CN104388523A - Method and device of rapidly measuring total number of colonies in water based on concentration counting - Google Patents
Method and device of rapidly measuring total number of colonies in water based on concentration counting Download PDFInfo
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- CN104388523A CN104388523A CN201410719339.7A CN201410719339A CN104388523A CN 104388523 A CN104388523 A CN 104388523A CN 201410719339 A CN201410719339 A CN 201410719339A CN 104388523 A CN104388523 A CN 104388523A
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Abstract
The invention relates to a method and a device of rapidly measuring the total number of colonies in water based on concentration counting. The method comprises the following steps: filtering water to be detected by an ultrafine pore filter screen, and measuring the total number of colonies in a concentrated solution by virtue of an impedance method, wherein the aperture of the ultrafine pore filter screen is smaller than 0.2 micron. The device comprises an on-line device and an off-line device. Compared with the prior art, the method and the device have the advantages of simpleness, rapidness, low cost and the like.
Description
Technical field
The present invention relates to bacterium colony in water to measure, especially relate to a kind of method based on total number of bacterial colony in the Quick Measuring water gaging of concentrated counting and device.
Background technology
The quality and safety of water is always the major issue that people are concerned about.The event that society water quality causes gets more and more.But water quality measurement has its feature and difficult point: even if 1 contaminated water body, the absolute value of bacterium proportion in water body is still very little; 2, in tap water, bacterial number is very small.According to the regulation of GB17324-2003 [5], in every ml water, total number of bacterial colony is less than 20CFU (see table 1).
Table 1 microbiological indicator
To measure the material of minor amount like this, even if meticulous modern test method and equipment all cannot realize direct measurement.Improve bacterium ratio by training method, oppositely deduce measuring and calculating mode, the bacteria content indirectly obtaining measured liquid is principle and the basic skills of current water quality measurement.This causes existing measuring method and technology to exist, and detection speed is slow, complicated operation, cannot the problems such as on-line real-time measuremen, testing cost be expensive.At present, normal detection time needs 24 ~ 48 hours consuming time, belongs to fast detection method, but be all difficult to the needs meeting people's lives, industrial production and government's management of public health lower than the method for 24 hours.
Lower than the supper-fast Methods for Testing Germ Sum of 1 hour, the development of its research and correlation meter is become very urgent.
The microorganism detection method reported at present has a lot, but universally recognized standard method is traditional cultivation counting process in the world, and other new technology is just gradually in development and perfection.
Microorganism detection technical point is the large class of bioelectrochemistry and abiotic electrochemistry two by Hobson in 1996 etc.The former comprises impedance, and conductometric titration, potentiometry, fuel cell technology, circulation/square wave voltammetry, amperometry etc. detect class methods of charge transfer signal between the electrode that caused by microorganism.The latter comprises the traditional methods such as dry weight, live bacterial count and turbidity, also comprise some special techniques, as: microcalorimetry, fluorescence. antibody technique, radiation measurement method, biloluminescence method, piezoelectric membrane method etc., and the method detecting nucleic acid, protein, lipid derivant, carbon/cellular constituent such as phosphoric acid salt, metabolic enzyme.
Bacteria Detection technology is divided into again by Ivnitski in 2000 etc.: cultivate the general method such as counting process, microscopic examination counting process; Method by detecting physical quantities such as piezoquartz, impedance instrument, redox reaction, optical technology, calorimetry, ultrasonic technologies: the method detecting the cellular constituents such as ATP (noclilucence), DNA, protein, lipid derivant (biochemical method), radio isotope (radiometry).
Within 2003, Alocilja and Radke analyzes the market outlook of U.S. domestic pathogenic microbes detect technology.
Within 2004, Deisingh and Thompson is by detecting the biosensor of infective bacterial, is divided into: four classes such as electrochemical method, high frequency biosensor (as piezoquartz, surperficial ultrasonic technique SAW, QCM (Quartz Crystal Microbalance) QCM, surface plasma body resonant vibration SPR etc.), optical biosensor, Electronic Nose.
