CN109082456A - A kind of automated food microorganism detection pre-treating method and device - Google Patents
A kind of automated food microorganism detection pre-treating method and device Download PDFInfo
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Abstract
The present invention relates to technical field of microbial detection, a kind of automated food microorganism detection pre-treating method and device are disclosed.Cultivation platform, culture cup and the enrichment facility of automation, thermostat and safety guarantee coercive procedure that pre-treating method of the invention passes through automation rotation, bacterium and complete machine safety guarantee is taken to be introduced into food microorganisms pre-treatment washing, farthest avoid quality and the quantity loss by mechanical crushing method bring bacterium, the accuracy for improving detection, the risk for reducing infected person, polluting environment.
Description
Technical field
The present invention relates to Microbiological detection of foods fields, and in particular to a kind of to be extracted the bacterium on sample by water-washing method
The automated food microorganism detection pre-treating method to get off and its corresponding processing unit.
Background technique
Pathogenic bacteria are based on molecular level and detect rapid proliferation in food in recent years.Its detecting step mainly include pre-treatment,
Three nucleic acid extraction, nucleic acid amplification and detection steps.In order to guarantee to extract all pathogenic microorganisms in sample, preceding
In processing, it is common to use sample is mechanically pulverized, such as grinding is squeezed, hit, shearing, then extracts to obtain by modes such as filterings
Pathogenic microorganisms in sample, and then detect the type and quantity of microorganism.The sample pre-treatments mode of this mechanical crushing by
The liquid intracellular generated after mechanical crushing has bactericidal effect, causes the quantity for detecting microorganism practical containing micro organism quantity
It reduces, haves the defects that amount detection is low, to keep testing result inaccurate.
Summary of the invention
The present invention provides a kind of automated food microorganism detection pre-treating method and device, improves the accuracy rate of detection.
Technical scheme is as follows:
A kind of automated food microorganism detection pre-treating method, comprising the following steps:
(1) sample is put into the culture cup for being preinstalled with culture medium, sterile water is added, is placed in the corresponding fixation of cultivation platform
In device；
(2) after culture cup rotation mixes sample and culture medium, cultivation platform and culture cup carry out planetary motion, and sample exists
It is moved in culture cup, extracts the microorganism in sample；
(3) sample in culture cup is held up with strainer, enrichment facility is drawn the culture solution in culture cup and carried out to sample
It rinses, enrichment facility is passed through the culture solution in filtrated air concentration culture cup after strainer decline；Repeat this step；
(4) it shifts and collects concentrate.
Preferably, the culture cup after being put into sample is independent sealed.
Preferably, the method carries out in the confined space for being provided with exhaust gas discharge conduit.
Further, the exhaust gas discharge conduit is connected in culture concentration process with each culture cup.
Further, the culture cup surrounding is provided with thermostat.
Preferably, the temperature of step (2) the thermostat setting is 37 DEG C；Step (3) the thermostat setting
Temperature is 45 DEG C.
Preferably, the culture cup inner wall is provided with protrusion.
Preferably, after concentrate transfer, culture cup is destroyed as medical disposable material.
It preferably, further include cleaning gas and/or pressure sterilizing program before and after detection pre-treatment.
The present invention also provides a kind of automated food microorganism detection pretreating device, including closed main body, Yi Jishe
Be placed in the closed master it is intracorporal can closed independent culture cup, cultivation platform, enrichment facility, thermostat and safety guarantee it is strong
Device processed；The cultivation platform can be equipped with the fixed dress of culture cup relative to the closed horizontal rotation of main body, cultivation platform
It sets, the culture cup is with the fixed horizontal rotation of device；Strainer is provided in culture cup, for lifting sample；The enrichment facility
Imbibition flushing is carried out to sample when strainer rises, enrichment facility is passed through filtrated air and is concentrated after strainer decline；The perseverance
Warm device provides culture and the thickening temperature of needs for the culture cup；The safety guarantee forcing device include ultraviolet radiator and
Sterile air-breather is carried out forcing sterilizing before and after booting by process control and is passed through filtrated air operation.
