CN108913544A - Pathogenic microorganism efficiently concentrating device and its enrichment method in a kind of water environment - Google Patents

Pathogenic microorganism efficiently concentrating device and its enrichment method in a kind of water environment Download PDF

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Publication number
CN108913544A
CN108913544A CN201810870930.0A CN201810870930A CN108913544A CN 108913544 A CN108913544 A CN 108913544A CN 201810870930 A CN201810870930 A CN 201810870930A CN 108913544 A CN108913544 A CN 108913544A
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water
filter core
water sample
elution
control section
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尹静
王华然
杨忠委
李君文
金敏
杨栋
谌志强
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Environmental Medicine and Operational Medicine Institute of Military Medicine Institute of Academy of Military Sciences
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Environmental Medicine and Operational Medicine Institute of Military Medicine Institute of Academy of Military Sciences
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Publication of CN108913544A publication Critical patent/CN108913544A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/14Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes

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  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)

Abstract

The present invention relates to pathogenic microorganism efficiently concentrating devices in water environment, belong to water environment technical field of microbial detection.The enriching apparatus includes filtration fraction and control section, and filter core is placed in filter core barrel bore in filtration fraction and there are spaces between filter core and filter core barrel bore wall;Filter core sleeve top is provided with water sample water inlet;Filter core bottom end is provided with water sample water outlet;The inner wall that water sample water outlet passes through filter core sleeve extends to outside;Control section includes threeway, water sample filtering control section and elution control section;Water sample filters control section and has been sequentially communicated the first check valve and water sample filtering peristaltic pump according to water sample elution order;Elution control section has been sequentially communicated elution flow container, elution peristaltic pump and second one-way valve according to eluent inflow direction;Water sample water outlet, the first check valve are connected to second one-way valve by threeway.It realizes the efficiently concentrating to low concentration pathogenic microorganism in water environment, provides technical support for the pathophorous detection of Jie's water, monitoring.

Description

Pathogenic microorganism efficiently concentrating device and its enrichment method in a kind of water environment
Technical field
The invention belongs to the pathogenic microorganism examination technical fields in water environment, and in particular to the micro- life of cause of disease in a kind of water environment Object efficiently concentrating device and its enrichment method.
Background technique
All kinds of pathogenic microorganisms (including bacterium, protozoon, virus etc.) are that Jie's water is caused to spread the disease present in water environment The main reason for outburst, not only endangers population health, can also bring serious economic loss, or even generate serious bad society It will affect.The propagation of pathogenic microorganism Jie's water can frequently result in group and break out event, and global distribution is presented, wherein causing a disease Bacterium, helminth, virus etc. are main inducings.According to statistics, in world wide every year because caused by cause pathogeny imcrobe infection 5 years old with Lower children's diarrhae death toll is up to more than 130 ten thousand.
Jie's water propagate pathogenic microorganism generally have the characteristics that it is pathogenic strong, low amounts intake can lead to population infection, Its content in water environment is generally seldom simultaneously, and current common detection methods is caused to cannot achieve the accurate of pathogenic microorganism Detection, as a result, it is often necessary to by way of enrichment, by after being enriched with to the pathogenic microorganism in water sample with bulk mass It is able to satisfy the demand of detection.Currently, pathogenic microorganism enrichment common method has filtration method, flocculence, absorption method etc. in water environment, In practical applications can for different experiment purposes, water body environment and select different enrichment methods.
Filtration method is pathogenic microorganism enrichment method more common at present, generally based on microfiltration membranes, ultrafiltration membrane etc., but by It is limited in the ability that filter membrane is recycled, thus its cost is relatively high.Flocculation rule needs to come using a large amount of flocculant real The use of the purpose being now enriched with, a large amount of flocculants can cause certain potential hazard to environment.And absorption method is due to adsorbent material Regeneration it is comparatively laborious, influence being used repeatedly for adsorbent material.