Within 2005, Noble and Weisberg concludes the fast-bacteria-detection method being applicable to bathing beach water quality analysis, and the feature of these methods can complete detection in 1 hour.According to the object of each several part, whole process generally comprises: captured target bacterium and detection by quantitative two step, in addition for improving detectability, first has a preculture to increase bacterium process toward contact.The seizure of object bacteria can have three class methods: 1. unicellular identification, as with immunoassay, particular molecule probe; 2. nuclei aoid methods, as polymerase chain reaction PCR, quantitative PCR, NASBA: 3. enzyme/substrate approach.Last testing process can utilize the principles such as optics, electrochemistry or piezoelectricity to realize.Except above-mentioned basic skills, some methods with practical prospect utilizing multiple technologies to combine of document selective analysis, as: the method for double wave length fluorescent assay method, immunoassay, PCR-based technology.
Lazcka in 2007 etc. analyze the situation and trend summarizing the application of pathogenic microbes detect technology, according to the height of cognitive ubiquity, are mainly divided into several large classes such as round pcr, cultivation and colony counting method, elisa technique, biosensor, electrophoretic technique.Wherein biosensor has again optics, electrochemistry, piezoelectricity etc. dissimilar by principle of signal conversion.
The Quick Measurement of current total plate count generally uses impedance method.The essence of the method is: the cultivation different bacteriums being carried out to certain hour, to make significant change of properties occurs between substratum and electrode.Reason is: be 10 with impedance method Monitoring lower-cut
3cFU, the tap water bacteria content qualified according to GB is less than 20CFU/mL.When colony-forming unit (CFU, Colony-Forming Units) value reaches impedance method Monitoring lower-cut and afterwards, there is considerable change in detected value.Calculate and detect duration needed for initial time to generation considerable change, according to different colony growth rule calculating formula, can oppositely release total plate count initial value.But when multiple types bacterium exists, the difference of each bacterial classification proportion will affect the precision of impedance method.
Carefully analyze, impedance method has two large defects: 1. need to cultivate.The problems such as this will cause some row problems, as expensive in nutrient solution selection, complicated operation, testing expense, cannot directly to survey online; 2. Measuring Time is long, need calculate with percentage clock.
Summary of the invention
Object of the present invention be exactly in order to overcome above-mentioned prior art exist defect and a kind of method based on total number of bacterial colony in the Quick Measuring water gaging of concentrated counting and device are provided.
Object of the present invention can be achieved through the following technical solutions: a kind of method based on total number of bacterial colony in the Quick Measuring water gaging of concentrated counting, it is characterized in that, the method is that gained concentrated solution measures total plate count wherein by impedance method by water to be detected by ultra-fine hole filter screen filtration; The aperture < 0.2 μm of described ultra-fine hole filtering net.
The aperture > 0.4nm of described ultra-fine hole filtering net.The diameter of water molecules is 0.4nm, and the diameter of general bacterium is between 0.5 ~ 5 μm, and minimum has 0.2 μm, much larger than water molecules.That is most of bacterium can be separated with water by ultra-fine hole filtering net easily, can realize the higher test fluid of bacterial concentration thus, no longer needs to increase bacterium process, convenient direct-detection extremely fast.
A kind of on-line measurement device implementing described method, this device is arranged in drinking water system, it is characterized in that, this device includes water pipe, rising pipe, blow-off pipe, blowdown pipe valve, ultra-fine hole filtering net, test electrode pair, flow valve, described inhalent siphon connects the water source water inlet of drinking water system, described rising pipe connects the water-in of drinking water system, described ultra-fine hole filtering net is arranged between inhalent siphon and rising pipe, and test electrode pair is provided with on the filtering net of ultra-fine hole, described blow-off pipe is communicated to above the filtering net of ultra-fine hole, described blowdown pipe valve is arranged on blow-off pipe, described flow valve is arranged on inhalent siphon.
Described flow valve regulates hydraulic pressure above the filtering net of ultra-fine hole to be 0.1 ~ 0.4MPa.Described inhalent siphon can connect and topping-up pump, and above making ultra-fine hole filtering net, hydraulic pressure ensures at 0.1 ~ 0.4MPa.
Described inhalent siphon, rising pipe, blow-off pipe, blowdown pipe valve, flow valve all adopt the fungi-proofing material of health to make, and the fungi-proofing material of described health comprises clear quartz material.
Described test electrode is to connection impedance method test set.Impedance method test set is existing commercial device.
