CN205420407U - Witnessed inspections total bacteria count's detect reagent box - Google Patents
Witnessed inspections total bacteria count's detect reagent box Download PDFInfo
- Publication number
- CN205420407U CN205420407U CN201521049595.6U CN201521049595U CN205420407U CN 205420407 U CN205420407 U CN 205420407U CN 201521049595 U CN201521049595 U CN 201521049595U CN 205420407 U CN205420407 U CN 205420407U
- Authority
- CN
- China
- Prior art keywords
- reagent bottle
- groove
- reference point
- reaction cup
- luciferase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The utility model relates to a witnessed inspections total bacteria count's detect reagent box, this kit including the box inner support, the kit still including reagent bottle, the reagent bottle that holds the two phosphatases of three adenosine phosphate, the reagent bottle that holds the two phosphatases of three adenosine phosphate that hold the luciferase, hold and be used for diluting the two phosphatase buffer reagent bottles of three adenosine phosphate of the two phosphatases of three adenosine phosphate, the reagent bottle that holds surface -activeagent, the reagent bottle that holds the ATP standard substance and syringe needle formula response filter ware, the setting of box inner support is used for placing the standing groove of syringe needle formula response filter ware to and be used for placing the standing groove of reagent bottle. This kit has to carry and preserve and makes things convenient for safety, easy operation, and the result shows advantages such as rapid.
Description
Technical field
This utility model is the detection kit about a kind of Site Detection total number of bacteria.
Background technology
ATP detection technique of fluorescence is to react luminous by ATP in organism (adenosine triphosphate) under the effect of ion with luciferase, fluorescein, reacts biological pollution degree by the intensity of light.
At present, microorganism detection mainly uses culture method, takes back laboratory cultures and obtain total number of bacteria in 48 hours after collection in worksite sample.There is a lot of problem in this method: as 48 hours go out result shortage actual effect, especially when Security ensuring of important activities and burst fire-disaster, it is impossible to quickly obtain result.It addition, sample is cultivated after taking laboratory to, the bacterial death that some drags are relatively low can be made, it is impossible to the microorganism pollution level that real reaction is on-the-spot.
ATP detection technique of fluorescence can judge biological pollution degree in one minute, relatively culture of microorganism is the most effective, but current ATP fluoroscopic examination is only able to display relative light unit (RLU), relative light unit a lot of weak points in actual applications, this is mainly manifested in: the relative light unit (RLU) of the instrument of (1) different sensitivity and reagent detection same concentration sample is different, it is impossible to obtain unified result；(2) interference of non-bacterial ATP cannot be got rid of when detecting total number of bacteria, therefore cannot detect the clump count in natural environment at the scene；(3) owing to there being ion to participate in reaction when ATP bioluminescence detects, the ion in nature can excite or the fluorescence of cancellation reaction, makes relative light unit deviation.
Additionally, ATP detection technique of fluorescence needs plurality of reagents, multiple containers need to be used, carry and use inconvenient, and in view of the polytropy of Site Detection total number of bacteria environment, during operator's practical operation, the as easy as rolling off a log use order by reagent mixes up, and causes and cannot quickly obtain result.
In view of this, this utility model people relies on experience and the practice being engaged in relevant industries for many years, proposes the detection kit of a kind of Site Detection total number of bacteria, to overcome the defect of prior art.
Utility model content
The purpose of this utility model is to provide the detection kit of a kind of Site Detection total number of bacteria, this test kit to have to carry and store convenient and safe, simple to operate, and result shows the advantage such as rapidly.
For achieving the above object, this utility model provides the detection kit of a kind of Site Detection total number of bacteria, this test kit includes box inner support, wherein: described test kit also includes holding the reagent bottle of luciferase, holds the reagent bottle of buffer for diluting luciferase, holds the reagent bottle of apyrase, holds the reagent bottle of buffer for diluting apyrase, holds the reagent bottle of surfactant, the reagent bottle holding ATP standard substance and needle-based filter response；
Described box inner support is provided for placing the placing trough of needle-based filter response, and for placing the placing trough of reagent bottle；
Described needle-based filter response includes reaction cup and syringe-driven filter, this reaction cup top arranges well, bottom arranges and arranges airtight mouth stuffed on outage and outage, arranging the filter membrane of aperture 0.45 ± 0.2 μm in reaction cup, the microorganism being divided into filter membrane top inside reaction cup is retained space and the space of filter membrane bottom by filter membrane；
Described syringe-driven filter is mounted on above the well of reaction cup, and the filtrate (liquid of syringe-driven filter retains space with the microorganism on reaction cup filter membrane top and connects.