Compared with prior art, the beneficial effects of the present invention are:
Automation washing is taken bacterium to introduce food microorganisms by automated food microorganism detection pre-treating method of the invention
In pre-treatment, quality and the quantity loss by mechanical crushing method bring bacterium are farthest avoided, the micro- life of food is improved
The accuracy of analyte detection.The present invention is also concentrated while microorganism is washed in sample to containing bacteria culture fluid, into
One step improves the accuracy and susceptibility of detection pattern detection microorganism.
As a kind of preferred embodiment, the present invention is using the individually culture cup containing culture medium as unit of each sample
Encapsulation, items selection is flexible, can place Arbitrary Term purpose culture cup in one experiment, can be same project in culture cup,
Or the culture cup of disparity items, bacterium culture is realized while being proposed, the flexibility of entry detection is substantially increased.
The present invention provides the device for being suitable for above-mentioned detection pre-treating method, cultivation platform and culture cup are mentioned as washing
The storage space of bacterium method, and ensure that each sample has independent processing space, can closed independent culture cup by being embedded with not
With the culture cup of culture medium, solve the problems, such as with batch experiment disparity items.Enrichment facility plays be passed through nothing during processing
Bacterium air is cultivated, and is concentrated using filtrated air, carries out washing bacterium process using culture medium.Thermostat temperature needed for solving to cultivate
Temperature condition needed for further solving concentration under the premise of degree.Safety guarantee forcing device, which fundamentally prevents operation, to be caused
Sterilizing the problems such as being not thorough, give the guarantee of experiment and personnel fundamentally.
The present invention can be realized automation washing using the device and take bacterium, and flexible operation is easy, improves the accuracy of detection.
Washing take bacterium and complete machine safety guarantee to be introduced into food microorganisms pre-treatment, farthest avoid bacterium quality and quantity by
The risk for mechanical crushing method bring loss and reducing infected person, polluting environment.Apparatus of the present invention can pass through vehicular electricity storage
Pond powered operation, have the characteristics that it is portable, achieve the purpose that at any time sample, detect at any time.
Detailed description of the invention
Fig. 1 is automated food microorganism detection pretreating device schematic diagram of the present invention；
Fig. 2 is the partial schematic diagram of automated food microorganism detection pretreating device of the present invention；
Fig. 3 is the culture cup schematic diagram of automated food microorganism detection pretreating device of the present invention；
Wherein, 1, closed main body, 2, culture cup, 3, cultivation platform, 4, enrichment facility, 5, thermostat, 6 safety guarantee are strong
Device processed, 7, fixed device, 8, strainer, 9, exhaust gas discharge conduit, 10, concentrate transfering channel, 11, rinse concentration needle, 12, prominent
It rises；
Fig. 4 is the pre-treating method schematic flow chart that microorganism detection is carried out using apparatus of the present invention.
Specific embodiment
Below by specific embodiment combination attached drawing, invention is further described in detail.
The present invention provides a kind of automated food microorganism detection pre-treating methods, handle before Microbiological detection of foods
In, bacterium, culture, concentration and further preferred complete machine security protection will be proposed and be fused together, it is micro- to realize automated food
The pre-treatment of biological detection improves the accuracy of detection.
Automated food microorganism detection pre-treating method of the present invention, comprising the following steps:
(1) sample is put into the culture cup for being preinstalled with culture medium, appropriate amounts of sterilized water is added, it is corresponding to be placed in cultivation platform
In fixed device；
(2) after culture cup rotation mixes sample and culture medium, cultivation platform and culture cup carry out planetary motion, and sample exists
It is moved in culture cup, extracts the microorganism in sample；
(3) sample in culture cup is held up with strainer, enrichment facility is drawn the culture solution in culture cup and carried out to sample
It rinses, enrichment facility is passed through the culture solution in filtrated air concentration culture cup after strainer decline；Repeat this step；
(4) it shifts and collects concentrate.