A kind of non-on-line measurement device implementing described method, it is characterized in that, this device comprises quartz cavity cylinder with a scale, inhalent siphon, tongs, pressure piston, ultra-fine hole filtering net, potential electrode pair, described inhalent siphon is arranged on quartz cavity cylinder sidewall, and tongs is provided with on inhalent siphon, described pressure piston inserts from quartz cavity cylinder top, and described ultra-fine hole filtering net is arranged in quartz cavity cylinder, and described potential electrode is to being arranged on the filtering net of ultra-fine hole.Adopt pressure piston to apply pressure, above making ultra-fine hole filtering net, hydraulic pressure ensures at 0.1 ~ 0.4MPa.
The size and shape of described ultra-fine hole filtering net is identical with the cross section of described quartz cavity cylinder.Quartz cavity cylinder and pressure piston are that prototype realizes with reference to glass syringe, and during manufacture, tongs, ultra-fine hole filtering net are HEPA micro-filtration net and all will possess the ability of bearing 0.1 ~ 0.4MPa pressure difference.
Compared with prior art, the present invention is directed to the feature of tap water: 1, the scale of construction is large, 2, water molecules diameter is 0.4 nanometer, bacterium bore is more than 500 nanometers, much larger than water molecules diameter, 3, need testing liquid to cultivate the long period during impedance measurement in nutrient solution, make bacterial concentration reach certain C FU value can detect etc., on market study basis, propose based on " concentrated counting " but not cultivate the novel method counted, namely based on micro-filtration net pack bacterium, the supper-fast total number of bacterial colony electric measuring method of incubation time is exempted.Under novel method instructs, devise a online measuring apparatus, the cheap rapid measurement device of a manual type, the simple and quick total number of bacterial colony measured at low cost in water.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of the online total number of bacterial colony measuring apparatus of the present invention;
Fig. 2 is the schematic diagram of the non-online total number of bacterial colony measuring apparatus of the present invention.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in detail.
Embodiment 1
As shown in Figure 1, a kind of on-line measurement device implemented based on total number of bacterial colony method in the Quick Measuring water gaging of concentrated counting, this device is arranged in drinking water system, this device includes water pipe 1, rising pipe 2, blow-off pipe 3, blowdown pipe valve 4, ultra-fine hole filtering net 5, test electrode is to 6, flow valve, described inhalent siphon 1 connects the water source water inlet 7 of drinking water system, described rising pipe 2 connects the water-in 8 of drinking water system, described ultra-fine hole filtering net 5 is arranged between inhalent siphon 1 and rising pipe 2, and test electrode is provided with to 6 on ultra-fine hole filtering net 5, described blow-off pipe 3 is communicated to above the filtering net of ultra-fine hole, described blowdown pipe valve 4 is arranged on blow-off pipe 3, described flow valve is arranged on inhalent siphon, it is 0.1 ~ 0.4MPa that adjust flux valve controls hydraulic pressure above ultra-fine hole filtering net 5.Described inhalent siphon can connect and topping-up pump, and above making ultra-fine hole filtering net, hydraulic pressure ensures at 0.1 ~ 0.4MPa.
Described inhalent siphon, rising pipe, blow-off pipe, blowdown pipe valve, flow valve all adopt the fungi-proofing material of health to make, and the fungi-proofing material of described health comprises clear quartz material.
The aperture < 0.2 μm of 0.4nm < ultra-fine hole filtering net.The diameter of water molecules is 0.4nm, and the diameter of general bacterium is between 0.5 ~ 5 μm, and minimum has 0.2 μm, much larger than water molecules.That is most of bacterium can be separated with water by ultra-fine hole filtering net easily, can realize the higher test fluid of bacterial concentration thus, no longer needs to increase bacterium process, convenient direct-detection extremely fast.The sealing of bend pipe and electrode leads to client, sprays with waterproof.
During test, water to be detected is filtered by ultra-fine hole filtering net 5, be 0.1 ~ 0.4MPa by the topping-up pump hydraulic pressure controlled above ultra-fine hole filtering net 5, above ultra-fine hole filtering net 5, obtain concentrated solution, this concentrated solution is by connecting the test electrode of existing impedance method measuring apparatus to 6 measurements total plate count wherein; Calculate according to the circulation of water to be detected and the amount of concentrated solution, the amount of bacteria in water to be detected.
