JP2000078999A - Method and apparatus for detecting microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for detecting microorganisms, capable of promptly performing the detection of the microorganisms such as pathogenic bacteria, and further to provide an apparatus therefor. SOLUTION: Pathogenic bacteria in a test sample 1 is subjected to a dyeing treatment by an antibody-dyeing or the like, and the sample 1 is photographed to provide image data. The obtained image data are subjected to an image treatment 9 for removing the image without the one obtained by the dyeing treatment from the image data by color distinction. The treated image data is subjected to a three-dimensional treatment 10 for converting the individual shape in the image data to a three-dimensional data. The obtained three- dimensional data is subjected to a collation treatment 12 for collating the obtained three-dimensional data with the known three-dimensional data of the pathogenic bacteria in a recording means 11, and the calculation treatment of the pathogenic bacteria is carried out.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for detecting microorganisms present in a test sample and an apparatus used for carrying out the method. It can be used for sanitary management such as detection of the presence or absence of a pathogen, detection of the type of pathogenic bacterium, detection of the presence or absence of the pathogenic bacterium when it is present, and detection of the number of pathogenic bacteria.

[0002]

2. Description of the Related Art A conventional method for testing microorganisms such as bacteria involves culturing microorganisms in a culture site and examining the microorganisms in colonies. In addition, the conventional method for examining the number of microorganisms has been an artificial method using a microscope, a counter, and the like.

[0003]

However, according to these conventional methods, there is a problem that it takes a long time until an inspection result is obtained. In particular, in microbiological testing of foods produced in food production factories, it is required that test results be obtained before the food is shipped, and prompt inspection is required. Not previously offered.

[0004] In addition, conventional microbiological tests on foods have been performed on common Escherichia coli and live bacteria, but with the diversification of foods, various foods such as vacuum-packed foods have been put on the market. ing. Among these foods are certain pathogenic bacteria such as Salmonella, Clostridium botulinum, Vibrio parahaemolyticus, Campylobacter, small spherical virus, O-1.
There is a need for the development of a microorganism detection technology that can easily detect such specific pathogens in a short time and utilize the data for a food production management system in factories and the like. It was started.

[0005] An object of the present invention is to provide a method for detecting microorganisms which enables rapid detection of microorganisms such as pathogenic bacteria and can be effectively used for sanitary control in various places including food production factories. And its apparatus.

[0006]

According to the method for detecting microorganisms of the present invention, after performing a staining treatment for staining microorganisms contained in a test sample, the test sample is photographed to create image data, Performing image processing for removing images other than those stained in the staining process by color discrimination from the image data, and thereafter, detecting the microorganisms from the image data subjected to the image processing. is there.

According to this method, microorganisms in the test sample are stained, and image processing for removing images other than those stained from image data obtained by photographing the test sample by color discrimination is performed. Microorganisms can be easily detected even if there are a large number of microorganisms other than microorganisms. Even if not only microorganisms but also specific substances other than microorganisms are stained, the number of individuals included in image data subjected to image processing by color discrimination is included in image data obtained by photographing the test sample. However, the number of microorganisms is smaller than the number of individuals, so that microorganisms can be easily detected.

[0008] For this reason, it is possible to achieve quick and reliable detection of microorganisms.

In the above-mentioned present invention, microorganisms may be detected immediately after performing image processing for removing images other than those stained from image data obtained by photographing a test sample. After performing the conversion, the shape of the microorganism in the image data is converted into three-dimensional data, and the three-dimensional data and the data in the microorganism image recording means that records the three-dimensional data already known for the microorganism are converted. The collation may be performed, and the microorganism may be detected by the collation.

According to the latter, a specific microorganism which is difficult to distinguish from other microorganisms in two-dimensional data based on image data can be reliably distinguished from other microorganisms. For this reason, when microorganisms are present in the test material, the three-dimensional data of a plurality of types of microorganisms are recorded in the microorganism image recording means, so that the types of microorganisms present in the test material can be determined. It can be detected and the number of the microorganism can be accurately counted.

