnetic valve 9 and pressure-releasing electromagnetic valve 10;
Communication interface (32) is connected with host computer (33) by communication line (34);
Power module (28): output terminal is connected with central processing unit (22) with the motor of well heater (20), circulation power pump (19), driving vacuum pump respectively by supply lead, and input terminus is connected with external source;
Liquid crystal display (26): be connected with the demonstration output terminal of central processing unit (22), be used for display operation prompting and each parts running status;
Action button (27): be connected with data input pin of central processing unit (22), be used for each parts operation of Artificial Control.
2. microbe rapid detection device according to claim 1, it is characterized in that: the end face of described inner box insert bottle hole (4) around and outer box cover be lined with rubber pad (3) all around, bottle stopper (6), probe temperature sensing probe (5), the insulating sandwich of inner box top and the loam cake of outer box are fixed as one, form the sealing lid, be connected with handle (13) above the sealing lid.
3. microbe rapid detection device according to claim 1 is characterized in that: a described sample bottle and a reference bottle outside surface scribble the radioprotective heat-transfer matcrial.
4. microbe rapid detection device according to claim 1, it is characterized in that: described heating unit and vacuum extractor, power module (28) and central processing unit (22) are contained in the shell (36), liquid crystal display (26) and action button (27) are embedded on the shell, also leave the socket (29) that connects power supply on the shell.
5. an application rights requires the method for rapid inspecting animalcule of any microbe rapid detection device of 1-4, it is characterized in that: the bottle that detected sample and object of reference will be housed is put into same airtight detection box, object of reference is identical with sample material, and the material of the microorganism that obtains after adopting the means that can not change the object of reference physical properties thoroughly to sterilize, make the starting temperature of the two reach consistent before the detection earlier, and in detecting box, keep constant temperature, in detection, avoid the heat of each bottle to scatter and disappear and bottle between the heat transmission, utilize the fluid temperature (F.T.) probe that inserts in each bottle in for some time, to gather the temperature signal that transmits, and utilize computer drawing to go out temperature-time changing curve in each bottle, deduction object of reference temperature-time changing curve part from sample temperature-time changing curve, and with computer in existing microorganism temperature-time behavior curve of storing compare, draw contained microbe population of sample and kind.
6. method for rapid inspecting animalcule according to claim 5, it is as follows to it is characterized in that specifically detecting step:
Step 1 is opened the switch of microbe rapid detection device, selects detection time according to the sample bacteria containing amount of pre-estimating;
Well heater in the step 2, fluid heat storage can begins preheating;
Step 3 pulls up the sealing lid that detects box, inserts sample bottle and with reference to bottle, is the material that does not contain microorganism identical with sample material, that adopt physical method thoroughly to obtain after the sterilization with reference to the object of reference in the bottle;
Step 4, probe temperature sensing probe and bottle stopper carried out sterilising treatment after, aim at corresponding bottleneck, the sealing cover body weight is newly put down and pressure seal;
Step 5 per sample with the initial temperature of object of reference, is set the preset temp of fluid heat storage can inner fluid;
Step 6, treat that fluid reaches preset temp after, open box sidewall liquid-inlet pipe and liquid inlet electromagnetic valve, start the circulation power pump, the fluid in the fluid heat storage can is injected the inner box that detects box, circulation samming;
Step 7, treat each bottle temperature unanimity after, extract the box inner fluid out;
Step 8 is closed pressure-releasing electromagnetic valve, opens the magnetic valve of bleeding, and opens vacuum pump, makes to detect box maintenance constant vacuum;
Step 9, beginning are gathered in real time the temperature signal that the probe temperature sensing probe of each bottle transmits, and utilize computer drawing to go out temperature-time changing curve in each bottle;
Step 10, utilize computer calculates detection total number of bacterial colony and judge microbe species, deduction object of reference temperature variation curve part from the sample temperature change curve, and with computer in existing microorganism temperature-time behavior curve compare, directly demonstrate contained microbe population of sample and kind;
Step 11 after detection is finished, is opened pressure-releasing electromagnetic valve, allows detect in the box return to normal pressure, opens loam cake, takes out sample bottle and reference bottle.
