JPH05273217A - Measuring apparatus for microbe - Google Patents

Abstract

PURPOSE:To easily collect microbes in a short time and to enhance and stabilize the collecting rate of microbes by controlling a sample feeding means, a transfer device, etc., by a controlling device and automatizing the measurement of microbes. CONSTITUTION:A transportation pipe 14 of a sample liquid connecting a sample liquid tank 1 and a solenoid valve 6 is provided with an electromagnetic valve 2, a flowmeter 3, a pump 4 as a sample liquid feeding means and a pressure gauge 5. When the pump 4 is driven, the sample liquid is fed to a filter cartridge 9 through the solenoid valve 6 and an injection pipe 16. At this time, it is possible to measure the flow rate of the sample liquid by the flowmeter 3 and also the liquid pressure of the sample liquid by the pressure gauge 5. A broken liquid transportation pipe 15 connecting a tank 8 of a liquid of broken microbes with the solenoid valve 6 is provided with a pump 7 which is a distributing means of the liquid of broken microbes. The broken liquid is, through the solenoid valve 6 and the injection pipe 16, injected into the filter cartridge 9 by this pump 7.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for immunologically measuring microorganisms, and more particularly to pretreatment for measuring the number of microorganisms in sample water.

[0002]

2. Description of the Related Art Conventionally, the measurement of microorganisms in sample water has been carried out by concentrating the sample water with a membrane filter and ultrasonically crushing it, and then performing immunoassay.

When the sample water is concentrated with this membrane filter, it is necessary to use a pre-filter in order to shorten the concentration time and remove impurities.

FIG. 3 shows an explanatory view of a microorganism concentrating device using the above prefilter and membrane filter.

As shown in this figure, this microorganism concentrator comprises a funnel 21 for containing sample water, a filter section 22 for filtering and concentrating the sample water, a side flask 23 for containing the filtered and concentrated sample water, and a funnel. It is composed of fixed scissors 26 for fixing 21 and the flask with side branch 23.

FIG. 4 is an enlarged view of the filtration section of this microorganism concentrating apparatus. The filtration section 22 is formed by arranging a funnel 21, an O-ring 24, a membrane filter 25, and a side flask 23 in this order.

In order to concentrate microorganisms with the above apparatus, sample water is put into the funnel 21 and the sample water is sucked into the inside of the side-arm flask by lowering the internal pressure with a suction pump through the side tube of the side-arm flask 27. To do.

At this time, since the microorganisms in the sample water are captured by the membrane filter 5 of the filtration section, the membrane filter 5 is collected, and the buffer solution and the glass beads are added and shaken in the glass beaker.

Thus, the bacteria trapped on the surface of the membrane filter are uniformly suspended in the buffer solution, and the suspension is pipetted and transferred to a sample tube.

Furthermore, the microorganisms in the sample water are measured by ultrasonically disrupting the suspension in the sample tube with an ultrasonic disruptor and then performing enzyme immunoassay.

[0011]

However, in the above-mentioned conventional measuring method, the collecting operation, which is a pretreatment of the enzyme immunoassay, is complicated and the collecting rate is difficult to be constant, and the measuring accuracy becomes low. Further, the recovery operation requires about 15 minutes, which causes a large loss of time.

Further, a large-scale apparatus or instrument such as a filtering apparatus such as a side-branched flask or a funnel and a shaker used at the time of recovery is required, and automation of pretreatment is also difficult.

The present invention has been made under the above-mentioned background, and a microbial immunology capable of performing a microbial recovery operation easily and in a short time and having a high microbial recovery rate and stabilization. An object of the present invention is to provide a method for dynamic measurement and a microorganism filtration container.