The detection kit of this utility model Site Detection total number of bacteria has carries, stores convenient and safe, simple to operate, and result shows the advantage such as rapidly.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, to place the groove of reagent bottle holding apyrase as reference point, the reagent bottle holding the buffer for diluting apyrase is positioned in the groove above described reference point, and the reagent bottle holding surfactant is positioned in the groove below described reference point；
Hold the reagent bottle of luciferase and be positioned on the left of described reference point in the groove of top, described in hold on the left of the reagent bottle diluting luciferase buffer is positioned over described reference point in the groove of lower section；
The reagent bottle holding ATP standard substance is positioned on the right side of described reference point in the groove of top, and described needle-based filter response is positioned in the groove of this reference point lower right-hand side.
Preferably, to place the groove of reagent bottle holding apyrase as reference point, the reagent bottle holding the buffer for diluting apyrase is positioned in the groove directly over described reference point, and the reagent bottle holding surfactant is positioned in the groove immediately below described reference point；
The reagent bottle holding luciferase is positioned in the groove above on the left of described reference point, holds in the groove below on the left of the reagent bottle diluting luciferase buffer is positioned over described reference point；And the groove below described left side is positioned at the underface of the groove above described left side；
The reagent bottle holding ATP standard substance is positioned on the right side of described reference point in the groove of top, and described needle-based filter response is positioned in the groove of described reference point lower right-hand side, and the groove of described lower right-hand side is positioned at the underface of the groove above described right side.
By parts each in as above position relationship places this test kit, tester is made can progressively to use the parts in test kit according to the detection method provided, it is not easy to cause confusion during use.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, described luciferase is the luciferase of lyophilizing, and described apyrase is the apyrase of lyophilizing；Described surfactant is quaternary cationics；Described is PBS solution for diluting the buffer of luciferase；Described is PBS solution for diluting the buffer of apyrase.Above reagent all can business enough obtain, wherein, apyrase comprises a phosphatase and diphosphatase, and two kinds of enzymes jointly act on and adenosine triphosphate can be degraded to adenosine monophosphate, can effectively get rid of the interference to Bacteria Detection of non-bacterial ATP, its degradation efficiency can reach more than 90%.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, it is tightly connected by chimeric or engagement thread between filtrate (liquid and the well of reaction cup of described syringe-driven filter.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, described syringe-driven filter is the syringe-driven filter of aperture 5 ± 0.2 μm.The needle-based filter membrane using this specification can be easy to filter the bulky grain in testing sample liquid and impurity.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, described reaction cup volume is 1mL～100mL.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, described reaction cup is to be applicable to the transparent reaction cup that fluorescence detector carries out detecting.Use transparent reaction cup to be directly placed in fluorescence detector by reacted liquid to be measured, decrease the contaminated possibility of liquid to be measured.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, described well is arranged on reaction cup crown center position, and described outage is arranged on centre position bottom reaction cup.Use this set to be more beneficial for the addition etc. of reagent during ATP bioluminescence method detects to operate.
According to specific embodiments of the present utility model, in the detection kit of described Site Detection total number of bacteria, the filter membrane in described reaction cup is the position of close bottom in being fixedly installed on reaction cup.
The surfactant held in this utility model reagent bottle is quaternary surfactant, it can press down the inhibitor of apyrase, thus suppress the Degradation of adenosine triphosphate, and can quickly make the polysaccharide structures disintegrate of bacteria cell wall, effectively extract the adenosine triphosphate in bacterial cell.
The luciferase of the lyophilizing held in this utility model reagent bottle can be the recombinant type luciferase of lyophilizing, there is preferable heat stability, it can business enough obtain, and reacts luminescence with the described adenosine triphosphate dissolving afterwards for diluting luciferase buffer and extracting from bacterial cell and obtain relative light unit during use.
The liquid ATP sterling that ATP standard substance are known quantity held in this utility model reagent bottle, it is commercially available, calculates the content of ATP in antibacterial by the ATP standard substance of known quantity, and calculates antibacterial number.Use holds ATP standard substance reagent bottle can get rid of the external environment interference (such as ion concentration, reaction temperature etc.) to experiment.
