JPS6293634A - Microbe counter - Google Patents

Abstract

PURPOSE:To make it possible to rapidly measure the number of microbes in air without requiring culture, by collecting microbes contained in sample air and generating luminescent reaction in extracted ATP to detect light. CONSTITUTION:When an air pump 6 is operated, air flows through a sample air line 5 from an sample air suction port 1 and is filtered by the membrane filter (MF)11 grasped by a flange 4 to be exhausted through a sample air exhaust port 7. MF11 moves to a chemical liquid tank 14 at every one rotation of a roller 8 and adenosine triphosphate ATP in a microbial cell collected on MF11 is sufficiently contacted with a tris-EDTA buffer solution 15 and extracted. Extracted ATP advances onto the chemical solution tank 18 and reacted with a luciferin/luciferase reagent to generate bio-luminescent reaction. The light emitted at this time is condensed to a light condensing box and detected by a photomultiplier tube 23 through a condensing lens 22 and subjected to signal processing by a signal processing part 24 to quantity ATP.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a microorganism counter for measuring microorganisms in the air.

Conventional technology Traditionally, when measuring the number of microorganisms in the air, microorganisms in the air are placed in an appropriate medium using the drop method or impact method, the medium is cultured under appropriate conditions for a certain period of time, and the number of colonies generated is counted. It was measured as the number of microorganisms.

Problems to be Solved by the Invention As described above, in the conventional method for measuring microorganisms in the air, the culture process is unavoidable, so the number of microorganisms can be determined for about 24 to 48 hours for bacteria and 2 to 7 hours for fungi. It took about a day. For this reason, even if microorganisms that exceed safety standards occur in places where microbial control is required, such as food factories and pharmaceutical factories, appropriate measures are not taken immediately, resulting in food poisoning and product defects. It was a problem.

In view of the above-mentioned problems, the present invention aims to provide a microorganism counter that can quickly measure the number of microorganisms in the air.

Means for Solving the Problem In order to solve this problem, the microorganism counter of the present invention includes means for sucking air as sample air and collecting microorganisms contained in the sample air, and a means for collecting microorganisms contained in the sample air. It has a means for extracting ATP from the collected microorganisms, a means for causing a luminescence reaction in the ATP extracted by the ATP extraction means, and a means for detecting light obtained as a result of the luminescence reaction. With this configuration, when measuring the number of microorganisms in the air,
Microorganisms present in the sample air are collected by the collection means, and ATP in the body is extracted by the ATP extraction means. The extracted ATP undergoes a bioluminescence reaction with a luciferin/luciferase reagent under certain conditions to emit light. By detecting this light, the number of microorganisms in the sample air can be determined.

EXAMPLE Hereinafter, an example of the present invention will be explained based on the drawings. A sample air suction port 1, a sample air flow rate control valve 2, a flexible joint 3, a flange 4, an air pump 6, and a sample air exhaust lower are attached to the sample air line 6. A roller 8 having a protrusion 8a provided in contact with the pull air line 5 is provided on the side of the sample air suction port 1 and the sample air exhaust port γ, respectively.

A hydrophilic membrane filter 11 with a pore diameter of 0.2 μm is placed between the rollers 9 and 10, and the roller 1o is connected to a motor 12 by a belt 13. This membrane filter 11 has a flange 4 in the middle.
It is inserted inside. The men's filter 11 that has exited the flange 4 reaches the first chemical tank 14 .
Inside is Tris (O, OS mol hydroxymethylamine methane) EDTA (ethylenediaminetetraacetic acid) buffer solution 16
When the roller 16 contacts the membrane filter (hereinafter referred to as MF) 11, Tris-EDT is stored.
Thoroughly impregnate MFll with A buffer solution 16. This first
A heater 17 is provided at the bottom of the chemical tank 14 . 1st
The MFll that has exited the chemical solution tank 14 reaches the second chemical solution tank 18, and a bath solution 19 containing luciferin/luciferase test i and magnesium ions is stored in the second chemical solution tank 18.
By contacting the MFll while the roller 20 rotates, the MFll can be sufficiently impregnated with the solution 19 containing the luciferin/luciferase reagent and magnesium ions. MF 1 that has exited this second chemical tank 18
1. A light collecting box 21 is provided to collect the light generated by the luminescence reaction on the top of the light collecting lens, and a light collecting lens is provided on the upper part of the light collecting box 21, and the light obtained by the collecting lens 22 is detected and electrically generated. Photomultiplier tube 23 that outputs a signal
A signal processing section 24 for processing signals from the photomultiplier tube 23 is provided.