Pre-buried culture medium in culture cup of the present invention is put into sample injection appropriate amounts of sterilized water before use.Culture cup
Pilot scale test sample and the difference of target pathogenic bacteria, can be embedded with different culture mediums.Culture cup inner wall has protrusion, facilitates
Turbine is formed convenient for extracting the bacterium contained in sample.
After culture cup of the invention requires sample to be put into, preferably carry out proposing bacterium, culture, concentration under freestanding environment, therefore,
Culture cup after being put into sample is answered independent sealed.
As a kind of specific embodiment, culture cup operating process is:
1) sample is put into culture cup, and culture cup carries out low speed rotation and mixed；
2) after sample blending, cultivation platform and culture cup carry out planetary motion, and sample is not transported at the uniform velocity in culture cup
It is dynamic, to have the function that extract bacterium；
3) culture cup rotation again, preferably rotation time are 15~30min, more preferably 20min.After autobiography stops, culture
Cup in strainer sample is held up, enrichment facility draw culture solution to sample carry out high pressure washing, preferably washing time be 1~
3min；
4) enrichment facility is passed through filtrated air and is concentrated after strainer declines, and concentration time is preferably 2~10min.
The method of the invention carries out in confined space, the confined space contain independent exhaust gas discharge conduit and
Independent and recycling concentrate transfering channel.Wherein, concentrate transfering channel can also be used to be passed through filtrated air, exhaust gas row
It puts channel to be connected in culture concentration process with each culture cup, extra exhaust gas is discharged external.
Thermostat is additionally provided in confined space of the present invention.The thermostat is set to the four of the culture cup
Week, the temperature needed for providing culture and concentration.Preferably, the temperature of the thermostat setting cultivation stage is 37 DEG C,
The temperature that enriching stage is arranged is 45 DEG C.
The enrichment facility of the invention enrichment facility when strainer rises carries out imbibition flushing, and enrichment facility is logical after strainer decline
Enter filtrated air to be concentrated.
After being concentrated into the degree of requirement, the detection that concentrate is used for microorganism is shifted and collected.Pre-treatment terminates, culture cup
After the completion of interior concentrated liquid transfer, culture cup is destroyed as medical disposable material.
As a preferred embodiment, the present invention further includes cleaning gas and/or strong before and after the processing before detection
Sterilizing program processed.The present invention is preferably provided with safety guarantee forcing device, including discrepancy cleaning gas, complete machine are airtight, program is protected
Barrier and sample separate space.
As a preferred embodiment, described program ensures that operating process is:
1) ultraviolet sterilization after instrument is powered, preferably sterilization time are 5~15min, further preferably 10min；It is passed through nothing
Bacterium air is taken a breath, and confined space hatch door can just be opened after movement；
2) sample is put into rear culture cup and carries out independent sealed, ensures the separate space of sample；
3) it cultivates each sample in concentration process and corresponds to independent exhaust gas discharge conduit, handled；
4) culture cup transfer is completed after same day experiment, and before shutdown, confined space hatch door is locked, enforces ultraviolet go out
Bacterium, preferably sterilization time are 20~40min, are further 30min；Lead to 20~40min of filtrated air, is further
30min, air intake vent is closed out.
The present invention also provides a kind of devices suitable for above-mentioned automated food microorganism detection pre-treating method, including
Closed main body, and be set to the closed master it is intracorporal can closed independent culture cup, cultivation platform, enrichment facility, constant temperature dress
It sets and safety guarantee forcing device；The cultivation platform can be relative to the closed horizontal rotation of main body, on cultivation platform
Equipped with the fixed device of culture cup, the culture cup is with the fixed horizontal rotation of device；Strainer is provided in culture cup, for lifting sample
Product；The enrichment facility carries out imbibition flushing to sample when strainer rises, and enrichment facility is passed through filtrated air after strainer decline
It is concentrated；The thermostat provides culture and the thickening temperature of needs for the culture cup；The safety guarantee forces dress
It sets including ultraviolet radiator and sterile air-breather, carry out forcing sterilizing before and after booting by process control and is passed through filtrated air
Operation.