Test kit of the present utility model, when for the detection microorganism of ATP bioluminescence method, can use following operation to carry out:
(1) the airtight mouth stuffed on outage bottom described reaction cup is opened, asepsis injector is used to take certain volume measuring samples liquid (such as tap water or food leaching liquid), add the adding mouth of syringe-driven filter, testing sample liquid passes through syringe-driven filter, it is filtered to remove big granule and impurity, in entrance reaction cup, flowing through the filter membrane in reaction cup, the antibacterial in testing sample liquid is by membrane retention；
(2) apyrase after using the dilution of described apyrase buffer adds in needle-based filter response, stands 10 minutes；
(3) liquid of evacuation needle hair style filter response；
(4) in the liquid of needle-based filter response, add efficient surfactant, extract the adenosine triphosphate in antibacterial；
(5) in the liquid of needle-based filter response, the luciferase after using buffer dilution is added；
(6) detection relative light unit RLU1；
(7) add ATP standard substance and again detect relative light unit RLU2, by the ATP standard substance of known quantity, by RLU1/RLU2=X/A, (X is ATP content in sample, A is the amount of ATP standard substance used), calculate the content of ATP in antibacterial, and calculate antibacterial number.
In sum, this utility model provides the detection kit of a kind of Site Detection total number of bacteria, and this test kit has and carries, stores convenient and safe, simple to operate, and result show the advantages such as rapid.
Accompanying drawing explanation
The following drawings is only intended to, in schematically illustrating this utility model and explaining, not limit scope of the present utility model, wherein:
Fig. 1 is the structural representation of the detection kit of this utility model Site Detection total number of bacteria；In figure, label has a following meaning:
100: test kit；101: box inner support；102～107: groove；108: place the groove of needle-based filter response；
Fig. 2 is the result schematic diagram of this utility model needle-based filter response, and in figure, label has a following meaning:
201: sample introduction end；202: syringe-driven filter；203: well；204: reaction cup；205: filter membrane；206: outage；207: airtight mouth stuffed.
Detailed description of the invention
In order to be more clearly understood from technical characteristic of the present utility model, purpose and effect, now comparison accompanying drawing illustrates detailed description of the invention of the present utility model.
As shown in Figure 1, this utility model provides the detection kit 100 of a kind of Site Detection total number of bacteria, this test kit 100 includes box inner support 101, wherein, described test kit 100 also includes holding the reagent bottle of the luciferase of lyophilizing, holds the reagent bottle of buffer for diluting luciferase, holds the reagent bottle of the apyrase of lyophilizing, holds the reagent bottle of buffer for diluting apyrase, holds the reagent bottle of surfactant, the reagent bottle holding ATP standard substance and needle-based filter response；
Described box inner support 101 is provided for placing the groove 108 of described needle-based filter response, and for placing the groove of reagent bottle；
Described needle-based filter response is as shown in Figure 2, it includes reaction cup 204 and syringe-driven filter 202, this syringe-driven filter filter sizes is 5 ± 0.2 μm, this reaction cup 204 top arranges well 203, bottom arranges and arranges airtight mouth stuffed 207 on outage 206 and outage, arranging the filter membrane 205 that aperture is 0.45 ± 0.2 μm in reaction cup 204, internal for reaction cup 204 microorganism being divided into filter membrane top is retained space and the space of filter membrane bottom by filter membrane 205；
Described syringe-driven filter 202 is mounted on above the well 203 of reaction cup 204, and the filtrate (liquid of syringe-driven filter retains space with the microorganism on reaction cup filter membrane top and connects.
To place the groove 104 of reagent bottle holding apyrase as reference point, the reagent bottle holding the buffer for diluting apyrase is positioned in the groove 105 directly over described reference point, and the reagent bottle holding surfactant is positioned in the groove 106 immediately below described reference point；
The reagent bottle holding luciferase is positioned in the groove 102 above on the left of described reference point, holds in the groove 103 below on the left of the reagent bottle diluting luciferase buffer is positioned over described reference point；And the groove 103 below described left side is positioned at the underface of the groove 102 above described left side；
The reagent bottle holding ATP standard substance is positioned in the groove 107 above on the right side of described reference point, described needle-based filter response is positioned in the groove 108 of described reference point lower right-hand side, and the groove 108 of described lower right-hand side is positioned at the underface of the groove 107 above described right side.