Summary of the invention
In view of this, the purpose of the present invention is to provide pathogenic microorganism efficiently concentrating device and its richnesses in a kind of water environment Set method can be widely used in the enrichment of pathogenic microorganism in multiple water environments using described device.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme:
The present invention provides pathogenic microorganism efficiently concentrating devices in a kind of water environment, including filtration fraction and control unit Point:
The filtration fraction includes filter core and filter core sleeve;The filter core is placed in the filter core barrel bore and the filter core There are spaces between the internal chamber wall of the filter core sleeve;The top of the filter core sleeve is provided with water sample water inlet;The filter The bottom end of core is provided with water sample water outlet;The inner wall that the water sample water outlet passes through filter core sleeve extends to outside;
The control section includes threeway, water sample filtering control section and elution control section;The water sample filtering control Part has been sequentially communicated the first check valve and water sample filtering peristaltic pump according to water sample elution order;First check valve is opened Opening direction is opened along water sample filtering peristaltic pump direction;
The elution control section according to eluent inflow direction be sequentially communicated elution flow container 9, elution peristaltic pump and Second one-way valve;The opening direction of the second one-way valve is opened along water sample water outlet direction;
The second one-way valve of the water sample water outlet, the first check valve of water sample filtering control section and elution control section It is connected to by threeway.
Preferably, the material of the filter core sleeve includes stainless steel, glass, PVC or engineering plastics material.
Preferably, the aperture of the filter core includes 0.45~1 μm.
Preferably, the filter core includes stainless steel filter core, stud filter core or ceramic element.
Preferably, all connections are connected to using conduit in the control section.
Preferably, the material of the conduit includes emulsion tube, silicone tube, PVC hose etc..
Preferably, the enriching apparatus further includes cabinet;The filtration fraction and control section are located at the cabinets cavity In.
Preferably, the filtration fraction is removably attached in the inner cavity of cabinet.
Preferably, the method being removably secured includes screw, hasp or pipe collar.
Preferably, the water sample water inlet 4 extends to 10 outside of cabinet by conduit.
The present invention provides a kind of methods based on pathogenic microorganism in described device enriched water environment, including following step Suddenly:
1) flow velocity of water sample to be detected is controlled with water sample filtering with peristaltic pump, the flow velocity for setting water sample to be detected passes through Water sample water inlet enters the space between filter core sleeve and filter core, and the filtering of water sample is then realized through filtration core, filtered Water sample is discharged by water sample water outlet;
2) after water sample to be detected filters, made to be contained in washing in elution flow container with elution peristaltic pump coutroi velocity De- liquid enters the intermediate space inside filter core from water sample water outlet with the flow velocity that sets, to make eluent in a manner of backwashing The space between filter core and filter core sleeve is reached, the process of backwash is completed;
3) after process subject to backwash, water sample water inlet and water sample water outlet is closed, will be contained in the step 2) The filtration fraction of eluent using ultrasound or concussion carry out elutriation processing, the eluent after elutriation is handled in filtration fraction from The discharge of water sample water inlet, collects eluent, pathogenic microorganism after being enriched with.
Preferably, in the step 1) water body to be detected include tap water, water supply water, sewage, recycled water, it is secondary go out Water, underground water, surface water, precipitation or seawater.
Preferably, it is 4.5~9.8 that the eluent, which is pH value, the non-ionic surface that mass concentration is 0.01~0.1% The phosphate buffer or Tris salt buffer of activating agent Tween-20.
The present invention provides pathogenic microorganism efficiently concentrating devices in a kind of water environment, including filtration fraction and control unit Point, wherein using peristaltic pump to control water sample to be detected enters sleeve and filter by the water sample water inlet on sleeve with certain flow velocity Space between core, and enter by filter element filtering face the intermediate cylinder of filter core, it is discharged from water sample water outlet.During elutriation, Peristaltic pump controls eluent back through filter core by antiport, to realize washing for the microorganism for being enriched in cartridge surface It is de-.Enriching apparatus provided by the invention has by selecting the filter core of different pore size to realize the enrichment of multiple-microorganism using wide General feature.Enriching apparatus provided by the invention makes full use of the advantages of filter element filtering technology, then complement filter control section and Control section is eluted, enrichment is realized and is eluted in same device and complete, while the set-up mode of filtration fraction of the present invention, meet The filtering of water sample with bulk mass passes to realize the efficiently concentrating of low concentration pathogenic microorganism in water environment for water environment intermediary water Broadcast the detection of disease, monitoring provides technical support.
Enriching apparatus provided by the invention has the characteristics that precision is high, flow is big, structure is uniform simultaneously, further, pottery Porcelain filter core, stainless steel filter core, stud filter core etc. also have the advantages such as corrosion resistance, wearability be strong, and it can be used repeatedly, significantly reduces Use cost.