Cultivation platform and culture cup mention the storage space of bacterium method as washing and ensure that each sample has independent processing
Space, can closed independent culture cup by being embedded with the culture cup of different culture medium, solve asking with batch experiment disparity items
Topic.Enrichment facility plays during processing to be passed through filtrated air and cultivates, and is concentrated using filtrated air, is carried out using culture medium
Wash bacterium process.Temperature condition needed for thermostat further solves concentration under the premise of solving and cultivating required temperature.Peace
All risk insurance barrier forcing device fundamentally prevents the problems such as sterilizing caused by operation is not thorough, and gives the guarantor of experiment and personnel fundamentally
Barrier.
Washing is taken bacterium and complete machine safety guarantee to be introduced into food microorganisms pre-treatment by the present invention, is farthest avoided
By quality and the quantity loss of mechanical crushing method bring bacterium and to the risk of personnel, environment.Meanwhile the device is using individually containing
The culture cup of culture medium is encapsulated as unit of each sample, and items selection is flexible, can place Arbitrary Term purpose in one experiment
Culture cup, and bacterium culture is proposed simultaneously, the flexibility of entry detection is substantially increased, extraction, culture and concentration are greatly reduced
Time-consuming, 3 hours can be completed.Automated food microorganism detection pre-treating method of the invention and device have portable spy
Point can be achieved the purpose that sample at any time, be detected at any time by Vehicular accumulator cell powered operation.
In the following, the present invention is by taking food microorganisms mention bacterium as an example to automated food microorganism detection pre-treating method and device
It is illustrated.
It should be understood that these examples are only for illustrating the present invention and are not intended to limit the scope of the present invention.In addition, it should also be understood that,
After having read the content of the invention recorded, those skilled in the art can make various modifications or changes to the present invention, these
Equivalent form is also fallen within the scope of the appended claims of the present application.
Embodiment 1
A kind of automated food microorganism detection pretreating device, including closed main body 1, and it is set to the closed master
In body 1 can closed independent culture cup 2, cultivation platform 3, enrichment facility 4, thermostat 5 and safety guarantee forcing device 6.
The cultivation platform 3 can be consolidated relative to the horizontal rotation of the closed main body 1, several culture cups 2 that are surrounded by of cultivation platform 3
Determine device 7, for culture cup 2 to be fixed on cultivation platform 3, planetary motion can be made with cultivation platform 3.Culture cup 2 is solid
Due in the fixation device 7 on cultivation platform 3, and it can be realized the horizontal rotation of culture cup 2.2 inner wall of culture cup has protrusion 12,
It is embedded with culture solution in culture cup 2, after sample is put into culture cup 2, the closing of culture cup 2 forms separate space.It is set in culture cup 2
It is equipped with strainer 8, for lifting the sample in culture cup 2；It is also communicated with exhaust gas discharge conduit 9 in culture cup and concentrate transfer is logical
Road 10.Enrichment facility 4 is set to inside closed main body 1, is connected with concentrate transfering channel 10.The flushing of enrichment facility 4 is dense
Shortening 11 is placed in inside culture cup 2, and the culture solution drawn in culture cup 2 when strainer 8 rises carries out high pressure washing, filter to sample
Enrichment facility 4 is passed through filtrated air into culture cup 2 by concentrate transfering channel 10 and is concentrated after net 8 declines；Constant temperature dress
52 peripheries of culture cup being set to inside closed main body 1 are set, provide culture and the thickening temperature of needs for the culture cup 2；It is close
It closes inside main body 1 and is additionally provided with safety guarantee forcing device 6, including ultraviolet radiator and sterile air-breather, pass through process control
It carries out forcing sterilizing before and after booting and is passed through filtrated air operation.
