CN104388523B - The method and device of total plate count in a kind of quick measurement water counted based on concentration - Google Patents
The method and device of total plate count in a kind of quick measurement water counted based on concentration Download PDFInfo
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- CN104388523B CN104388523B CN201410719339.7A CN201410719339A CN104388523B CN 104388523 B CN104388523 B CN 104388523B CN 201410719339 A CN201410719339 A CN 201410719339A CN 104388523 B CN104388523 B CN 104388523B
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Abstract
The present invention relates to a kind of method and device of total plate count in quick measurement water counted based on concentration, the method is that, by ultra-fine hole filter screen filtration, gained concentrate measures total number of bacteria therein by impedance method by water to be detected；0.2 μm of the aperture ＜ of described ultra-fine hole screen pack.The device of use includes on-line equipment and non-on-line equipment.Compared with prior art, the present invention has simple, quick, inexpensive low advantage.
Description
Technical field
Measured the present invention relates to bacterium colony in water, it is total more particularly, to bacterium colony in a kind of quick measurement water counted based on concentration
Several method and devices.
Background technology
The quality and safety of water is always the major issue that people are concerned about.The event that today's society water quality causes is increasingly
It is many.But water quality measurement has its feature and difficult point：1st, even contaminated water body, the absolute value of bacterium proportion in water body
Still very little；2nd, in drinking water, bacterial number is very small.According to the regulation of GB17324-2003 [5], bacterium colony is total in every milliliter of water
Number is less than 20CFU (being shown in Table 1).
The microbiological indicator of table 1
The material of such minor amount is measured, even fine modern test method and equipment cannot be real
Existing direct measurement.Bacterium ratio is improved by training method, measuring and calculating mode is reversely deduced, indirect gain fluid to be measured it is thin
Bacterial content is the principle and basic skills of current water quality measurement.This cause existing measuring method and technology exist detection speed it is slow,
Complex operation, cannot on-line real-time measuremen, the problems such as testing cost is expensive.At present, need to take 24~48 small for normal detection time
When, the method less than 24 hours belongs to fast detection method, but is difficult to meet people's lives, industrial production and government's public health
The need for management.
Supper-fast Methods for Testing Germ Sum less than 1 hour, studies it and the development of correlation meter becomes
It is very urgent.
The microorganism detection method reported at present has a lot, but universally recognized standard method is traditional training in the world
Counting method is supported, other new technologies are just gradually in development and perfection.
Microorganism detection technology is divided into bioelectrochemistry and the major class of abiotic electrochemistry two by Hobson in 1996 etc..The former
Including impedance, conductance method, potentiometry, fuel cell technology, circulation/square wave voltammetry, amperometry etc. are detected by micro-
One class method of charge transfer signal between the electrode that biology causes.The latter includes dry weight, count plate and conventional method etc. turbidity,
Also include some special techniques, such as：Microcalorimetric method, fluorescence antibody techniques, actinometry method, biloluminescence method, piezoelectric membrane method
Etc., and the method for detecting the cell components such as nucleic acid, protein, lipid derivant, carbon/phosphate, metabolic enzyme.
Be divided into for Bacteria Detection technology again by Ivnitski in 2000 etc.：Culture counting method, micro- sem observation counting method etc. one
As method；Physical quantity is detected with piezo-electric crystal, impedance instrument, redox reaction, optical technology, calorimetry, ultrasonic technology etc.
Method：Detection ATP (bioluminescence), DNA, protein, lipid derivant (biochemical method), radio isotope (spoke
Penetrate tolerance) etc. cell component method.
Alocilja and Radke analyzes the market prospects of U.S. domestic pathogenic bacteria detection technique within 2003.
Deisingh and Thompson in 2004 will detect the biology sensor of infective bacterial, be divided into：Electrochemistry side
(such as piezo-electric crystal, surface ultrasonic technique SAW, QCM QCM, surface plasma are common for method, high frequency biology sensor
Shake SPR etc.), optical biosensor, four classes such as Electronic Nose.
Noble and Weisberg in 2005 is carried out to the fast-bacteria-detection method suitable for bathing beach water analysis
The characteristics of conclusion, these methods is that detection can be completed in 1 hour.According to the purpose of each several part, whole process is generally comprised：Catch
Object bacteria and the step of quantitative determination two are caught, in addition to improve test limit, first there is a preculture to increase bacterium process toward contact.Object bacteria
Seizure can have three class methods：1. unicellular identification, such as with immunoassay, particular molecule probe；2. nuclei aoid methods, are such as polymerized
PCR PCR, quantitative PCR, NASBA：3. enzyme/substrate approach.Last detection process can utilize optics, electrochemistry or pressure
The principles such as electricity are realized.In addition to above-mentioned basic skills, document selective analysis some using multiple technologies combine with practicality before
The method of scape, such as：Double wave length fluorescent determination method, immunoassay, the method for PCR-based technology.