The present invention provides a kind of method based on pathogenic microorganism in described device enriched water environment, the method operations Simplicity, effect is reliable, it is big to be capable of handling volume of water sample, and can recycle reuse, production cost is effectively reduced.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of enriching apparatus of the present invention, in figure:1. filter core;2. filter core sleeve;3. water sample mistake Peristaltic pump is used in filter;4. water sample water inlet;5. water sample water outlet;6. threeway;7. peristaltic pump is used in elution;8. second one-way valve;9. washing De- flow container;10. cabinet;11. the first check valve.
Specific embodiment
The present invention provides pathogenic microorganism efficiently concentrating devices in a kind of water environment, as shown in Figure 1, including filtration fraction The control section and:The filtration fraction includes filter core and filter core sleeve;The filter core is placed in the filter core barrel bore and described There are spaces between filter core and the internal chamber wall of the filter core sleeve;The top of the filter core sleeve is provided with water sample water inlet;Institute The bottom end for stating filter core is provided with water sample water outlet;The inner wall that the water sample water outlet passes through filter core sleeve extends to outside;It is described Control section includes threeway, water sample filtering control section and elution control section;Water sample filtering control section is according to water sample Elution order has been sequentially communicated the first check valve and water sample filtering peristaltic pump;The opening direction of first check valve is along water Spline filter is opened with peristaltic pump direction;The elution control section according to eluent inflow direction be sequentially communicated elution flow container, Elution peristaltic pump and second one-way valve;The opening direction of the second one-way valve is opened along water sample water outlet direction; The second one-way valve of the water sample water outlet, the first check valve of water sample filtering control section and elution control section passes through threeway Connection.
In the present invention, the material of the filter core sleeve preferably includes stainless steel, glass, PVC or engineering plastics material.
In the present invention, the aperture of the filter core preferably includes 0.45~1 μm.It is selected not according to different types of microorganism Filter core with aperture is enriched with.The filter core preferably includes stainless steel filter core, stud filter core or ceramic element.
In the present invention, all connections are preferably connected to using conduit in the control section.The material of the conduit is preferred Including emulsion tube, silicone tube, PVC hose etc..The present invention is not particularly limited the method for the connection, using known to field Connectivity scheme.
In a preferred embodiment of the invention, the enriching apparatus further includes cabinet;The filtration fraction and control unit quartile In the cabinets cavity.The filtration fraction is removably attached in the inner cavity of cabinet.The side being removably secured Method includes screw, hasp or pipe collar etc..The water sample water inlet extends on the outside of the cabinet.
The present invention provides a kind of methods based on pathogenic microorganism in described device enriched water environment, including following step Suddenly:
1) flow velocity that water sample to be detected is controlled with water sample filtering with peristaltic pump, makes water sample to be detected pass through water with certain flow rate Sample water inlet enters the cavity between filter core sleeve and filter core, then realizes and filters through filtration core, and filtered water sample is from water sample Water outlet discharge;
2) after water sample to be detected filters, made to be contained in washing in elution flow container with elution peristaltic pump coutroi velocity The certain flow velocity of de- liquid enters the intermediate space inside filter core from water sample water outlet, so that de- liquid be made to reach in a manner of backwashing The process of backwash is completed in space between filter core and filter core sleeve;
3) after process subject to backwash, water sample water inlet and water sample water outlet is closed, will be contained in the step 2) The filtration fraction of eluent using ultrasound or concussion carry out elutriation processing, the eluent after elutriation is handled in filtration fraction from The discharge of water sample water inlet, collects eluent, obtains being collected into centrifuge tube containing the eluent of pathogenic microorganism after enrichment, use In the detection of next step.
The present invention controls the flow velocity of water sample to be detected with water sample filtering with peristaltic pump, and water sample to be detected is made to intake by water sample Mouth enters in filter core sleeve, through filter element filtering, is discharged by filtered water sample from water sample water outlet.
In the present invention, the water body to be detected preferably include tap water, water supply water, sewage, recycled water, it is secondary go out Water, underground water, surface water, precipitation or seawater.The flow velocity is preferably not higher than 5L/min, more preferably 2~4L/min, optimal It is selected as 3L/min.