In the above, the staining treatment for staining the microorganism may be performed by dyeing the cell nucleus of the microorganism with a dye, or when the microorganism is a pathogenic bacterium, may be performed by antibody staining.

[0012] Image processing for removing images other than those stained from the image data obtained by photographing the test sample can be performed by computer technology, and it is not possible to convert the shape of microorganisms from two-dimensional data to three-dimensional data. The three-dimensional data of the microorganism and the data in the microorganism image recording means can also be collated by the computer technology using the image sensing technology of the computer. The counting of microorganisms can be performed by computer pattern recognition technology.

[0013] The photographing of the inspection material to create the image data may be performed only once for one inspection material, or may be performed a plurality of times continuously.

According to the latter, when a microorganism is present in the test data, if the microorganism is alive, the three-dimensional data on the microorganism in the image data changes with the movement of the microorganism, and the microorganism is dead. If this three-dimensional data does not change, it will be possible to detect the life and death of microorganisms.

The three-dimensional data of the microorganisms to be recorded in the microorganism image recording means may be only the data of the microorganism alone. However, the data of the microorganism alone and the plurality of microorganisms are linked or aggregated. It is preferable to keep both of the data.

By employing the latter, it is possible to detect even microorganisms that may be in a chain or nodule shape, and the types of microorganisms that can be detected can be expanded.

The above detection method according to the present invention can be applied to any microorganism, one of which is a pathogenic bacterium. Also,
This pathogenic bacterium may be present in foods and related products, and may be present in foods and related products, such as factory water, factory wastewater, domestic water, domestic wastewater, pools, live fish tanks, water bodies, lakes and marshes, and sea areas. You may do it.

The term "food" as used herein refers to the food itself, and "related food" refers to food additive water, food wash water, food material wash water, and the like. Therefore, when using microorganisms as food pathogens, the test sample may be obtained from the food itself, food additive water or food wash water,
It may be obtained from food material washing water or the like.

When the detection method according to the present invention is applied to pathogenic bacteria of food produced by a food production device, the detection data obtained by detecting the pathogenic bacteria may be sent to a control device of the food production device. According to this, the food production device can be controlled with the detection data regarding the counting of pathogenic bacteria and the like.

Further, the detection of pathogenic bacteria may be carried out randomly only when required for food produced by the food production apparatus, or may be carried out periodically. If done regularly, food production can be carried out under sanitary control.

Controlling a food production apparatus with detection data means that when a pathogenic bacterium is detected from a test sample or when a pathogenic bacterium of a certain number or more is detected, the apparatus producing the food is controlled. To stop the operation, to increase the heating temperature by operating a device for heating the food material or the food itself, and to operate a device for disposing of the food material or the food itself, etc. Alternatively, a sound or light alarm device attached to the food production device may be activated to notify a worker engaged in the food production device.

The apparatus for detecting microorganisms according to the present invention comprises an image data recording shape data of microorganisms contained in a test sample and a microorganism image recording means recording shape data already known about the microorganisms. It is characterized by having matching means for matching with data.

According to this detection device, the microorganism to be detected present in the test sample can be accurately determined based on the data in the microorganism image recording means, and the determination can be performed quickly, and the determination result can be obtained in a short time. You will be able to get.

When the detection device is a computer, the microorganism image recording means may be a hard disk, or may be a magnetic disk or an optical disk to which recording data is input from an input device of the computer.

In the detection device according to the present invention, the shape data of the microorganism may be two-dimensional data or three-dimensional data.

Further, the detection device according to the present invention can be used for detecting various microorganisms including pathogenic bacteria, and when used for detection of pathogenic bacteria in food produced by a food production device, the detection device is used. It may be a part of a food production device.

[0027]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS One embodiment of the present invention will be described below with reference to the drawings. FIG. 1 shows a flow of a procedure for implementing the detection method according to the present embodiment, and this flow is for a pathogenic bacterium of a food produced by a food production device.