[0014]

In order to solve the above problems, the present invention provides a sample liquid tank for storing a sample liquid, a microbial disruption liquid tank for storing a microbial disruption liquid, a sample liquid inlet and a filtered water outlet. A filter cartridge having a microbial capture filter for partitioning the inside of the outer cylinder into a sample water inlet side and a sample liquid outlet side, and a filter cartridge supply device are provided inside the outer cylinder provided, and the filter cartridge is supplied by a transfer device. The filter cartridge obtained from the device is installed at a position where the supply of the sample solution and the microbial disruption solution can be provided, and then the sample solution obtained from the sample solution tank is introduced into the filter cartridge by the sample solution inflow means. After filtering the liquid,
The microbial crushed liquid obtained from the microbial crushed liquid tank is injected by the microbial crushed liquid injecting means, and the filter cartridge in which the microbial crushed liquid is injected is installed in the microbial crusher by the transfer device to perform microbial crushing, A microbe measuring device in which a filter cartridge is installed in a sample liquid sampling device by a transfer device, and the sample liquid that has been subjected to the microbe crushing is sampled by the sample sampling device and then the measuring device measures the microorganisms in the sample liquid. The microorganism measuring device is a control means for controlling the sample liquid inflow means, the microorganism crushing liquid injecting means, the filter cartridge supply device, the transfer device, the microorganism crusher, the sample liquid collecting device, and the measuring device. And characterized by automating the measurement of microorganisms.

Further, in the above-mentioned microorganism measuring apparatus, the sample liquid inflow means has a liquid pressure detecting means for the sample liquid, and the control means flows in the sample liquid based on the liquid pressure obtained by the liquid pressure detecting means. There is also provided a microorganism measuring device characterized by controlling the amount.

Further, in each of the above-mentioned microorganism measuring devices, the microorganism measuring device has means for detecting the inflow amount of the sample liquid and the injecting amount of the microbial disruption liquid in the filter cartridge, and the control device is the inflow amount of the sample liquid. And calculating the concentration rate of the microorganisms based on the inflow rate of the sample solution and the injection rate of the microbial disruption solution obtained from the means for detecting the injection rate of the microbial disruption solution, and to the output unit provided in advance in the microorganism measuring device. There is also provided a microorganism measuring device characterized by outputting a concentration factor.

Furthermore, in each of the above-mentioned microorganism measuring devices, the microorganism measuring device has a sealing means for sealing the sample liquid outflow portion of the filter cartridge, and the sealing means is controlled by the control device.
After the filtration of the sample solution is completed, the sample solution outflow portion of the filter cartridge is sealed by the sealing means, and further, an ultrasonic crusher is used as the microbial crusher, and the filter cartridge is immersed in the ultrasonic crusher. There is also provided a microorganism measuring device characterized by crushing microorganisms by ultrasonically crushing the microorganisms.

[0018]

In the microorganism measuring apparatus according to the present invention, the control apparatus controls the sample inflow means, the transfer apparatus and the like to automate the measurement of microorganisms. The details are shown below.

First, the sample liquid inflow means is operated to filter the sample liquid, and after the filtration is completed, the sample liquid inflow means is stopped. After that, the microorganism crushed liquid injection means is operated to inject the microbial crushed liquid into the filter cartridge.

Next, the transfer device is operated to install the filter cartridge into which the above-mentioned microorganism crushing liquid has been injected into the microbial crusher. After that, the microorganism crusher is operated to crush the microorganisms.

Further, the transfer device is operated to install the filter cartridge in the sample liquid collecting device, and the sample liquid collecting device is operated to collect the sample liquid, and the sample liquid is transported to the measuring device. The microorganisms are measured by operating the measuring device. In the above control device, a device for detecting the liquid pressure of the sample liquid when the sample liquid flows into the filter cartridge is provided, and the sample liquid inflow means is controlled by the value of this liquid pressure to control the inflow amount of the sample liquid. Preferably.

Similarly, it is preferable to detect the inflow amount of the sample liquid and the injection amount of the crushed microorganism liquid in the filter cartridge, and output the concentration ratio of the microorganism obtained from these values.