Embodiment 2
25g oatmeal is put into the culture cup for being preinstalled with agar medium, 225ml sterile water is added to graduation mark, puts
Enter in the corresponding fixed device of cultivation platform, closes hatch door.Rotor gas circuit automatic butt, filtrated air are passed through, thermostat control
For temperature to 37 DEG C, low speed rotation under culture cup 100rpm mixes sample culturing base.Cultivation platform carries out not at the uniform velocity in culture cup
Planetary motion, minimum speed 100rpm, maximum speed 220rpm.After enrichment facility is passed through filtrated air 10min, by thermostat
To 45 DEG C of 10min, the strainer in culture cup rises temperature control, and it is high that the flushing concentration needle of enrichment facility draws the culture medium in culture cup
Pressure rinses sample 1min, and enrichment facility is passed through filtrated air after strainer decline, and strainer rises after 8min, and ventilation becomes culture medium punching
It washes, pre-treatment is completed after cyclic 3h.Concentrate is shifted, culture cup is destroyed as medical disposable material.Opening program ensures behaviour
Make.
Embodiment 3
25g pork is put into the culture cup for being preinstalled with agar medium, 225ml sterile water is added to graduation mark, is put into
In the corresponding fixed device of cultivation platform, hatch door is closed.Rotor gas circuit automatic butt, filtrated air are passed through, thermostat temperature control
To 37 DEG C, low speed rotation under culture cup 100rpm mixes sample culturing base.Cultivation platform carries out row not at the uniform velocity in culture cup
Luck is dynamic, minimum speed 100rpm, maximum speed 220rpm.Then culture cup carries out the rotation of 20min again.Enrichment facility is passed through
After filtrated air 10min, by thermostat temperature control to 45 DEG C of 10min.Strainer in culture cup rises, and the flushing of enrichment facility is dense
The culture medium high pressure washing sample 2min in culture cup is drawn in shortening, and enrichment facility is passed through filtrated air, 5min after strainer decline
Strainer rises afterwards, and ventilation becomes culture medium flushing, and cyclic 3.5h completes pre-treatment.Concentrate is shifted, culture cup is as one
Secondary property consumptive material is destroyed.Opening program ensures operation.
Embodiment 4
It examines in meat products containing staphylococcus aureus in the standard for washing extraction method and two kinds of extracting modes of mechanical extraction
True property
1, instrument
Rotary cutter type homogenizer
Extraction apparatus is washed shown in the embodiment of the present invention 1
2, bacterial strain
Staphylococcus aureus (Staphylococcus aureus)
3, sample message
Roasted beef, cold cuts flexible.
4, method
Bacterium solution preparation: by staphylococcus aureus secondary culture, 3 generation nutrient agar base inclined-plane fresh cultured objects is taken, 5ml is used
Sterile saline washes down lawn, and the bacteria suspension of experimental concentration is prepared into after being sufficiently mixed.It will be tested with sterile saline
Bacterium solution is diluted to 102~105The bacteria suspension of cfu/ml difference bacteria containing amount measures bacterial content with conventional viable counts method.
The preparation of artificial contamination's sample:
Using from the sample 200g bought on the market, it is soaked in prepared by 500ml 102cfu/ml、103cfu/ml、
104cfu/ml、105In cfu/ml staphylococcus aureus liquid physiological saline, 20min is impregnated, drying is taken out after immersion.
By sample segment be 6 parts, wherein 1,3,5 be group 2,2,4,6 be organize 1.
Group 1: by 25g, treated that sample is put into 225ml physiological saline, and using mechanical crushing method, pulverizer crushes 1min,
The even liquid of sample is made.
Group 2: by 25g, treated that sample is put into 225ml physiological saline, uses present invention washing extraction method, and washing is rinsed
The even liquid of sample is made in 15min.
Blank control group: sample is replaced with 25ml physiological saline respectively, and presses above-mentioned 2 method validations.
(plate count) national standard is examined to test by GB 4789.10-2016 golden yellow color staphylococcus, every kind
Test, each dilution rank prepare 3 plates respectively.Total plate count and each plating amount of mold count are 1ml, clump count
Take the average value of 3 plates.3 plating amounts of each dilution grade of staphylococcus aureus are respectively 0.3,0.3,0.4ml,
Clump count by 3 plate bacterium colonies and count.And bacterium colony numerical value is reported according to the report rule of every kind of bacterium.It the results are shown in Table 1.