The analysis such as Lazcka in 2007 summarizes the present situation and trend of pathogenic bacteria detection technique application, according to cognitive generality
Height, be mainly divided into round pcr, culture several with colony counting method, elisa technique, biology sensor, electrophoretic techniques etc. big
Class.Wherein biology sensor is had the different types such as optics, electrochemistry, piezoelectricity by principle of signal conversion again.
Impedance method is generally used in the quick measurement of current total number of bacteria.The essence of the method is：Different bacteriums is entered
The culture of row certain hour, so as to there is significant change of properties between culture medium and electrode.Reason is：Under being detected with impedance method
It is limited to 103CFU, 20CFU/mL is less than according to the qualified drinking water bacterial content of GB.When CFU (CFU,
Colony-Forming Units) value reaches impedance method Monitoring lower-cut and afterwards, and there is significant change in detected value.Calculating is detected
Begin duration needed for the moment to generation significant change, according to different colony growth rule calculating formulas, at the beginning of reversely can releasing total number of bacteria
Initial value.But, in the presence of multiple types bacterium, the different of each strain proportion will influence the precision of impedance method.
Carefully analyze, impedance method has two big defects：1. culture is needed.This will cause some row problems, such as nutrient solution choosing
Select, the problems such as complex operation, testing expense are expensive, cannot directly survey online；2. time of measuring is long, need to be calculated with percentage clock.
The content of the invention
The purpose of the present invention is exactly to provide a kind of based on concentration counting for the defect for overcoming above-mentioned prior art to exist
Quick measurement water in total plate count method and device.
The purpose of the present invention can be achieved through the following technical solutions：In a kind of quick measurement water counted based on concentration
The method of total plate count, it is characterised in that the method is that, by ultra-fine hole filter screen filtration, gained concentrate leads to by water to be detected
Cross impedance method and measure total number of bacteria therein；0.2 μm of the aperture ＜ of described ultra-fine hole screen pack.
The aperture ＞ 0.4nm of described ultra-fine hole screen pack.A diameter of 0.4nm of hydrone, the diameter of general bacterium is about
Between 0.5~5 μm, minimum has 0.2 μm, much larger than hydrone.That is ultra-fine hole screen pack can be easily
Most of bacterium is separated from water, bacterial concentration test fluid higher thus can be realized, it is no longer necessary to increase bacterium process, be conveniently exceedingly fast
The direct detection of speed.
A kind of on-line measurement device for implementing methods described, the device is arranged in drinking water system, it is characterised in that should
Device includes oral siphon, outlet pipe, blow-off pipe, blowdown pipe valve, ultra-fine hole screen pack, test electrode pair, flow valve, and described enters
Water pipe connects the water source water inlet of drinking water system, and described outlet pipe connects the water inlet of drinking water system, and described is ultra-fine
Hole screen pack is arranged between oral siphon and outlet pipe, and test electrode pair, described blowdown are provided with ultra-fine hole screen pack
Pipe is communicated to ultra-fine hole screen pack top, and described blowdown pipe valve is arranged on blow-off pipe, and described flow valve is arranged on into water
Guan Shang.
The ultra-fine hole screen pack top hydraulic pressure of described flow valve regulation is 0.1~0.4MPa.Described oral siphon can be connected
With booster pump so that ultra-fine hole screen pack top hydraulic pressure ensures in 0.1~0.4MPa.
Described oral siphon, outlet pipe, blow-off pipe, blowdown pipe valve, flow valve is made of the fungi-proofing material of health, described
The fungi-proofing material of health include clear quartz material.
Described test electrode pair connection impedance method test device.Impedance method test device is existing commercial device.
A kind of non-on-line measurement device for implementing methods described, it is characterised in that the device includes quartz cavity with a scale
Cylinder, oral siphon, tongs, pressure piston, ultra-fine hole screen pack, measuring electrode pair, described oral siphon are arranged on quartz cavity cylinder
On the wall of side, and tongs is provided with oral siphon, described pressure piston is inserted from quartz cavity cylinder top, described ultra-fine hole mistake
Filter screen is arranged in quartz cavity cylinder, and described measuring electrode is to being arranged on ultra-fine hole screen pack.Apply to press using pressure piston
Power so that ultra-fine hole screen pack top hydraulic pressure ensures in 0.1~0.4MPa.