First, an inspection sample 1 for food produced by a food production device is prepared. When the test sample is the food itself, the test sample 1 is prepared by crushing the sample that has been peeled or excised from the part to be tested, and the test sample is formed of water such as food additive water, food wash water, or food material wash water. If it is food-related water, make it with a composite sample.

Next, the pathogenic bacteria present in the test sample 1 are stained by antibody staining or dye staining, and the test sample 1 is housed in a test cell to produce a pathogenic bacterial cell 2. Thereafter, the pathogen cell test cell 2 is photographed by a CCD camera device while being illuminated by the illumination device 3, and image data of the test sample 1 is created. The illuminating device 3 used at this time can irradiate a large amount of such a specific wavelength of light, as long as the staining material used in the staining treatment of the pathogenic bacterium is particularly responsive to light of a specific wavelength such as ultraviolet light. .

The image data obtained by the above photographing is sent to the computer 4. The hardware of the computer 4 is shown in FIG. The computer 4 includes an arithmetic processing unit 5
, An input device 6, a recording device 7, and an output device 8.

The input device 6 includes an input section for inputting data recorded on external recording means such as a keyboard and a magnetic disk, and an input section for inputting the image data. The recording device 7 stores a basic program for performing predetermined processing in the arithmetic processing device 5 in accordance with an instruction signal from a keyboard of the input device 6, a processing program necessary for performing each processing shown in FIG. 1, and the like. Has been
Further, it has a recording unit for recording data input from the input device 6. The output device 8 includes a display unit that displays a result of the arithmetic processing performed by the arithmetic processing device 5 on a screen,
A printer unit for printing and displaying, and an output unit for outputting and recording the arithmetic processing result data to external recording means such as a magnetic disk, and further, an output for outputting the arithmetic processing result data to a control device of the food production apparatus. Output device 8
It is provided in.

In FIG. 1, image data of an inspection sample 1 obtained by photographing with a CCD camera device is input from an input device 6 to a computer 4, and the image data is
First, an image processing 9 for removing video data other than the image data stained in the staining process from the image data as noise data is performed. The image processing 9 is performed in the arithmetic processing unit 5 by an unnecessary image removal processing program that leaves only the colors that are recorded in the recording device 7 and are colored in a specific wavelength range by color discrimination.

As a result, the image data of the test sample 1 obtained by the above photographing becomes data of an extremely small number of individuals.

Next, a three-dimensional processing 10 for converting the shape of the individual in the image data obtained in the image processing 9 from two-dimensional data to three-dimensional data is performed. This three-dimensional processing 10
Is recorded in the recording device 7 and is performed in the arithmetic processing device 5 by a processing program of image sensing which determines the height based on a difference in partial density of the individual.

As a result, even if the two-dimensional shape of a specific pathogen to be detected is similar to the two-dimensional shape of another pathogen, the difference can be clarified by converting it into three-dimensional data. Even if the pathogens other than the pathogens contained in the test sample 1 at the time of performing the staining process are stained and remain in the image data after the image processing 9, the difference from the specific pathogens is clearly identified. Can be changed.

Next, the three-dimensional data of the individual in the image data obtained in the three-dimensional processing 10 and the input device 6 of the computer 4 are recorded in the recording device 7, and the specific pathogen is already known. A collation process 12 for collating the three-dimensional shape data with the data in the pathogen image recording means 11 that records the three-dimensional data is performed. This collation processing 12 is performed by a collation processing program recorded in the recording device 7.
This is performed in the arithmetic processing unit 5.

Thus, the specific pathogenic bacterium can be distinguished from other pathogenic germs and individuals other than the pathogenic bacterium. Thereafter, the image data obtained in the three-dimensional processing 10 is used to leave the specific pathogenic bacterium alone. The processing for removing the pathogenic bacteria and the individual is performed by the processing program recorded in the recording device 7.

Then, a counting process 13 for counting the specific pathogenic bacteria from the image data thus obtained is performed. This counting process 13 is performed in the arithmetic processing device 5 by a pattern recognition counting process program recorded in the recording device 7.

