JPS6016598A - Detection of amount of bacteria in air - Google Patents

Abstract

PURPOSE:To detect an amount of bacteria in air in a real time rapidly, by passing air to be tested through an extracting solution for a given time to extract ATP (adenosinemyrinic acid), adding a coloring reagent to it, detecting luminous energy of emission electrically. CONSTITUTION:The extracting solution 100 consisting of tris EDTA buffer solution is sent to the extraction device 10 consisting of the bubbling tube 12 and the collecting well 14, it is made into a boiling state, air in the chamber 102 is sucked by the pump 24, the sucked air is brought into contact with the extracting solution sufficiently, and ATP is extracted from bacteria contained in air. Impurities are removed from the solution having extracted ATP by the filter 32, cooled by the cooler 34, sent to the detection cell 30. A given amount of a coloring reagent (e.g., luciferin, or luciferase) 104 is added to it, emission of an amount of ATP, namely emission proportional to an amount of bacteria is caused, the emission is measured electrically by the photoelectron amplifying tube 40, and the amount of bacteria is outputted as a digital value by the calculator 42 for luminous energy.

Description

DETAILED DESCRIPTION OF THE INVENTION Technical Field The present invention relates to a method for detecting the amount of bacteria in the air, and in particular, an early detection method for airborne bacteria that can automatically measure the amount of viable bacteria in the air almost in real time. It is related to the method and has been used in recent biochemical ωl research.
It is also possible to provide a detection method that is extremely suitable for preventing bacterial scattering accidents.

Conventional technology? Detection of the amount of P airborne bacteria has conventionally been carried out only in response to laboratory requests, and is usually carried out by cultivating bacteria in a suitable medium and then visually counting the number of IN knees that reach 1. A method was adopted to determine the number of bacteria produced (13th,
11. It takes a lot of effort to detect. It had the drawback of requiring 111 steps.

In particular, conventional J1J cultivation requires 24 to 48 hours of 11°C, and during this period it is necessary to maintain the JH'r soil at a constant temperature of about 37°C. Therefore, with conventional methods, it is not possible to monitor in real time the ever-changing Bacterium in the air, and for example, dangerous bacteria may be created or
There is a problem in that it is of no use in quickly detecting and taking countermeasures when Fi+ is produced and leaks to the outside.

Furthermore, in the conventional J8 soil method mentioned above, when there is a lot of filtration, the filtrate in the medium may be mixed and mixed, making accurate measurement difficult. It had the disadvantage that it could not be used.

1 ri! The purpose of the present invention is to quickly detect bacteria in the air in almost real time, and also to automate the measurement using electrical processing, and to appropriately deal with the leakage of bacteria into the air. A new way to detect bacteria in the air d
, to provide the following.

Structure of the invention Instead of the conventional culture medium method, ATP (
Adenosine myphosphate) is extracted, a luminescent reagent is added to it, and the amount of luminescence is electrically detected using a photoelectric detector 1B tube, etc., and the measurement is carried out continuously or intermittently at predetermined time intervals in almost real time. The amount of bacteria in the air can be detected.

The series of steps described above can be completed within a few minutes, which is a significant time reduction compared to conventional methods, and the diffusion of biochemically produced bacteria into the air can be detected at an early stage. It is possible to prevent air pollution, etc.

As is well known, in living bacteria i' 1lpJ, i, t
IL contains 1048-10-19 moles of △-1N-), and the molecule m of 13s, △11] is about 5x 10
', so one pouchilia contains 10"-1(1
-17g of △-1'P included; Nail (-d3S, ATP
The detection capacity is usually 1Q13g culm 18.

There are 103 culm polishing viable bacteria at 4'mouth; [, △-1
1) Luminescence J1 measurement was sufficient to detect bacteria.
There is noh e.

d31. The above Δ-11] is extracted by passing bacteria-containing air through the extract, and this extract has 1-lith (20 m M ) ID1-Δ(
2mfvl) boiling liquid of pl-17,75, persalt M acid (L
A solution of 2N) at room temperature or in ice, a solution of hl'l acid (1,3N) at room temperature or in ice, etc. are 6Y')VU.

Then, a luminescent reagent is added to the twin J, which is extracted by
Luciferin, luciferase, etc. are suitable for CiR, but do not exhibit the following luminescent reaction.

ATP (CID HI3013 N5 P3) Luciferase → AMP (, C+o Hn 07 N s P
) + Luciferin (oxidized type) 10020PPl + Light, that is, light with a wavelength of 560-580 μ that is proportional to the amount of ATP is emitted, and the amount of light emitted is measured electrically with a photomultiplier tube, etc. This enables real-time measurement of bacteria m in the air.

Therefore, according to the present invention, by continuously monitoring the amount of bacteria in the air, it is possible to know without delay changes in the amount of bacteria in the air. It is possible to reliably detect any leakage of waste into the biochemical industry in the future, contributing to greater safety in the biochemical industry.

Figure 1 shows bacteria 14 i11' to which the present invention is applied.
This is a first embodiment of the l-determining device.

The extractor 10 has a puffing tube 12 and a trap 4, ii! 't i
Each heater 16.1 B (heats the extract to 100°C. In the extractor 1o, first, extract liquid UP 20 is sent to pump 22, J, lidoris, and D).
- 10'0 of extract consisting of Δ 1 idk is added,
1. and is brought to a boil.

Next, from the pump 24 LL 'Z(''; internal air 'l 02
is drawn into the extractor 10, and air of fi fraction 10 is pumped out for a predetermined time, for example, about 10 111. The suction air is sent to the buff ring inside the bag ring pipe 12.
) 'C extract was subjected to sufficient contact reaction, and contained bacteria I
△11) is extracted into the extract, and after a, -I-,
h is 64 no 1 ki.

The extract obtained by extracting A T , foreign matter is removed by a filter 32, and cooled to about 30° C.7i1+ by a cold lJj pipe 34. In addition, valve 26 is controlled to remove excess extract liquid (7J).
There is.

The filter 332 prevents foreign matter such as dust and dirt from entering the detection cell 30, which would be an obstacle when the light emitted by the light emitting action enters the photomultiplier tube/10 (to be described later). It is a defense against IF4 and has an important role in the implementation of tree Y5 light.

That is, bacteria usually exist in the air, floating on a single layer of C1, but generally attached to dust and dirt. Therefore, in the bacteria trapping of the present invention, dust and the like are also collected at the same time, and the filter 32 that removes this is indispensable in an actual device.

In addition, the cooling rate 3/I is high) When the #111 extract enters the detection rail 30, the photoelectronic F8 tube 40 near it
It is beneficial to prevent an increase in the number of noises that may occur during the process.

Detection cell 301.11 yO extracted liquid has luminescence-
(A predetermined amount of a luminescent reagent 104 consisting of luciferin and luciferase is added to the pump 38 from the mulberry source 36, and luminescence proportional to the amount of light emitted is generated. be done.

Since the photomultiplier tube/IO or ME is connected to the detector 30, the above-mentioned light emission 1.1 electrically Δ! ! I>iE
In addition, J and C airborne Hakufuria lβ are added to the light bar 1 product 1 repellent device 42 as a digital agent.

The above series of two [culm J: Su...? 1! Air baku)suj)
5j is automatically measured in almost real time, and the detection cell 30 is pumped 4/l (tu mi 1
- Cleaned by removing the waste dil! t; t valve 46 /)''I is discharged.

Therefore, in this example, if J, for example, 1j), the airborne bacterial suspension is measured at an interval of 1", then the airborne i1i!
The key is to quickly detect abnormalities.

FIG. 2 shows a second embodiment of a measuring device to which the present invention is applied, in which the same members as in the first embodiment are denoted by the same reference numerals, and ``iJ'' is omitted. In the second embodiment, the extractor 10 is slightly different, and the 1-coast air flow from the nozzle 50 produces 1
It is characterized in that the extract sucked out from the fjJ1 tube 52 effectively comes into contact with air, during which time 71 extraction of Δ-[1] is carried out.

Detailed Description of the Invention Ic J: According to the present invention, in order to detect bacterial suspension, Δ-11, which is related to bacterial suspension, is extracted, and a luminescent reagent is added thereto (luminescent 1, >4.
Detection of =y, ,N? Since there are 7 ゛, it is possible to quickly measure the bacterium mark in real time without any delay, making it possible to quickly measure the bacterium mark in the field of chemistry.
It can be used once in 111.

[Brief explanation of the drawing]

FIG.
Figure 2 is a schematic diagram of the second embodiment of the measuring device to which the present invention is applied. ... Luminescent reagent.

Claims (1)

[Claims] (1) Pass the test air through the extraction solution for a predetermined period of time to extract △TP that corresponds to 3=1 to the bacterial part in the air.
A luminescence reagent is added to the P extract to cause luminescence, and this luminescence h
A method for detecting the amount of bacteria in the air that electrically controls the amount of bacteria in the air and can measure the amount of bacteria in the air in real time.