The size and shape of described ultra-fine hole screen pack is identical with the cross section of quartz cavity cylinder.Quartz cavity cylinder and pressure
Power piston is with reference to glass syringe for prototype realizes that during manufacture, tongs, ultra-fine hole screen pack are HEPA micro-filtration nets and are intended to
Possesses the ability for bearing 0.1~0.4MPa pressure differentials.
Compared with prior art, the characteristics of present invention is for drinking water：1st, the scale of construction is big, and 2, a diameter of 0.4 nanometer of hydrone,
Bacterium bore more than 500 nanometers, much larger than hydrone diameter, 3, impedance measurement when need to cultivate testing liquid in nutrient solution
Long period, bacterial concentration is reached certain C FU values can detect etc., on the basis of market survey, it is proposed that based on " concentration
Count " rather than cultivate the new method for counting, i.e., based on micro-filtration net pack bacterium, exempt the supper-fast total plate count electricity of incubation time
Survey method.Under new method guidance, a online measurement apparatus, the cheap rapid measurement device of a hand are devised, letter
The total plate count in water has been measured single fast and low-cost.
Brief description of the drawings
Fig. 1 is the schematic diagram of online total plate count measurement apparatus of the invention；
Fig. 2 is the schematic diagram of the non-online total plate count measurement apparatus of the present invention.
Specific embodiment
The present invention is described in detail with specific embodiment below in conjunction with the accompanying drawings.
Embodiment 1
As shown in figure 1, the on-line measurement of total plate count method is filled in a kind of quick measurement water for implementing to be counted based on concentration
Put, the device is arranged in drinking water system, the device includes oral siphon 1, outlet pipe 2, blow-off pipe 3, blowdown pipe valve 4, ultra-fine
Hole screen pack 5, test electrode pair 6, flow valve, described oral siphon 1 connect the water source water inlet 7 of drinking water system, described
Outlet pipe 2 connects the water inlet 8 of drinking water system, and described ultra-fine hole screen pack 5 is arranged between oral siphon 1 and outlet pipe 2,
And test electrode pair 6 is provided with ultra-fine hole screen pack 5, described blow-off pipe 3 is communicated to ultra-fine hole screen pack top, described
Blowdown pipe valve 4 is arranged on blow-off pipe 3, and described flow valve is arranged on oral siphon, and regulation flow valve controls ultra-fine hole filtering
The top hydraulic pressure of net 5 is 0.1~0.4MPa.Described oral siphon can be connected and booster pump so that ultra-fine hole screen pack top hydraulic pressure
Ensure in 0.1~0.4MPa.
Described oral siphon, outlet pipe, blow-off pipe, blowdown pipe valve, flow valve is made of the fungi-proofing material of health, described
The fungi-proofing material of health include clear quartz material.
0.2 μm of the aperture ＜ of the ultra-fine hole screen packs of 0.4nm ＜.A diameter of 0.4nm of hydrone, the diameter of general bacterium is about
Between 0.5~5 μm, minimum has 0.2 μm, much larger than hydrone.That is ultra-fine hole screen pack can be easily
Most of bacterium is separated from water, bacterial concentration test fluid higher thus can be realized, it is no longer necessary to increase bacterium process, be conveniently exceedingly fast
The direct detection of speed.The sealing of bend pipe and electrode leads to client, is sprayed with waterproof.
During test, water to be detected is filtered by ultra-fine hole screen pack 5, and the top of ultra-fine hole screen pack 5 is controlled by booster pump
Hydraulic pressure be 0.1~0.4MPa, obtain concentrate in the top of ultra-fine hole screen pack 5, the concentrate is by connecting existing impedance method
The test electrode pair 6 of measurement apparatus measures total number of bacteria therein；The amount of circulation and concentrate according to water to be detected is calculated
Draw, the amount of bacteria in water to be detected.
Embodiment 2
As shown in Fig. 2 a kind of non-on-line measurement for implementing total plate count method in the quick measurement water based on concentration counting
Device, the device includes quartz cavity cylinder 21, oral siphon 22, tongs 23, pressure piston 24, ultra-fine hole screen pack with a scale
25th, to 26, described oral siphon 22 is arranged on quartz cavity 21 side walls of cylinder measuring electrode, and is provided with tongs on oral siphon 22
23, from the top insertion of quartz cavity cylinder 21, described ultra-fine hole screen pack 25 is arranged on quartz cavity cylinder 21 to described pressure piston 24
Interior, the size and shape of described ultra-fine hole screen pack 25 is identical with the cross section of quartz cavity cylinder 21.Described measurement electricity
Extremely it is arranged on to 26 on ultra-fine hole screen pack 25.Quartz cavity cylinder 21 and pressure piston 24 are prototype reality with reference to glass syringe
Existing, during manufacture, tongs, ultra-fine hole screen pack are HEPA micro-filtration nets and are intended to possess the energy for bearing 0.1~0.4MPa pressure differentials
Power.0.2 μm of the aperture ＜ of the ultra-fine hole screen packs of 0.4nm ＜.