In the present invention, when the process filter element filtering, pathogenic microorganism is retained in filter element filtering in water sample to be detected On face.The selection in the aperture of the filter core is carried out according to the size of pathogenic microorganism to be enriched with, preferred generally for pathogen The filter core in 0.45 μm of aperture, for the filter core in preferably 1.0 μm apertures of protozoon.
In the present invention, the volume of water sample to be measured is handled according to the water quality of pending water:For tap water, water supply It is preferably 50~100L that water, underground water, precipitation etc., which handle volume,;For sewage, recycled water, secondary water-supply, surface water, seawater etc. Handling volume is preferably 10~50L.
After water sample filtering to be detected, present invention elution peristaltic pump coutroi velocity will be contained in elution flow container Eluent enters the space among filter core from water sample water outlet, and the eluent reaches filter core and filter core set in a manner of backwashing In space between cylinder, the filtration fraction containing eluent is obtained.
In the present invention, the flow velocity is preferably not higher than 5L/min, more preferably 2~4L/min, most preferably 3L/ min.The eluent is preferably that pH value is 4.5~9.8, the nonionic surface active agent that mass concentration is 0.01~0.1% Tween-20 phosphate buffer or Tris salt buffer.The time of the elution is preferably 5~30min, more preferably 10min.Eluent will be enriched with when in the space that the eluent reaches between filter core and filter core sleeve in a manner of backwashing After the pathogenic microorganism in filter element filtering face elutes, the eluent containing pathogenic microorganism after enrichment is obtained.
After process subject to backwash, water sample water inlet and water sample water outlet are closed, the present invention described will contain elution The filtration fraction of liquid carries out elutriation processing using ultrasound or concussion, and the eluent after elutriation is handled in filtration fraction is from water sample Water inlet discharge, collects eluent, obtains being collected into centrifuge tube, being used for down containing the eluent of pathogenic microorganism after enrichment The detection of one step.
In the present invention, the parameter of the ultrasonic treatment is as follows:Ultrasonic power is in 30~100KHz, preferably 40KHz, time For 5~45min, temperature is 10~45 DEG C.The present invention is not particularly limited the excusing from death processing with instrument, using this field institute In well known ultrasonic cleaning equipment.
In the present invention, the rate of recovery statistical method of pathogenic microorganism is as follows in the water environment:The eluent that will be obtained It is concentrated, is observed under the microscope or pathogenic microorganism is counted using selective medium, count the micro- life of cause of disease The quantity of object;The quantity of obtained pathogenic microorganism is substituted into formula (1) to the recycling that pathogenic microorganism in water environment is calculated Rate.
The cause of disease added in the rate of recovery (%)=pathogenic microorganism number/water sample to be measured of pathogenic microorganism in water environment The quantity formula (1) of microorganism
Pathogenic microorganism efficiently concentrating device in a kind of water environment provided by the invention is carried out below with reference to embodiment detailed Thin explanation, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
The enrichment of Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst in 100L tap water
It takes 100L city tap-water, after 121 DEG C of high pressure sterilizations, Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst, people is added Work pollutes tap water, its concentration is made to respectively reach 1/L.Select 1.0 μm of aperture filter core, starting water sample filtering peristaltic pump 3, control Water sample flow velocity 2L/min processed makes water sample enter filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, to water sample mistake Stop filtering peristaltic pump 3 after having filtered.
Later, eluent is packed into elution flow container 9, starts elution peristaltic pump 7, eluent is made to pass through filter core intermediate gaps And enriched microorganism is eluted by back flush mode, gap between filter core 1 and filter core sleeve 2 is fully entered to eluent Afterwards, filter core sleeve 2 and filter core 1 are removed, inlet and outlet is closed with clip, places it in ultrasonic cleaning machine or oscillator Elution, recycling eluent carry out Cryptosporidium parvum oocysts suspended and the detection of giardia lamblia stiles sporangiocyst, Cryptosporidium parvum oocysts suspended and giardia lamblia stiles The rate of recovery of sporangiocyst reaches 25%~100%.
Embodiment 2
The enrichment of salmonella and Shigella in 100L tap water
It takes 100L city tap-water, after 121 DEG C of high pressure sterilizations, salmonella and Shigella bacterium solution, artificial contamination is added Tap water makes its concentration respectively reach 10CFU/L.Select 0.45 μm of aperture filter core, starting water sample filtering peristaltic pump 3, control Water sample flow velocity 2L/min makes water sample enter filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, filters to water sample Stop filtering peristaltic pump 3 after complete.
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, eluent is made to pass through filter core 1 Intermediate gaps and the elution that enriched microorganism is carried out by back flush mode, fully enter filter core 1 and filter core sleeve 2 to eluent Between behind gap, remove filter core sleeve 2 and filter core 1, close water sample water inlet 4 and water sample water outlet 5 with clip, place it in super It is eluted in sound washer or oscillator, recycling eluent carries out salmonella and Shigella detection, salmonella and shiga The bacterium rate of recovery respectively reaches 60% or more.
Embodiment 3
The enrichment of Leptospira in 100L tap water
It takes 100L city tap-water, after 121 DEG C of high pressure sterilizations, Leptospira polypide is added, artificial contamination's water sample makes it Concentration reaches 1/L.1.0 μm of aperture filter core is selected, starting water sample filtering peristaltic pump 3 controls water sample flow velocity 2L/min, makes water Sample enters filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, stops filtering peristaltic pump after water sample has filtered 3。
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, pass through eluent in filter core Between gap and the elution of enriched microorganism is carried out by back flush mode, to eluent fully enter filter core 1 and filter core sleeve 2 it Between behind gap, remove filter core sleeve 2 and filter core 1, close water sample water inlet 4 and water sample water outlet 5 with clip, place it in ultrasound It is eluted in washer or oscillator, recycling eluent carries out Leptospira detection, and the rate of recovery reaches 35% or more.
Embodiment 4
The enrichment of Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst in 50L drinking water resource
It takes 50L drinking water resource (underground water), after 121 DEG C of high pressure sterilizations, Cryptosporidium parvum oocysts suspended and giardia lamblia stiles is added Sporangiocyst, artificial contamination's tap water make its concentration respectively reach 1/L.1.0 μm of aperture filter core is selected, starting water sample is filtered with compacted Dynamic pump 3, controls water sample flow velocity 2L/min, so that water sample is entered filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, Stop filtering peristaltic pump 3 after water sample has filtered.
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, eluent is made to pass through filter core 1 Intermediate gaps and the elution that enriched microorganism is carried out by back flush mode, fully enter filter core 1 and sleeve filter core 2 to eluent Between behind gap, remove filter core sleeve 2 and filter core 1, close water sample water inlet 4 and water sample water outlet 5 with clip, place it in super It is eluted in sound washer or oscillator, recycling eluent carries out Cryptosporidium parvum oocysts suspended and the detection of giardia lamblia stiles sporangiocyst, hidden spore The rate of recovery of worm's ovum capsule and giardia lamblia stiles sporangiocyst reaches 25%~100%.
Embodiment 5
The enrichment of salmonella and Shigella in 50L drinking water resource
It takes 50L drinking water resource (underground water), after 121 DEG C of high pressure sterilizations, salmonella and Shigella bacterium solution is added, makes Its concentration respectively reaches 10CFU/L.0.45 μm of aperture filter core is selected, starting water sample filtering peristaltic pump 3 controls water sample flow velocity 2L/min makes water sample enter filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, is stopped after water sample has filtered Filtering peristaltic pump 3.
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, pass through eluent in filter core Between gap and the elution of enriched microorganism is carried out by back flush mode, fully enter filter core 1 to eluent and filter core sleeve 2 filtered Between core behind gap, filter core sleeve 2 and filter core 1 are removed, water sample water inlet 4 and water sample water outlet 5 is closed with clip, places it in It is eluted in ultrasonic cleaning machine or oscillator, recycling eluent carries out salmonella and Shigella detection, and salmonella and will are congratulated The Salmonella rate of recovery reaches 60% or more.
Embodiment 6
The enrichment of Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst in 20L sewage
It takes 20L city domestic sewage, after 121 DEG C of high pressure sterilizations, Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst, people is added Work pollutes tap water, its concentration is made to respectively reach 1/L.Select 1.0 μm of aperture filter core, starting water sample filtering peristaltic pump 3, control Water sample flow velocity 2L/min processed makes water sample enter filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, to water sample mistake Stop filtering peristaltic pump 3 after having filtered.