During use, the sample introduction end 201 of needle-based filter response is connected on syringe by the liquid sample after drawing 10mL pre-treatment with 10mL syringe, after pressurization, sample liquid passes through the cavity of reaction cup and discharges from discharge outlet 206, and in sample, antibacterial is dammed at 0.45 μm filter membrane 205.
nullAfter the apyrase of the buffer in the reagent bottle that will be positioned in groove 105 and the lyophilizing being positioned in groove 104 in reagent bottle dissolves，50 μ l are taken with pipettor，Add from the well 203 of needle-based filter response，Stand 10 minutes，It is made to empty after fully reacting with the thing that dams on filter membrane 205，The efficient surfactant that 50 μ l are positioned in groove 106 in reagent bottle is taken with pipettor，Add from the well 203 of needle-based filter response and vibrate，Being used in the luciferase of the lyophilizing in the reagent bottle that will be positioned in groove 102 reagent bottle in being positioned over groove 103 dilutes after luciferase buffer dissolves，400 μ l are taken with pipettor，Add from the well 203 of needle-based filter response，Put into the detection of ATP fluorescence detector and obtain relative light unit RLU1，Pipette the ATP standard substance that 10 μ l are positioned in the reagent bottle in groove 107 again，Add from the well 203 of needle-based filter response，Detection obtains relative light unit RLU2 again，By the ATP standard substance of known quantity, by RLU1/RLU2=X/A, (X is ATP content in sample，A is the amount of the ATP standard substance used)，Calculate the content of ATP in antibacterial，And calculate antibacterial number.
In sum, the detection kit of this utility model provided Site Detection total number of bacteria have carry, store convenient and safe, simple to operate, result shows the advantages such as rapid, and the application applying this test kit is simple and convenient, the detection of bacteria total amount in sample can be completed at short notice, substantially reduce the time of Bacteria Detection, can be widely used for food safety, medical and health organization, disinfection of epidemic focus effect, laboratory, pharmacy, the on-the-spot microorganism of food processing link are quickly examined.
The foregoing is only the schematic detailed description of the invention of utility model, be not limited to scope of the present utility model.Any those skilled in the art, equivalent variations done on the premise of without departing from design of the present utility model and principle and amendment, the scope of this utility model protection all should be belonged to.
Claims (10)
1. the detection kit of a Site Detection total number of bacteria, this test kit includes box inner support, it is characterised in that: described test kit also includes holding the reagent bottle of luciferase, holds the reagent bottle of buffer for diluting luciferase, holds the reagent bottle of apyrase, holds the reagent bottle of buffer for diluting apyrase, holds the reagent bottle of surfactant, the reagent bottle holding ATP standard substance and needle-based filter response；
Described box inner support is provided for placing the placing trough of needle-based filter response, and for placing the placing trough of reagent bottle；
Described needle-based filter response includes reaction cup and syringe-driven filter, this reaction cup top arranges well, bottom arranges and arranges airtight mouth stuffed on outage and outage, arranging the filter membrane of aperture 0.45 ± 0.2 μm in reaction cup, the microorganism being divided into filter membrane top inside reaction cup is retained space and the space of filter membrane bottom by filter membrane；
Described syringe-driven filter is mounted on above the well of reaction cup, and the filtrate (liquid of syringe-driven filter retains space with the microorganism on reaction cup filter membrane top and connects.
The detection kit of Site Detection total number of bacteria the most according to claim 1, it is characterized in that: to place the groove of reagent bottle holding apyrase as reference point, the reagent bottle holding the buffer for diluting apyrase is positioned in the groove above described reference point, and the reagent bottle holding surfactant is positioned in the groove below described reference point；
Hold the reagent bottle of luciferase and be positioned on the left of described reference point in the groove of top, described in hold on the left of the reagent bottle diluting luciferase buffer is positioned over described reference point in the groove of lower section；
The reagent bottle holding ATP standard substance is positioned on the right side of described reference point in the groove of top, and described needle-based filter response is positioned in the groove of this reference point lower right-hand side.
The detection kit of Site Detection total number of bacteria the most according to claim 2, it is characterized in that: to place the groove of reagent bottle holding apyrase as reference point, the reagent bottle holding the buffer for diluting apyrase is positioned in the groove directly over described reference point, and the reagent bottle holding surfactant is positioned in the groove immediately below described reference point；
The reagent bottle holding luciferase is positioned in the groove above on the left of described reference point, holds in the groove below on the left of the reagent bottle diluting luciferase buffer is positioned over described reference point；And the groove below described left side is positioned at the underface of the groove above described left side；
The reagent bottle holding ATP standard substance is positioned on the right side of described reference point in the groove of top, and described needle-based filter response is positioned in the groove of described reference point lower right-hand side, and the groove of described lower right-hand side is positioned at the underface of the groove above described right side.
4. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: described luciferase is the luciferase of lyophilizing, and described apyrase is the apyrase of lyophilizing；Described surfactant is quaternary cationics；Described is PBS solution for diluting the buffer of luciferase；Described is PBS solution for diluting the buffer of apyrase.
5. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: it is tightly connected by chimeric or engagement thread between filtrate (liquid and the well of reaction cup of described syringe-driven filter.
6. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: described syringe-driven filter is the syringe-driven filter of aperture 5 ± 0.2 μm.
7. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: described reaction cup volume is 1mL～100mL.
8. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that described reaction cup is to be applicable to the transparent reaction cup that fluorescence detector carries out detecting.
9. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: described well is arranged on reaction cup crown center position, and described outage is arranged on centre position bottom reaction cup.
10. according to the detection kit of the Site Detection total number of bacteria according to any one of claims 1 to 3, it is characterised in that: the filter membrane in described reaction cup is the position of close bottom in being fixedly installed on reaction cup.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201521049595.6U CN205420407U (en) | 2015-12-16 | 2015-12-16 | Witnessed inspections total bacteria count's detect reagent box |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201521049595.6U CN205420407U (en) | 2015-12-16 | 2015-12-16 | Witnessed inspections total bacteria count's detect reagent box |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN205420407U true CN205420407U (en) | 2016-08-03 |
Family
ID=56536900
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201521049595.6U Active CN205420407U (en) | 2015-12-16 | 2015-12-16 | Witnessed inspections total bacteria count's detect reagent box |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN205420407U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113652467A (en) * | 2021-09-01 | 2021-11-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Method and kit for rapidly determining number of oral microorganisms and application of kit |
-
2015
- 2015-12-16 CN CN201521049595.6U patent/CN205420407U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113652467A (en) * | 2021-09-01 | 2021-11-16 | 上海纳米技术及应用国家工程研究中心有限公司 | Method and kit for rapidly determining number of oral microorganisms and application of kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wilson et al. | Quantitative and qualitative assessment methods for biofilm growth: a mini-review | |
| KR200474522Y1 (en) | Full-enclosed filtration-culture ampoule system for sterility testing | |
| JP5591747B2 (en) | Luminescence measuring device and microorganism counting device | |
| CN201476980U (en) | Device for sampling and testing | |
| CN101464412B (en) | Biologic sensor for optical fast detection of surface cleanliness and its test method | |
| AU2016217678B2 (en) | System, devices and methods using an integrated sphere light collector | |
| CN102203588A (en) | Methods for separation, characterization and/or identification of microorganisms using spectroscopy | |
| JPH11514849A (en) | Inspection apparatus, system and method for detecting an inspection sample | |
| US8183052B2 (en) | Methods and apparatus for sterility testing | |
| CN205347422U (en) | Little biological detection device | |
| RU2517618C2 (en) | Method and system for determining quality of cultivated cells | |
| JPH04500307A (en) | Methods for detecting biological activity | |
| CN101113985B (en) | Sanitary status on-site rapid detection device and detecting method | |
| CN101918582A (en) | Utilize physical change in the blood and chemical transformation to detect microorganism in the blood | |
| Yu et al. | Rapid detection and enumeration of total bacteria in drinking water and tea beverages using a laboratory-built high-sensitivity flow cytometer | |
| CN102600918B (en) | Microfluidic chip for blood analysis and method for using microfluidic chip | |
| CN101130808A (en) | Fast-culturing susceptibility and micro-detection method for mycobacteria and device thereof | |
| CN205420407U (en) | Witnessed inspections total bacteria count's detect reagent box | |
| CN105203510B (en) | A kind of microorganism in food rapid detection method | |
| JP7457772B2 (en) | Luminescence measuring device and luminescence measuring method | |
| CN204101457U (en) | A kind of reaction unit detecting microorganism for ATP bioluminescence method | |
| CN103776807A (en) | Method for detecting residual bacteria in flushed water of CIP (cleaning in place) cleaning system | |
| US5416002A (en) | Near-real-time microbial monitor | |
| Chen et al. | Real-time monitoring for bioaerosols—flow cytometry | |
| CN104673664A (en) | Blood sample culture device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |