CN107779393A - Microorganism sampler - Google Patents
Microorganism sampler Download PDFInfo
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- CN107779393A CN107779393A CN201711189251.9A CN201711189251A CN107779393A CN 107779393 A CN107779393 A CN 107779393A CN 201711189251 A CN201711189251 A CN 201711189251A CN 107779393 A CN107779393 A CN 107779393A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/24—Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
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Abstract
The present invention relates to scientific research and medical detection technology, disclose a kind of microorganism sampler, including storage bottle, it is homogenized ball, injector, bottle cap, the storage bottle bottom is provided with sampling cavity, storage bottle top outer is provided with screw thread, homogenate ball is provided with inside storage bottle, it is connected at the top of the storage bottle with injector, sample cavity is respectively equipped with inside the injector, limit chamber, connecting cavity, screw thread is equipped with the outside of sample cavity and on the inside of connecting cavity, baffler is provided between sample cavity and limit chamber, baffler is provided with groove, the bottle cap is connected with injector, closure inside is provided with projection, be nested sealing ring on the outside of projection.The microorganism sampler, by setting the fixation useful load of sample cavity, it is easy to user to accurately hold the collection capacity of sample, while the good seal performance of the present apparatus, effectively prevent sample leakage, is easy to preserve or transport for a long time.
Description
Technical field
The present invention relates to scientific research and medical detection technology, specially a kind of microorganism sampler.
Background technology
Microorganism all plays a very important role in the chemical balance in biosphere and the ecological balance, such as right in recent years
The research of gut flora is awfully hot, and it, which is studied, is mainly focused on digestion of the gut flora to host and to the function such as immune and disease-resistant
Material impact.Past research have shown that the intestinal microflora of people has significant individual difference, can by Molecular tools
The molecular labeling of flora is shared for search of health enteron aisle so as to make more accurate diagnosis to intestines problem；Ground in food
In studying carefully, due to the appearance of drug-fast bacteria in food, food is polluted by fecal bacteria, drinking-water, food and feed additive and veterinary drug
The reason such as influence, application of the probiotics in gastrointestinal bacterial flora disorder is prevented and treated to gut flora is remained, makes people couple
The Tiny ecosystem research of gut flora is increasingly paid close attention to.Research for edaphon Bacterial community equally has very big exploitation
Potentiality, study Soil Microorganism composition and dynamic change specifying information while, also the diagnosis to practical problem, make
The farming operations such as thing growth tracking and monitoring, rhizosphere effect and straw-returning all have most important theories and practical value.In lake
In the ecosystem, material is circulated microorganism and energy flow plays huge effect, for the work of effective microorganisms
With must just analyze its group, abundant microbial diversity information is obtained.
The technology hand that analysis to excrement, soil or lake silt microbial diversity is scientific research and medical field is commonly used
Section.Micro- life of human intestine's flora, soil ecosystem and lake ecosystem can be accurately understood by this detection
Thing Bacterial community.Carrying out this detection needs to use the sampler of convenient science to be sampled, but existing sampler
Seldom there is the function of quantitative sampling, it is difficult the collection capacity for holding sample to cause user, while does not have fully dispersed solid-state
The function of sample, and it is solid-state or semisolid that the sample of Institute of Micro-biology's parasitism is most of, and then cause extraction sample can not be with guarantor
Liquid storage it is fully molten and, and then influence microbiological analysis result, and existing sampler sealing property is not good enough, easily causes
Leakage, the problem of being not easy to preserve for a long time.
The content of the invention
(1) technical problem solved
In view of the shortcomings of the prior art, the invention provides a kind of microorganism sampler, possess quantitative sampling, make sample
With preserve liquid fully it is molten and, be easy to sample, and the advantages that sampler favorable sealing property, solve used microorganism and take
Sample device without quantitative sampling, without scattered sample medium and poor sealing performance, not readily transportable or what is preserved for a long time asks
Topic.
(2) technical scheme
To achieve the above object, the present invention provides following technical scheme：A kind of microorganism sampler, including storage bottle,
Ball, injector, bottle cap are homogenized, the storage bottle bottom is provided with sampling cavity, and storage bottle top outer is provided with screw thread, inside storage bottle
Provided with homogenate ball, it is connected at the top of the storage bottle with injector, sample cavity, limit chamber, company is respectively equipped with inside the injector
Chamber is connect, screw thread is equipped with the inside of sample cavity outside and connecting cavity, baffler is provided between sample cavity and limit chamber, baffler is provided with
Groove, the bottle cap are connected with injector, and closure inside is provided with projection, and be nested sealing ring on the outside of projection.