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, pass through eluent in filter core Between gap and the elution of enriched microorganism is carried out by back flush mode, to eluent fully enter filter core 1 and filter core sleeve 2 it Between behind gap, remove filter core sleeve 2 and filter core 1, close water sample water inlet 4 and water sample water outlet 5 with clip, place it in ultrasound It is eluted in washer or oscillator, recycling eluent carries out Cryptosporidium parvum oocysts suspended and the detection of giardia lamblia stiles sporangiocyst, Cryptosporidium The rate of recovery of egg capsule and giardia lamblia stiles sporangiocyst reaches 15%~100%.
Embodiment 7
The enrichment of salmonella and Shigella in 20L sewage
It takes 20L city domestic sewage, after 121 DEG C of high pressure sterilizations, salmonella and Shigella bacterium solution is added, makes its concentration Respectively reach 10CFU/L.0.45 μm of aperture filter core is selected, starting water sample filtering peristaltic pump 3 controls water sample flow velocity 2L/min, So that water sample is entered filter core sleeve 2 from water sample water inlet 4, be filtered by filter core 1, stops filtering with compacted after water sample has filtered Dynamic pump 3.
Later, a certain amount of eluent is packed into elution flow container 9, starts elution peristaltic pump 7, eluent is made to pass through filter core 1 Intermediate gaps and the elution that enriched microorganism is carried out by back flush mode, fully enter filter core 1 and filter core sleeve 2 to eluent Between behind gap, remove filter core sleeve 2 and filter core 1, close water sample water inlet 4 and water sample water outlet 5 with clip, place it in super It is eluted in sound washer or oscillator, recycling eluent carries out salmonella and Shigella detection, salmonella and shiga The bacterium rate of recovery reaches, and 50% or more.
Embodiment 8
The enrichment of Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst in 50L recycled water
Take 50L recycled water plant effluent, after 121 DEG C of high pressure sterilizations, Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst are artificial dirty Tap water is contaminated, its concentration is made to respectively reach 1/L.1.0 μm of aperture filter core is selected, starting water sample filtering peristaltic pump 3 controls water Sample flow velocity 2L/min makes water sample enter filter core sleeve 2 from water sample water inlet 4, is filtered by filter core 1, has been filtered to water sample Stop filtering peristaltic pump 3 afterwards.
Later, a certain amount of eluent is packed into elution flow container 9, and filter core sleeve 2 and filter core 1 is removed into reversed fixation, opened Dynamic elution peristaltic pump 7, making eluent by 1 intermediate gaps of filter core and by back flush mode progress, enriched microorganism is washed It is de-, it is fully entered between filter core 1 and filter core sleeve 2 behind gap after eluent, removes filter core sleeve 2 and filter core 1, closed with clip Water sample water inlet 4 and water sample water outlet 5, place it in ultrasonic cleaning machine or oscillator and elute, and recycling eluent carries out hidden spore Sub- worm's ovum capsule and the detection of giardia lamblia stiles sporangiocyst, the two rate of recovery reach 15% or more.
Embodiment 9
The enrichment of salmonella and Shigella in 50L recycled water
It takes 50L recycled water plant effluent, after 121 DEG C of high pressure sterilizations, salmonella and Shigella bacterium solution is added, makes its concentration Respectively reach 10CFU/L.0.45 μm of aperture filter core is selected, starting water sample filtering peristaltic pump 3 controls water sample flow velocity 2L/min, So that water sample is entered filter core sleeve 2 from water sample water inlet 4, be filtered by filter core 1, stops filtering with compacted after water sample has filtered Dynamic pump 3.
Later, a certain amount of eluent is packed into elution flow container 9, and filter core sleeve 2 and filter core 1 is removed into reversed fixation, opened Dynamic elution peristaltic pump 7, making eluent by 1 intermediate gaps of filter core and by back flush mode progress, enriched microorganism is washed It is de-, it is fully entered between filter core 1 and filter core sleeve 2 behind gap after eluent, removes filter core sleeve 2 and filter core 1, closed with clip Water sample water inlet 4 and water sample water outlet 5, place it in ultrasonic cleaning machine or oscillator and elute, and recycling eluent carries out sramana Salmonella and Shigella detection, salmonella and the Shigella rate of recovery reach, and 35% or more.