As a result, whether or not a specific pathogen exists in the test sample 1, and if so, the number of individuals is counted.

The data resulting from the image processing 9, the three-dimensional processing 10, the collation processing 12, and the counting processing 13 are output to the output device 8
The display data is displayed on the display unit 8A, and the detection data obtained in the counting process 13 is converted into a printed matter 14 by the printer unit of the output device 8, and the detection data is stored in the recording unit of the output device 8 as necessary. The data is output from the recording output unit to recording means 15 such as a magnetic disk and recorded.

Further, the detection data obtained in the counting process 13 is sent to a control device of a food production apparatus on which the test sample 1 is based, and is used as a signal for controlling the production apparatus. You. That is, according to the detection data,
When a specific pathogen is detected in food that must not be present, or when a specific pathogen is detected in a food that must not have a certain number or more of the population, the number of individuals that is equal to or higher than the certain value is detected The detection data is used for performing a measure such as stopping the operation of the food production apparatus.

Thus, it becomes possible to automatically control the operation of the food production apparatus.

Further, by performing the flow for counting pathogenic bacteria shown in FIG. 1 periodically during the production of food, the production of food can be performed under sanitary control.

Further, the pathogen image recording means 1 shown in FIG.
If data on a plurality of pathogens is recorded in 1, it is possible to detect what kind of pathogen was present in the test material 1, and the type of pathogen can be detected.

Furthermore, if the photographing by the CCD camera device shown in FIG. 1 is continuously performed a plurality of times and the image data obtained by the photographing is continuously generated, live pathogens exist in the inspection material 1. In this case, at the stage of the image processing 9 or the stage of the three-dimensional processing 10, the data on the pathogen changes with the movement of the pathogen, so that it is possible to determine that the pathogen is alive. On the other hand, when the pathogenic bacterium is dead, the data on the pathogenic bacterium does not change. It is understood that control such as stopping the operation of the food production apparatus is unnecessary, and the operation control of the food production apparatus can be performed in such a manner.

[0046]

According to the present invention, the effect that the detection of microorganisms can be performed quickly can be obtained.

[Brief description of the drawings]

FIG. 1 is a flowchart showing a detection method according to an embodiment of the present invention.

FIG. 2 is a diagram of hardware of the computer shown in FIG.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Inspection sample 3 Lighting device 4 Computer 9 Image processing 10 Three-dimensional processing 11 Pathogen image recording means 12 Verification processing 13 Counting processing

Claims (7)

[Claims] After performing a staining process for staining a microorganism contained in a test sample, the test sample is photographed to create image data, and the image data is stained in the staining process by color discrimination. A method for detecting microorganisms, comprising: performing image processing for removing a video other than a subject, and thereafter detecting the microorganism from image data on which the image processing has been performed. 2. The method for detecting microorganisms according to claim 1, wherein after performing the image processing, the shape of the microorganisms in the image data is converted into three-dimensional data, and the three-dimensional data and the microorganisms are converted. A method for detecting microorganisms, comprising: comparing the known three-dimensional data with data in a microorganism image recording means that records the three-dimensional data, and detecting the microorganism by the comparison. 3. The method for detecting a microorganism according to claim 1, wherein the test sample is obtained from a food or a related substance thereof, and the microorganism is a pathogen of the food. A method for detecting microorganisms. 4. The method for detecting microorganisms according to claim 3, wherein after the detection of the pathogenic bacteria, the detection data is sent to a control device of the food production device, and the production device is controlled. A method for detecting microorganisms. 5. The method for detecting a microorganism according to claim 4, wherein the detection of the pathogenic bacterium is periodically performed on food produced by the production device or a related product thereof. . 6. A collating means for collating image data in which shape data of a microorganism contained in a test sample is recorded with data in a microorganism image recording means in which shape data already known about the microorganism is recorded. An apparatus for detecting microorganisms, comprising: 7. An apparatus for detecting microorganisms according to claim 6, wherein the test sample is obtained from food or a related substance thereof, and the microorganism is a pathogen of food. apparatus.