Preferably, the sample cavity of the injector and connecting cavity internal diameter are unequal, and connecting cavity internal diameter is straight more than sample cavity
Footpath.
Preferably, the sample cavity is equal with limit chamber internal diameter, internal diameter and limit chamber internal diameter phase at the storage bottle port
Deng.
Preferably, the circumferentially Array Design of the groove on the baffler.
Preferably, the homogenate ball is spherical, and can place two to four homogenate balls in storage bottle.
Preferably, the projection is cylinder, is fixedly connected on closure inside middle, and when bottle cap and sample cavity outside
When threaded connection is tightened, sample cavity port contacts with sealing ring extruding, and projection contacts just with baffler.
Preferably, the sampling cavity is conical design, and sampling cavity base material thickness bottom is smaller than storage bottle.
(3) beneficial effect
Compared with prior art, the invention provides a kind of microorganism sampler, possesses following beneficial effect：
The microorganism sampler, can be to solid-states such as excrement, soil or silts by setting the fixation useful load of sample cavity
Or semisolid sample carries out quantitative sampling, it is easy to user to accurately hold the collection capacity of sample, while by setting baffler, leading to
Groove and homogenate ball sample is sufficiently disperseed, make sample with preserve liquid carry out sufficiently it is molten and, stabilization sample in microorganism
Composition, this sampler is more preferable to the integrity protection effect of sample, do not influence follow-up DNA detection, while the present apparatus
Good seal performance, sample leakage is effectively prevent, be easy to preserve or transport for a long time.
Brief description of the drawings
Accompanying drawing is used for providing a further understanding of the present invention, and a part for constitution instruction, the reality with the present invention
Apply example to be used to explain the present invention together, be not construed as limiting the invention, in the accompanying drawings：
Fig. 1 is exploded perspective view of the present invention；
Fig. 2 is overall structure diagram of the present invention；
Fig. 3 is entirety sectional view of the present invention；
Fig. 4 is injector structure schematic diagram of the present invention；
Fig. 5 is injector sectional view of the present invention；
Fig. 6 is bottle cap sectional view of the present invention；
Fig. 7 is the DNA extraction results (agarose gel electrophoresis testing result) of the invention with sample in common centrifuge tube；
Fig. 8 is the DNA sample result data table of the invention with common centrifuge tube.
In figure：1- storage bottles, 11- sampling cavities, 2- homogenate ball, 3- injectors, 31- sample cavitys, 32- bafflers, 33- lead to
Groove, 34- connecting cavities, 35- limit chambers, 4- bottle caps, 5- sealing rings, 6- projections.
Embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation describes, it is clear that described embodiment is only part of the embodiment of the present invention, rather than whole embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art are obtained every other under the premise of creative work is not made
Embodiment, belong to the scope of protection of the invention.
Accompanying drawing is referred to, the present invention provides a kind of technical scheme：A kind of microorganism sampler, including storage bottle 1, homogenate
Ball 2, injector 3, bottle cap 4, the bottom of storage bottle 1 are provided with sampling cavity 11, and the top outer of storage bottle 1 is provided with screw thread, inside storage bottle 1
Provided with homogenate ball 2, the top of storage bottle 1 is connected with injector 3, and sample cavity 31, limit chamber are respectively equipped with inside injector 3
35th, connecting cavity 34, the outside of sample cavity 31 and the inner side of connecting cavity 34 are equipped with screw thread, and resistance is provided between sample cavity 31 and limit chamber 35
Dividing plate 32, baffler 32 are provided with groove 33, and the bottle cap 4 is connected with injector 3, and the inside of bottle cap 4 is provided with projection 6, the outside of projection 6
Be nested sealing ring 5.
The sample cavity 31 of injector 3 and the internal diameter of connecting cavity 34 are unequal, and the internal diameter of connecting cavity 34 is more than the diameter of sample cavity 31,
It is easy to connecting cavity 34 to accommodate the port of storage bottle 1.