Further, the microorganism measuring device is provided with a sealing means for the sample liquid outflow portion in the filter cartridge, the sample liquid outflow portion is sealed after filtering the sample liquid, and an ultrasonic disintegrator is used as a microorganism disintegrator. It is preferable to carry out ultrasonic disruption.

[0024]

Example In this example, ultrasonic disruption of microorganisms was carried out using the filtration container (filter cartridge) shown in FIG.

In FIG. 2, reference numeral 31 is an outer cylinder having a sample water inlet 32 and a filtered water outlet 33.

Inside the outer cylinder 31, one end side is cylindrical and is opened. The opening side is arranged on the filtered water outlet 33 side so that the inside of the outer cylinder is on the sample water inlet 32 side and the filtered water outlet 33 side. A partitioning filter support portion 34 is provided. The opening side of the filter support part may be arranged on the sample water 32 side.

The surface of the filter support portion 34 on the side of the sample water inlet 32 is covered with a microfilter 35 which is a microorganism trapping filter.

The filter support portion 34 is filtered by the micro filter 35 by a method such as being formed of a water permeable material or having a water permeable structure such as a metal mesh. The structure will allow water to pass through. Further, as the micro filter, one having a pore size of 0.45 μm or less is preferably used. Therefore, in the filtration container, the sample water flowing in from the sample water inlet 32 is always filtered by the microfilter 35 and then flows out from the filtered water outlet 3.

In particular, the above-mentioned microfilter is usually filtered by suction filtration, compression filtration or the like, and it does not matter that the sample water or the disrupted microorganism solution does not permeate the microfilter under normal pressure.

Therefore, in the above filtration container, after filtering the sample water by suction filtration or the like, the microbial disruption liquid is added from the sample water inflow port 32 of the outer cylinder 31 so that the space between the inner cylinder 31 and the outer cylinder 32 is increased. It is possible to store the disrupted liquid of microorganisms.

FIG. 1 shows a block diagram of the microorganism measuring apparatus according to this embodiment.

In the figure, 1 is a sample liquid tank, 14 is a sample liquid transport pipe connecting the sample liquid tank 1 and an electromagnetic valve 6, and the sample liquid transport pipe 14 has an electromagnetic valve 2, a flowmeter 3, and a sample. A pump 4 and a pressure gauge 5, which are liquid inflow means, are provided, and by driving the pump 4, the sample liquid can be injected into the filter cartridge 9 through the electromagnetic valve 6 and the injection pipe 16.

At this time, the flow rate of the sample liquid is measured by the flow meter 3.
Further, the pressure gauge 5 can measure the liquid pressure of each sample liquid.

By switching the electromagnetic valve 2, the transport of the sample liquid is stopped and the sample liquid transport pipe 1
It is also possible to open the electromagnetic valve side of No. 4 and make the inside of the sample liquid transport pipe 14 at normal pressure.

Further, 8 is a microbial crushed liquid tank, 15 is a microbial crushed liquid transport pipe connecting the microbial crushed liquid tank 8 and the electromagnetic valve 6, and the microbial crushed liquid transport pipe 15 is provided with a microbial crushed liquid injection means. A certain pump 7 is provided, and by driving this pump 7, the microbial disruption liquid can be injected into the filter cartridge 9 through the electromagnetic valve 6 and the injection pipe 16.

The filter cartridge 9 is supplied by the filter cartridge supply device 13 to the position where the sample solution and the microbial disruption solution can be supplied through the injection pipe 16.

After the filtration of the sample solution and the supply of the microbial disruption solution in this filter cartridge are completed, the sample is set in the ultrasonic disruption container by a transfer device (not shown),
Further, after the ultrasonic crushing is completed, the microbial concentrated liquid in the filter cartridge, which is set in the suction nozzle 11 and driven by the pump 12 which is the sample liquid collecting device, is set by the transfer device (not shown). It is configured to be transported to an EIA measuring device and to measure microorganisms by EIA.