Embodiment 2
As shown in Figure 2, a kind of non-on-line measurement device implemented based on total number of bacterial colony method in the Quick Measuring water gaging of concentrated counting, this device comprises quartz cavity cylinder 21 with a scale, inhalent siphon 22, tongs 23, pressure piston 24, ultra-fine hole filtering net 25, potential electrode is to 26, described inhalent siphon 22 is arranged on quartz cavity cylinder 21 sidewall, and tongs 23 is provided with on inhalent siphon 22, described pressure piston 24 inserts from quartz cavity cylinder 21 top, described ultra-fine hole filtering net 25 is arranged in quartz cavity cylinder 21, the size and shape of described ultra-fine hole filtering net 25 is identical with the cross section of described quartz cavity cylinder 21.Described potential electrode is arranged on ultra-fine hole filtering net 25 26.Quartz cavity cylinder 21 and pressure piston 24 are that prototype realizes with reference to glass syringe, and during manufacture, tongs, ultra-fine hole filtering net are HEPA micro-filtration net and all will possess the ability of bearing 0.1 ~ 0.4MPa pressure difference.The aperture < 0.2 μm of 0.4nm < ultra-fine hole filtering net.
During use, pressure is applied by pressure piston 24, above making ultra-fine hole filtering net 25, hydraulic pressure ensures at 0.1 ~ 0.4MPa, above ultra-fine hole filtering net 25, obtain concentrated solution, and this concentrated solution is by connecting the test electrode of existing impedance method measuring apparatus to 26 measurements total plate count wherein; Calculate according to the total amount of water to be detected and the amount of concentrated solution, the amount of bacteria in water to be detected.
As the test fluid of 100mL, by filtering, obtain 1ml concentrated filtrate, if recording its total plate count is 2000CFU/mL, then the total plate count that can calculate test fluid is easily 20CFU/mL.
Claims (8)
1. based on a method for total number of bacterial colony in the Quick Measuring water gaging of concentrated counting, it is characterized in that, the method is that gained concentrated solution measures total plate count wherein by impedance method by water to be detected by ultra-fine hole filter screen filtration; The aperture < 0.2 μm of described ultra-fine hole filtering net.
2. a kind of method based on total number of bacterial colony in the Quick Measuring water gaging of concentrated counting according to claim 1, is characterized in that, the aperture > 0.4nm of described ultra-fine hole filtering net.
3. one kind implements the claims the on-line measurement device of method described in 1, this device is arranged in drinking water system, it is characterized in that, this device includes water pipe, rising pipe, blow-off pipe, blowdown pipe valve, ultra-fine hole filtering net, test electrode pair, flow valve, described inhalent siphon connects the water source water inlet of drinking water system, described rising pipe connects the water-in of drinking water system, described ultra-fine hole filtering net is arranged between inhalent siphon and rising pipe, and test electrode pair is provided with on the filtering net of ultra-fine hole, described blow-off pipe is communicated to above the filtering net of ultra-fine hole, described blowdown pipe valve is arranged on blow-off pipe, described flow valve is arranged on inhalent siphon.
4. on-line measurement device according to claim 3, is characterized in that, described flow valve regulates hydraulic pressure above the filtering net of ultra-fine hole to be 0.1 ~ 0.4MPa.
5. on-line measurement device according to claim 3, is characterized in that, described inhalent siphon, rising pipe, blow-off pipe, blowdown pipe valve, flow valve all adopt the fungi-proofing material of health to make, and the fungi-proofing material of described health comprises clear quartz material.
6. on-line measurement device according to claim 3, is characterized in that, described test electrode is to connection impedance method test set.
7. one kind implements the claims the non-on-line measurement device of method described in 1, it is characterized in that, this device comprises quartz cavity cylinder with a scale, inhalent siphon, tongs, pressure piston, ultra-fine hole filtering net, potential electrode pair, described inhalent siphon is arranged on quartz cavity cylinder sidewall, and tongs is provided with on inhalent siphon, described pressure piston inserts from quartz cavity cylinder top, and described ultra-fine hole filtering net is arranged in quartz cavity cylinder, and described potential electrode is to being arranged on the filtering net of ultra-fine hole.
8. non-on-line measurement device according to claim 7, is characterized in that, the size and shape of described ultra-fine hole filtering net is identical with the cross section of described quartz cavity cylinder.
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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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| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
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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 |
| 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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Patent Citations (4)
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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 |
| 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 |
Non-Patent Citations (1)
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| 张甲耀等: "《环境微生物学下册》", 31 December 2008 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109439536A (en) * | 2017-08-18 | 2019-03-08 | 黄国仁 | Drink water bacteria fast culture device and total plate count rapid detection system |
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