Sample cavity 31 is equal with the internal diameter of limit chamber 35, and internal diameter is equal with the internal diameter of limit chamber 35 at the port of storage bottle 1, is easy to store up
When depositing bottle 1 and being threadedly coupled with connecting cavity 34, limit chamber 35 carries out spacing and screens to the port of storage bottle 1, and then ensures storage bottle 1
It is fixedly connected with connecting cavity 34, and the end face of limit chamber 35 is extruded by the port of storage bottle 1, it is therefore prevented that in storage bottle 1
Sample stores thing and leaked from storage bottle 1 and the junction of connecting cavity 34.
Groove 33 on baffler 32 circumferentially Array Design, is easy to microorganism to be entered by groove 33 in storage bottle 1, simultaneously
When the groove 33 that hard micro-biological samples are designed by circumference array, play a part of scattered hard micro-biological samples.
Ball 2 is homogenized to be spherical, and two to four homogenate balls 2 can be placed and arrived in storage bottle 1, the homogenate matter of ball 2 is hard, chemical property
Torpescence, and water, acidity or alkaline environment are insoluble in, so after solid-state or semisolid dispersate sample enter storage bottle 1,
By rocking storage bottle 1, homogenate ball 2, buffer solution and sample in storage bottle 1 mutually collide, and further disperse microorganism, are easy to
Subsequently through the microorganism in pipettor extraction sample homogenate.
Projection 6 is toroidal, is fixedly connected at the bosom of bottle cap 4, and when bottle cap 4 and the outside screw of sample cavity 31 connect
Connect when tightening, the port of sample cavity 31 contacts with the extruding of sealing ring 5, and projection 6 contacts just with baffler 32, is easy to place in sample
After in sample cavity 31, during being fixedly connected by bottle cap 4 and the screw thread of injector 3, the projection 6 inside bottle cap 4 extrudes
Sample enters in storage bottle 1 from the groove 33 of baffler 32, and the last port of sample cavity 31 extrudes with sealing ring 5, ensure that bottle cap 4
With the sealing between injector 3, and then avoid sample from leaking, while shifted beneficial to sample.
Sampling cavity 11 is conical design, and the base material thickness bottom of sampling cavity 11 is smaller than storage bottle 1, is easy to subsequently through shifting
Liquid device enters the detection that sampling cavity 11 extracts the further microorganism of progress of sample homogenate in storage bottle 1.
Shown in Fig. 7, the excrement of eight volunteers is sampled using the present apparatus and examines DNA, is reused now on the market
Common centrifuge tube sampled on an equal basis, by contrast, this sampler is more preferable to the integrity protection effect of sample, does not influence
Follow-up DNA detection, and common centrifuge tube is poor to the integrity protection effect of sample, or even the DNA destroyed in sample, sternly
Ghost image rings follow-up DNA detection.
When in use, because the height and diameter of sample cavity 31 are set, so further can be closed to sample sampling amount
Suitable restriction, first sample is moved into sample cavity 31, until sample is fully filled with sample cavity 31, then strike off the top opening of sample cavity 31
Sample, redundance is removed, then the screw thread of bottle cap 4 and the outside of sample cavity 31 is connected, as the rotation of bottle cap 4 is twisted
Tightly, the projection 6 inside bottle cap 4 is extruded sample, and sample enters in storage bottle 1 by groove 33, passes through circumference array point
The groove 33 of cloth has carried out first scattered to sample, bottle cap 4 coordinates with injector 3 tighten after, then the present apparatus rocked
And shake, mutually collided by the homogenate ball 2 in storage bottle 1, preservation liquid and sample, further disperse sample, be easy to pass through shifting
The sampling cavity 11 that liquid device pipette tips enter the bottom of storage bottle 1 draws the analysis that sample homogenate in storage bottle 1 carries out further microorganism.
Embodiment 1
One：Sample source and packet preserving type：
(1) the fresh excreta sample of 8 volunteers, every excrement are acquired according to the operating process of this microorganism sampler
Just at least 0.5g is sampled.
(2) with the common centrifuge tube collections of 5ml with the fresh excreta of crowd volunteer, band sealing.
Under two groups of sample normal temperature (25 DEG C) preserve 7 days after, carry out DNA extractions.
Two：Fecal sample processing method：
Genome DNA extraction is carried out to 2 parts of stool samples of above-mentioned 8 volunteers, processing procedure is as follows：
Prepare before experiment：E.Z.N.A.Stool DNA Kit 50T kits；1.5ml centrifuge tube × 2；2ml centrifuge tubes
×1；Absolute ethyl alcohol；Ice centrifuge tube box；Normal temperature centrifuges base.
Required instrument：Supercentrifuge；70 DEG C of water-baths；Vortex oscillator；Timer；Ice machine；Weigher；Liquid-transfering gun
1ml, 200 μ L, 20 μ L；100ml graduated cylinders.
Experimental procedure：
(1) add 80mL (96%-100%) absolute ethyl alcohols into DNA Wash Buffer with dilution.
(2) the dress 200mg glass beads in 2ml centrifuge tubes (providing for oneself).Precooling on ice.Sample is scraped using scraper
50-100mg。
(3) sample melt before rapidly plus 300 μ L Buffer SP1,10 μ L Proteinase K solution to from
In heart pipe.Whirlpool maximal rate concussion 5min is until fecal specimens substantially uniformity.
(4) 70 DEG C incubation, during need by be vortexed mix twice.13min is incubated altogether.Gram-positive bacteria needs to increase
90 DEG C, 5min.
(5) 2min is incubated on ice.
(6) 100 μ L Buffer SP2 are added.Shaking 30s by whirlpool makes sample mixing abundant.
(7) 5min is incubated on ice.
(8) 13000-20000g, which centrifuges 5min, makes excrement form graininess.
(9) carefully draw upper epidermis (to provide for oneself) into 1.5ml centrifuge tubes, it is ensured that avoid particle or fragment.
(10) plus 200 μ L HTR are into 1.5ml centrifuge tubes, and mixing, whirlpool concussion 10s are fully aspirated using 1ml pipettors.
(11) incubation at room temperature 2min.
(12) it is more than 13000g centrifugations 2min, it is therefore an objective to inhibitor is absorbed HTR into graininess.
(13) 250 μ L of supernatant are drawn into new 2ml pipes.
(14) 250 μ L BL Buffer are added, 250 μ l absolute ethyl alcohols are into lysate.It is fully mixed that 10s is shaken by whirlpool
Close sample.
(15) the overall sample in upper step is included into precipitation, is added in DNA posts and 2ml collecting pipes (offer) are provided.It is more than
13000g room temperatures centrifuge 1min.Discard liquid and bottom collection pipe.
(16) pillar is arranged in new 2ml collecting pipes (offer), adds 500 μ L HB Buffer.More than 13000g
Centrifuge 1min.Discard liquid and collecting pipe.
(17) pillar is arranged in new 2ml collecting pipes (offer).Add 750 μ l DNA Wash Buffer (dilutions
Cross), centrifuge 1min more than 13000g.Liquid is discarded, pillar is reinserted in the collecting pipe of sky.
(18) it is more than 13000g room temperatures centrifugation 2min, dries pillar.This step is very crucial, after removing ethanol in order to avoid disturbing
The step of.
(19) pillar is enclosed in a new 1.5ml pipe and (provided for oneself).Elution Buffer need 60-70 DEG C of preheating, add
200 μ l are directly in the matrix of bottom of the pillar.2min is infiltrated at room temperature.1min is centrifuged more than 13000g.
Three：Fecal sample DNA concentration, quality and Purity：
After the completion of DNA extractions：
(1) DNA concentration is detected：UseDsDNA BR assay kits.Examination is diluted with the buffer solution of kit
Agent, the μ L of sample 1 are added, are then used2.0 luminoscopes read concentration.
(2) DNA mass is detected：1 μ l DNA are taken, are detected respectively always using NanoDrop 2000C ultramicrospectrophotometers
Absorbances of the DNA at 260nm and 280nm two, obtains the concentration of STb gene and OD260/OD280 ratio, if the ratio exists
Between 1.6-1.8, illustrate that the DNA of sample purity is higher, no protein and RNA pollution；If ratio<1.6, illustrate there is protein
Pollution；If ratio between 1.8-2.0, illustrates there are RNA pollutions.
(3) DNA purity is detected：Agarose gel electrophoresis method.0.36g agaroses are weighed, are put into conical flask, add 60ml
TAE buffer solutions, preservative film sealing, put micro-wave oven and be heated to complete clear and take out to shake up, add EB, it is in glue to pour into groove solidification.
Point sample after DNA sample mixes with buffer, glue is run, band is observed under uviol lamp, referring particularly to Figure of description 7.