Table 1 washes staphylococcus aureus result in extraction method and mechanical extraction detection cooked beef
Food microorganisms pre-treatment automated detection method of the present invention, before taking bacterium to introduce food microorganisms automation washing
In processing, quality and the quantity loss by mechanical crushing method bring bacterium are farthest avoided, food microorganisms are improved
The accuracy of detection.It can be seen from Table 1 that the quantity of washing extraction method measurement staphylococcus aureus more connects through the invention
It is bordering on the actual value of microorganism in sample, reduces the influence in pretreatment process to data result, improves accuracy rate.
Use above specific case is illustrated the present invention, is merely used to help understand the present invention, not to limit
The system present invention.For those skilled in the art, according to the thought of the present invention, can also make several simple
It deduces, deform or replaces.
Claims (10)
1. a kind of automated food microorganism detection pre-treating method, which comprises the following steps:
(1) sample is put into the culture cup for being preinstalled with culture medium, sterile water is added, is placed in the corresponding fixed device of cultivation platform
In；
(2) after culture cup rotation mixes sample and culture medium, cultivation platform and culture cup carry out planetary motion, and sample is being cultivated
It is moved in cup, extracts the microorganism in sample；
(3) sample in culture cup being held up with strainer, the culture solution that enrichment facility is drawn in culture cup is rinsed sample,
Enrichment facility is passed through the culture solution in filtrated air concentration culture cup after strainer decline；Repeat this step；
(4) it shifts and collects concentrate.
2. automated food microorganism detection pre-treating method according to claim 1, which is characterized in that after being put into sample
Culture cup is independent sealed.
3. automated food microorganism detection pre-treating method according to claim 1, which is characterized in that the method is being set
It is equipped in the confined space of exhaust gas discharge conduit and carries out.
4. automated food microorganism detection pre-treating method according to claim 3, the exhaust gas discharge conduit is being cultivated
It is connected in concentration process with each culture cup.
5. according to claim 1 or the 3 automated food microorganism detection pre-treating methods, which is characterized in that along the training
It supports cup surrounding and is provided with thermostat.
6. automated food microorganism detection pre-treating method according to claim 5, which is characterized in that step (2) institute
The temperature for stating thermostat setting is 37 DEG C；The temperature of step (3) the thermostat setting is 45 DEG C.
7. automated food microorganism detection pre-treating method according to claim 1 or claim 2, which is characterized in that the culture
Cup inner wall is provided with protrusion.
8. automated food microorganism detection pre-treating method according to claim 1, which is characterized in that concentrate has shifted
Bi Hou, culture cup are destroyed as medical disposable material.
9. automated food microorganism detection pre-treating method according to claim 1, which is characterized in that before detection pre-treatment
After further include cleaning gas and/or force sterilizing program.
10. a kind of automated food microorganism detection pretreating device, which is characterized in that including closed main body, and be set to
The closed master is intracorporal can closed independent culture cup, cultivation platform, enrichment facility, thermostat and safety guarantee pressure dress
It sets；The cultivation platform can be equipped with the fixed device of culture cup, institute relative to the closed horizontal rotation of main body, cultivation platform
Culture cup is stated with the fixed horizontal rotation of device；Strainer is provided in culture cup, for lifting sample；The enrichment facility is in strainer
Imbibition flushing is carried out to sample when rising, enrichment facility is passed through filtrated air and is concentrated after strainer decline；The thermostat
Culture and the thickening temperature of needs are provided for the culture cup；The safety guarantee forcing device includes ultraviolet radiator and sterile logical
Device of air is carried out forcing sterilizing before and after booting by process control and is passed through filtrated air operation.
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| WO2022052423A1 (en) * | 2020-09-14 | 2022-03-17 | 倍仪昇智能科技(苏州)有限公司 | Pretreatment system and pretreatment method for microorganism detection |
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