Embodiment 10
Enrichment of the reuse filter core to Cryptosporidium parvum oocysts suspended and giardia lamblia stiles sporangiocyst in 100L tap water
Used 1.0 μm of filter cores are taken, after being impregnated with dilute acid soln, distilled water cleaning, the hidden spore being subsequently used in tap water The enrichment of sub- worm's ovum capsule and giardia lamblia stiles sporangiocyst, and clean repeatedly, using multiple.Cryptosporidium parvum oocysts suspended and giardia lamblia stiles spore The enriching and recovering rate of capsule reaches 10% -100%.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered It is considered as protection scope of the present invention.

Claims (10)

1. pathogenic microorganism efficiently concentrating device in a kind of water environment, which is characterized in that including filtration fraction and control section:
The filtration fraction includes filter core and filter core sleeve;The filter core is placed in the filter core barrel bore and the filter core and institute State between the internal chamber wall of filter core sleeve that there are spaces;The top of the filter core sleeve is provided with water sample water inlet;The filter core Bottom end is provided with water sample water outlet;The inner wall that the water sample water outlet passes through filter core sleeve extends to outside;
The control section includes threeway, water sample filtering control section and elution control section;The water sample filters control section The first check valve and water sample filtering peristaltic pump have been sequentially communicated according to water sample elution order;The opening square of first check valve To be along water sample filtering peristaltic pump direction open;
The elution control section has been sequentially communicated elution flow container, elution peristaltic pump and the second list according to eluent inflow direction To valve;The opening direction of the second one-way valve is opened along water sample water outlet direction;
The second one-way valve of the water sample water outlet, the first check valve of water sample filtering control section and elution control section passes through Threeway connection.
2. enriching apparatus according to claim 1, which is characterized in that the material of the filter core sleeve includes stainless steel, glass Glass, PVC or engineering plastics material.
3. enriching apparatus according to claim 1, which is characterized in that the aperture of the filter core includes 0.45~1 μm;It is described Filter core includes stainless steel filter core, stud filter core or ceramic element.
4. enriching apparatus according to claim 1, which is characterized in that the mode of all connections uses in the control section Conduit connection;The material of the conduit includes emulsion tube, silicone tube or PVC hose.
5. enriching apparatus described according to claim 1~any one of 4, which is characterized in that the enriching apparatus further includes Cabinet;The filtration fraction and control section are located in the cabinets cavity.
6. enriching apparatus according to claim 5, which is characterized in that the filtration fraction is removably attached to cabinet In inner cavity.
7. enriching apparatus according to claim 6, which is characterized in that the water sample water inlet is extended to described by conduit On the outside of cabinet.
8. a kind of method based on pathogenic microorganism in claim 1~7 any one described device enriched water environment, feature It is, includes the following steps:
1) flow velocity of water sample to be detected is controlled with water sample filtering with peristaltic pump, the flow velocity for setting water sample to be detected passes through water sample Water inlet enters the space between filter core sleeve and filter core, the filtering of water sample is then realized through filtration core, by filtered water sample It is discharged by water sample water outlet;
2) after water sample to be detected filters, the eluent being contained in elution flow container is made with elution peristaltic pump coutroi velocity Enter the intermediate space inside filter core from water sample water outlet with the flow velocity of setting, eluent is made to reach filter core in a manner of backwashing The process of backwash is completed in space between filter core sleeve;
3) after process subject to backwash, water sample water inlet and water sample water outlet are closed, elution will be contained in the step 2) The filtration fraction of liquid carries out elutriation processing using ultrasound or concussion, and the eluent after elutriation is handled in filtration fraction is from water sample Water inlet discharge, collects eluent, pathogenic microorganism after being enriched with.
9. according to the method described in claim 8, it is characterized in that, in the step 1) water body to be detected include tap water, from Come source water, sewage, recycled water, secondary water outlet, underground water, surface water, precipitation or seawater.
10. method according to claim 8 or claim 9, which is characterized in that the eluent is that pH value is 4.5~9.8, quality The phosphate buffer or Tris salt buffer for the nonionic surface active agent Tween-20 that concentration is 0.01~0.1%.
CN201810870930.0A 2018-08-02 2018-08-02 Pathogenic microorganism efficiently concentrating device and its enrichment method in a kind of water environment Pending CN108913544A (en)

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CN113324801A (en) * 2021-05-29 2021-08-31 林孔亮 Sampler for detecting content of two insects in water and sampling method thereof
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Application publication date: 20181130