In summary, the microorganism sampler, by setting the fixation useful load of sample cavity, can to excrement, soil or
The solid-state or semisolid such as silt sample carries out quantitative sampling, is easy to user to accurately hold the collection capacity of sample, while by setting
Put baffler, groove and homogenate ball sample is sufficiently disperseed, make sample with preserve liquid carry out sufficiently it is molten and, stabilization sample
Composition of microorganism, the microorganism being more beneficial in subsequent extracted sample are further analyzed in this, while the present apparatus
Good seal performance, sample leakage is effectively prevent, be easy to preserve or transport for a long time, solve used microorganism sampler not
With quantitative sampling, without decentralized medium and poor sealing performance, not readily transportable or the problem of preserve for a long time.
It should be noted that herein, such as first and second or the like relational terms are used merely to a reality
Body or operation make a distinction with another entity or operation, and not necessarily require or imply and deposited between these entities or operation
In any this actual relation or order.Moreover, term " comprising ", "comprising" or its any other variant are intended to
Nonexcludability includes, so that process, method, article or equipment including a series of elements not only will including those
Element, but also the other element including being not expressly set out, or it is this process, method, article or equipment also to include
Intrinsic key element.In the absence of more restrictions, the key element limited by sentence "including a ...", it is not excluded that
Other identical element also be present in process, method, article or equipment including the key element.
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of changes, modification can be carried out to these embodiments, replace without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (7)
1. a kind of microorganism sampler, including storage bottle (1), homogenate ball (2), injector (3), bottle cap (4), its feature exist
In：Storage bottle (1) bottom is provided with sampling cavity (11), and storage bottle (1) top outer is provided with screw thread, set inside storage bottle (1)
There is homogenate ball (2), be connected at the top of the storage bottle (1) with injector (3), sample cavity is respectively equipped with inside the injector (3)
(31), limit chamber (35), connecting cavity (34), sample cavity (31) outside and connecting cavity (34) inner side are equipped with screw thread, sample cavity
(31) baffler (32) is provided between limit chamber (35), baffler (32) is provided with groove (33), the bottle cap (4) and injector
(3) connect, projection (6) is provided with inside bottle cap (4), be nested sealing ring (5) on the outside of projection (6).
2. microorganism sampler according to claim 1, it is characterised in that：The sample cavity (31) of the injector (3)
It is unequal with connecting cavity (34) internal diameter, and connecting cavity (34) internal diameter is more than sample cavity (31) diameter.
3. according to the microorganism sampler described in claim 1 and 2, it is characterised in that：The sample cavity (31) and limit chamber
(35) internal diameter is equal, and internal diameter is equal with limit chamber (35) internal diameter at storage bottle (1) port.
4. microorganism sampler according to claim 1, it is characterised in that：Groove (33) on the baffler (32)
Circumferentially Array Design.
5. microorganism sampler according to claim 1, it is characterised in that：The homogenate ball (2) is spherical, and can be put
It is interior to storage bottle (1) to put two to four homogenate balls (2).
6. microorganism sampler according to claim 1, it is characterised in that：The projection (6) is toroidal, fixed to connect
It is connected at bottle cap (4) bosom, and when bottle cap (4) is connected with sample cavity (31) outside screw and tightened, sample cavity (31) end
Mouth contacts with sealing ring (5) extruding, and projection (6) contacts just with baffler (32).
7. microorganism sampler according to claim 1, it is characterised in that：The sampling cavity (11) sets for cone
Meter, and sampling cavity (11) base material thickness bottom is smaller than storage bottle (1).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711189251.9A CN107779393A (en) | 2017-11-24 | 2017-11-24 | Microorganism sampler |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711189251.9A CN107779393A (en) | 2017-11-24 | 2017-11-24 | Microorganism sampler |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107779393A true CN107779393A (en) | 2018-03-09 |
Family
ID=61430747
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711189251.9A Pending CN107779393A (en) | 2017-11-24 | 2017-11-24 | Microorganism sampler |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110160819A (en) * | 2018-08-16 | 2019-08-23 | 嘉兴行健生物科技有限公司 | Fecal specimens are collected and extraction element |
-
2017
- 2017-11-24 CN CN201711189251.9A patent/CN107779393A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110160819A (en) * | 2018-08-16 | 2019-08-23 | 嘉兴行健生物科技有限公司 | Fecal specimens are collected and extraction element |
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