In the above apparatus, the pump, electromagnetic valve and the like are controlled by the control device, and the microorganisms can be measured automatically. The control contents are shown below.

1. The sample water is supplied to the sample water tank by the water sampling device 17.

2. The transfer device is driven to set the filter cartridge 9 in the injection pipe 6 from the filter cartridge supply device 13. (Position A in FIG. 1) 3. The pump 4 is driven to feed the sample water to the filter cartridge 9 for filtration. As a result, the microorganisms remain on the filter surface as a residue.

4. The inflow amount of the sample liquid is measured by the flow meter 3, and the pump 4 is stopped when the inflow amount of the sample liquid reaches a preset integrated amount.

Alternatively, the pressure of the sample liquid is measured by the pressure gauge 5, and when the measured value reaches a preset value by the control device, it is judged that the filter is clogged and the pump 4 is stopped. You can also

At this time, the flow rate integrated value up to that point is stored.

5. The electromagnetic valve 2 is switched to open the suction side of the pump 4 to the atmosphere, and the sample water remaining in the filter cartridge is pushed out from the pump 4 to complete the filtration. At this time, the end of filtration is detected by a pressure gauge, and control is performed based on this value.

6. The sealing tube 20 is actuated to seal the tube on the outflow side of the filter cartridge 9. Examples of the sealing tube method include pressure bonding, welding and fusion.

7. After switching the electromagnetic valve 9, the pump 7 is driven to inject a fixed amount of microbial disruption liquid into the filter cartridge 9.

8. The transfer device is driven to set the filter cartridge 9 in the ultrasonic crusher 10, and the ultrasonic crusher 10 is driven (position B in FIG. 1) to crush the microorganisms in the filter cartridge 9.

9. The filter cartridge 9 is set in the suction nozzle 11 by the transfer device, the pump 12 is driven (at the position C in FIG. 1) to suck the microbial concentrated liquid, and the EI
Transport a fixed amount to the A device.

10. The concentration of microorganisms and the like are measured by the EIA measuring device. At this time, item 4. 7. Amount of filtered sample water in step 7 and item 7. The concentration ratio of the microorganism is calculated from the amount of the crushed liquid of the microorganism, and transmitted as reference data in the EIA measurement.

By carrying out the above-mentioned control with the apparatus constructed as described above, the microorganisms can be measured automatically, easily and accurately.

By repeatedly discarding the filter cartridge after performing the above operation, the concentration of microorganisms in the sample water can be measured continuously and in real time.

Therefore, especially in a wastewater treatment plant or the like, it is possible to grasp the change in water quality in real time, which greatly contributes to water quality management.

Note that item 4. In the above, the water receiving tank 18 may be provided instead of the flow meter, the water quality in the water receiving tank may be measured by the level meter 19, and the pump 4 may be controlled based on this value.

In FIG. 1, pump 4 and pump 7
However, these pumps may be provided between the electromagnetic valve 6 and the filter cartridge 9. At this time, one pump can be substituted for these pumps by switching and controlling the electromagnetic valve 6.

Further, although the crushed liquid side of microorganisms is closed and the pumps are provided on the pumping side by one pump, these pumps may be provided on the suction side of the filter cartridge 9. At this time, the pump 4 and the pump 7 may be replaced by one pump. The transfer devices may be different from each other, but the filter cartridge can be set in the injection pipe 16, the ultrasonic crushing container 10, and the suction nozzle 11 by one or two transfer devices.

[0056]

INDUSTRIAL APPLICABILITY In the present invention, the concentration of a minute amount of microorganisms can be automatically and continuously measured, and the concentration and crushing of microorganisms can be automatically and continuously performed.

Further, for example, in controlling the amount of chlorine injection in a chlorine sterilization facility, it is possible to construct a feedback system with the target number of E. coli.