When using, pressure being applied by pressure piston 24 so that the top hydraulic pressure of ultra-fine hole screen pack 25 ensures 0.1~
0.4MPa, concentrate is obtained in the top of ultra-fine hole screen pack 25, and the concentrate is by connecting the survey of existing impedance method measurement apparatus
Examination electrode pair 26 measures total number of bacteria therein；The amount of total amount and concentrate according to water to be detected is calculated, water to be detected
In amount of bacteria.
Such as the test fluid of 100mL, by filtering, 1ml concentration filtrates are obtained, if measuring its total number of bacteria for 2000CFU/
ML, the then total number of bacteria that can easily calculate test fluid is 20CFU/mL.
Claims (2)
1. it is a kind of based on concentration count quick measurement water in total plate count on-line measurement device, its measuring method is will be to be checked
Water is surveyed by ultra-fine hole filter screen filtration, gained concentrate measures total number of bacteria therein by impedance method；Described ultra-fine hole
The aperture of screen pack<0.2 μm, described device is arranged in drinking water system, it is characterised in that the device include oral siphon,
Outlet pipe, blow-off pipe, blowdown pipe valve, ultra-fine hole screen pack, test electrode pair, flow valve, described oral siphon connection direct drinking
The water source water inlet of system, described outlet pipe connects the water inlet of drinking water system, and described ultra-fine hole screen pack is arranged on
Between oral siphon and outlet pipe, and test electrode pair is provided with ultra-fine hole screen pack, described blow-off pipe is communicated to ultra-fine hole
Screen pack top, described blowdown pipe valve is arranged on blow-off pipe, and described flow valve is arranged on oral siphon；
The ultra-fine hole screen pack top hydraulic pressure of described flow valve regulation is 0.1~0.4MPa；
Described oral siphon, outlet pipe, blow-off pipe, blowdown pipe valve, flow valve is made of the fungi-proofing material of health, and described defends
Biocontrol microorganisms material includes clear quartz material；
Described test electrode pair connection impedance method test device.
2. it is a kind of based on concentration count quick measurement water in total plate count non-on-line measurement device, its measuring method is to treat
By ultra-fine hole filter screen filtration, gained concentrate measures total number of bacteria therein to detection water by impedance method；Described is ultra-fine
The aperture of hole screen pack<0.2 μm, it is characterised in that described device include quartz cavity cylinder with a scale, oral siphon, tongs,
Pressure piston, ultra-fine hole screen pack, measuring electrode pair, described oral siphon are arranged on the wall of quartz cavity cylinder side, and in oral siphon
Tongs is provided with, from the top insertion of quartz cavity cylinder, described ultra-fine hole screen pack is arranged on quartz cavity to described pressure piston
In cylinder, described measuring electrode is to being arranged on ultra-fine hole screen pack；
The size and shape of described ultra-fine hole screen pack is identical with the cross section of quartz cavity cylinder.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201410719339.7A CN104388523B (en) | 2014-12-02 | 2014-12-02 | The method and device of total plate count in a kind of quick measurement water counted based on concentration |
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| CN109439536A (en) * | 2017-08-18 | 2019-03-08 | 黄国仁 | Drink water bacteria fast culture device and total plate count rapid detection system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101981447A (en) * | 2008-02-14 | 2011-02-23 | 3M创新有限公司 | Methods and compositions for detecting microorganisms |
| CN102954985A (en) * | 2011-08-31 | 2013-03-06 | 中国科学院上海微系统与信息技术研究所 | Data analysis method for electrical impedance type quick bacteria detection sensor |
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| JP2002153297A (en) * | 2000-11-24 | 2002-05-28 | Matsushita Electric Ind Co Ltd | Method for counting microorganisms and microorganisms counter with pretreatment device |
| JP2005210937A (en) * | 2004-01-28 | 2005-08-11 | Mitsubishi Electric Corp | Microorganism number-measuring method and microorganism number-measuring device used therefor |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101981447A (en) * | 2008-02-14 | 2011-02-23 | 3M创新有限公司 | Methods and compositions for detecting microorganisms |
| CN102954985A (en) * | 2011-08-31 | 2013-03-06 | 中国科学院上海微系统与信息技术研究所 | Data analysis method for electrical impedance type quick bacteria detection sensor |
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