7. method for rapid inspecting animalcule according to claim 6 is characterized in that: the fluidic temperature the highlyest is no more than 45 ℃ than putting into the sample that detects box and the high 5-10 of initial top temperature ℃ of object of reference in the described step 5.
8. microbe rapid detection device according to claim 6 is characterized in that: fluid and bottle surface are nonwetting in the described step 6, and fluid adopts mineral oils.
9. method for rapid inspecting animalcule according to claim 6 is characterized in that: vacuum ranges is the 0.01-0.95 normal atmosphere in the described step 8.
10. method for rapid inspecting animalcule according to claim 6 is characterized in that: the computation process that detects total number of bacterial colony in the described step 10 is:
If the sample bottle starting temperature is Ty0, be Tc0 with reference to the bottle starting temperature, sample bottle t temperature constantly is Tyt, is Tct with reference to bottle t temperature constantly, then begins to t constantly the clean temperature increment of sample bottle from detection:
ΔTyt＝(Tyt-Ty0)-(Tct-Tc0)；
Reasonable in design when test set, parameter is selected appropriate, operation just often, Ty0 and Tc0 ten minutes are approaching, following formula is reduced to:
ΔTyt＝Tyt-Tct；
Δ Tyt that obtains and the nominal data that is stored in the computer are compared, the size of supposing Δ Tyt is arranged under the database similarity condition, between two adjacent data Δ Ty1 of similar sample and the Δ Ty2 (Δ Ty1＜Δ Ty2), their corresponding total number of bacterial colony are respectively J1 and J2, then the total number of bacterial colony of this test sample:
Jy＝J1+(ΔTyt-ΔTy1)(J2-J1)/(ΔTy2-ΔTy1)。
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| CN2009103095765A CN101701184B (en) | 2009-11-12 | 2009-11-12 | Microbe rapid detection device and detection method thereof |
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| CN101701184B CN101701184B (en) | 2012-07-11 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103361266A (en) * | 2013-07-12 | 2013-10-23 | 昆山爱达斯工业设计有限公司 | Rapid multi-parameter microorganism detector |
| CN105278375A (en) * | 2015-10-16 | 2016-01-27 | 珠海迪尔生物工程有限公司 | Intelligent automatic microbial detection device and method |
| CN108315231A (en) * | 2018-01-26 | 2018-07-24 | 楼德耀 | A kind of new bio detection device |
| CN109097433A (en) * | 2018-09-25 | 2018-12-28 | 江西中医药大学 | Content of microorganisms detection method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20050250138A1 (en) * | 2004-05-04 | 2005-11-10 | Millipore Corporation | Process for the enumeration and identification of microorganisms |
| CA2607086C (en) * | 2005-05-05 | 2015-11-17 | Ravi Kanipayor | System for rapid analysis of microbiological materials in liquid samples |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103361266A (en) * | 2013-07-12 | 2013-10-23 | 昆山爱达斯工业设计有限公司 | Rapid multi-parameter microorganism detector |
| CN105278375A (en) * | 2015-10-16 | 2016-01-27 | 珠海迪尔生物工程有限公司 | Intelligent automatic microbial detection device and method |
| CN108315231A (en) * | 2018-01-26 | 2018-07-24 | 楼德耀 | A kind of new bio detection device |
| CN109097433A (en) * | 2018-09-25 | 2018-12-28 | 江西中医药大学 | Content of microorganisms detection method and application |
| CN113907820A (en) * | 2021-10-08 | 2022-01-11 | 巢湖市宾雄医疗器械有限公司 | Intelligent suture needle sleeve capable of self-detecting bacteria content and detection method thereof |
| CN113907820B (en) * | 2021-10-08 | 2023-05-30 | 巢湖市宾雄医疗器械有限公司 | Suture needle sleeve for intelligently and automatically detecting bacterial content and detection method thereof |
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