[Brief description of drawings]

FIG. 1 is an explanatory diagram of a microorganism measuring apparatus according to an embodiment of the present invention.

FIG. 2 is an explanatory view of a microorganism filtration device according to an embodiment of the present invention.

FIG. 3 is an explanatory view of a microorganism filtration device according to a conventional example.

FIG. 4 is an explanatory view of a microorganism filtration device according to a conventional example.

[Explanation of symbols]

 1 ... Sample liquid tank, 2 ... Electromagnetic valve 3 ... Flowmeter 4 ... Pump 5 ... Pressure gauge 6 ... Electromagnetic valve 7 ... Pump 8 ... Microbial disruption liquid tank 9 ... Filter cartridge 10 ... Ultrasonic disrupter 11 ... Suction nozzle 12 ... Pump 13 ... Filter cartridge supply device 14 ... Sample liquid transport pipe 15 ... Microbial disruption liquid transport pipe 16 ... Injection pipe 17 ... Water sampling device 18 ... Water receiving tank 19 ... Level meter 20 ... Seal tube

Claims (4)

[Claims] 1. A sample liquid tank for storing a sample liquid, a microorganism crushed liquid tank for storing a microbial crushed liquid, and an inside of an outer cylinder provided with a sample liquid inlet and a filtered water outlet,
A filter cartridge having a microorganism-trapping filter for partitioning the inside of the outer cylinder into a sample water inlet side and a sample liquid outlet side, and a filter cartridge supply device, and a filter cartridge supplied from the filter cartridge supply device by a transfer device. Is installed at a position where the supply of the sample solution and the microbial disruption solution can be supplied, and then the sample solution obtained from the sample solution tank is introduced into the filter cartridge by the sample solution inflow means to filter the sample solution. Injecting a microbial crushed liquid obtained from the microbial crushed liquid tank by a microbial crushed liquid injecting means, after performing microbial crushing by installing the filter cartridge in which the microbial crushed liquid is injected into the microbial crusher by a transfer device, Transfer the filter cartridge to a sample liquid sampling device by a transfer device. The sample collecting device is a microorganism measuring device for measuring the microorganisms in the sample liquid by a measuring device after collecting the sample liquid that has been subjected to the microbe crushing, and the microorganism measuring device is the sample liquid inflow Means, the microorganism crushed liquid injecting means, the filter cartridge supply device, the transfer device, the microbial crusher, the sample liquid sampling device, and a control means for controlling the measuring device, thereby automating the measurement of microorganisms What has been done is a microorganism measuring device. 2. The microorganism measuring device according to claim 1, wherein the sample liquid inflow means has a liquid pressure detecting means for the sample liquid, and the control means is based on the liquid pressure obtained by the liquid pressure detecting means. A microorganism measuring device characterized by controlling the inflow of a sample liquid. 3. The microorganism measuring apparatus according to claim 1 or 2, wherein the microorganism measuring apparatus has means for detecting the inflow amount of the sample liquid and the injecting amount of the crushed microorganism liquid into the filter cartridge, and the control device is the The concentration ratio of microorganisms is calculated based on the inflow amount of the sample liquid and the injection amount of the microbial disruption liquid obtained from the detection means of the inflow amount of the sample liquid and the injection amount of the microbial disruption liquid, and is provided in advance in the microorganism measuring device. The microorganism measuring device, which outputs the concentration factor to the output unit. 4. The microorganism measuring device according to claim 1, 2 or 3, wherein the microorganism measuring device has a sealing means for sealing a sample liquid outflow portion in the filter cartridge, and the sealing means is controlled by the control device. By doing so, after the filtration of the sample liquid is completed, the sample liquid outflow portion in the filter cartridge is sealed by the sealing means, and further, an ultrasonic crusher is used as the microbial crusher, and the ultrasonic crusher is A microorganism measuring apparatus characterized by crushing microorganisms by immersing a filter cartridge and ultrasonically crushing the microorganisms.