The operation of the microorganism counter configured as described above will be explained below.

When the air pump 6 is operated, air flows through the sample air line 6 from the sample air suction port 1 while being adjusted in flow rate by the sample air flow control valve 2, is filtered by the MFll sandwiched between the flanges 4, and the sample air is Exhausted from the exhaust lower. The pore size of MFll is 0.2 S chron, which makes it possible to almost completely capture microorganisms floating in the air.

When the roller 8 rotates and its protrusion 8a contacts the sample airline 6, the 7 langes 4
1 and the air pump 6 stops.

While 7 lunge 4 is away from MFll, motor 1
2 rotates, which rotates the roller 1o via the belt 13, thereby moving the MFll toward the first chemical tank 14. When the protrusion 8 & of the roller 8 separates from the sample airline 6, the movement of the MFll stops and again.
The MFll is inserted into the 7-lunge 4, and the air pump 6 is operated to continue p-passage. On the other hand, the moved MFll is first set so as to stop on the first chemical liquid tank 14. On top of the MFll which is sandwiched between flanges 4 and passes air,
Almost 100% of the microorganisms in the air have been collected, and ATP (adenosine myric acid) within the microorganisms can be extracted by sufficiently contacting them with the Tris-EDTA buffer 16. Tris EDTA buffer 16 is thoroughly impregnated by capillary development of MFll. Also, Tris EDTA buffer 1
6 In order to keep the activity level of the mosquitoes high, the heater 17
It is heated to 0°C. The extracted ATP then advances onto the second chemical solution W118 and undergoes a luminescence reaction. That is, when the solution 19 containing the luciferin/luciferase reagent and magnesium ions reacts with ATP in the presence of molecular oxygen, a reaction accompanied by a luminescence phenomenon (luminescence reaction or pyoluminescence reaction) occurs. This is because ATP is dephosphorylated to produce AMP and birophosphate, and luciferin, luciferase, and AM
Combines with P and 5. Molecular oxygen reacts with this complex,
Oxyluciferin and carbon dioxide gas are produced and AMP is separated. At this time, photons are emitted and the peak is 5
It is said that 62 nm light emission occurs.

The reaction formula is as follows.

The light emitted at this time is collected by a condensing box 21, detected by a photomultiplier tube 23 via a condensing lens 22, and then sent to a signal processing unit 2.
If signal processing is performed in step 4, the amount of ATP can be quantified.

Effects of the Invention As described above, according to the present invention, there is provided a means for collecting microorganisms contained in sample air, and a means for collecting ATP from the collected microorganisms.
By having a means for extracting , a means for causing a luminescence reaction, and a means for detecting light, the number of microorganisms in the air can be measured without the need for culturing, so the number of microorganisms can be quickly determined.

[Brief explanation of drawings]

The figure is a configuration diagram showing a microorganism counter according to an embodiment of the present invention. 4...7 lunge, 6...air pump,
11...Membrane filter-114...
...First chemical tank, 16...Tris EDTA buffer, 16, 20...Roller, 18...
Second chemical tank, 19... Solution containing luciferin/luciferase reagent and magnesium ion, 23...
...Photomultiplier tube.

Claims (1)

[Claims] a means for sucking air as sample air and collecting microorganisms contained in the sample air; a means for extracting ATP (adenosine myric acid) from the microorganisms collected by the collecting means; and extraction by the ATP extracting means. AT
A microorganism counter comprising means for causing a luminescence reaction in P, and means for detecting light obtained as a result of the luminescence reaction.