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

Detection of amount of bacteria in air

Info

Publication number
JPS6016598A
JPS6016598A JP12228183A JP12228183A JPS6016598A JP S6016598 A JPS6016598 A JP S6016598A JP 12228183 A JP12228183 A JP 12228183A JP 12228183 A JP12228183 A JP 12228183A JP S6016598 A JPS6016598 A JP S6016598A
Authority
JP
Japan
Prior art keywords
air
bacteria
amount
atp
emission
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
JP12228183A
Other languages
Japanese (ja)
Inventor
Noboru Tokunaga
徳永 昇
Hiroshi Arayama
荒山 寛
Yoshito Aoyanagi
良人 青柳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hitachi Ltd
Original Assignee
Aloka Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Aloka Co Ltd filed Critical Aloka Co Ltd
Priority to JP12228183A priority Critical patent/JPS6016598A/en
Publication of JPS6016598A publication Critical patent/JPS6016598A/en
Pending legal-status Critical Current

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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

【発明の詳細な説明】 技術分野 本発明は空気中バクテリア量検出方法、特に空気中に混
入しているバクテリアの生存量を自動0′9にほぼリア
ルタイムで測定することのできる空気中バクテリア早検
出方法に関するものであり、近年の生化学ωl究にお【
プるバクテリアの飛散事故の防11等にも極めて好適な
検出方法を提供可能゛ぐある。
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.

従来技術 ?P気中バクテリア量の検出は従来において実験室的な
要請にのみこたえればよく、通常【よ適当な培地に菌を
培養して、このとぎに〈1成りる」INニー数を目視に
より数えて菌の生成数を知る方法か採られて(13す、
検出に多大の労力と11.冒111を必要とづる欠点が
あった。
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.

特に従来のJ1j養は24−48時間もの艮11゜1間
を乃り、、またこの間JH’r、地を37℃程度の一定
イ台磨に保ら続ける必要があった。従って、従来方法で
は、刻々変化する空気中のバクノリノ’tj3をリアル
タイムぐ監視することができず、例えば、近イ1の生化
学INI究その他において危険な菌が創成あるい(、U
 Fi+産され、これが外部に漏れたどきに、これを迅
速に検出して対策を講しるためには、令く役に立たない
という問題があった。
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.

更に従来の前述したJ8地法−(は、ハクーiリア吊が
多い場合に培地内でのil−成コ[に一が0いに合体混
合してしまい、正確な測定精磨がJ!lられないという
欠点を有してした。
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里!」口頭 本発明の目的は、空気中のバクテリア坦をほぼリアルタ
イムで迅速に検出し、また電気的な処理によって測定の
自動化をも可能どし、バクテリアの空気中への漏れ等に
も適切に対処できる新たな空気中バクテリア間検出方d
、を提供Jることである。
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.

発明の構成 従来の培地法に替え、バクテリア中に含まれるATP(
アデノシンミリン酸)を抽出(ッて、これに発光試薬を
加え、発光量を光電予期1B管等で電気的に検出する方
法を用い、連続的あるいは所定時間ごとの間欠的な測定
によってほぼリアルタイムで空気中バクテリア量を検出
可能とする。
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.

前記一連の作業工程は数分以内に行イつれ、従来方法に
比して箸しい時間短縮となり、生化学的に産み出された
バクテリilの空気中への拡散もその初期に発見するこ
とができ、大気汚染等を未然に防止できる。
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.

周知のように、生存バクテリi’ 1lpJ中にi、t
 ILぼ1048−10−19モルの△−1N−)が含
うIzれて、13す、△11〕の分子mは約5x 10
’でおるから、パウチリア1個中には10” −1(1
−17gの△−1’ Pが含;釘れ(−d3す、ATP
の検知能は通常1Q13g稈18であるから。
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.

103個稈磨り生存バクテリアがあ4′口;[、△−1
1)の発光J1測にて十分にバクテリアの検出がiII
能eある。
There are 103 culm polishing viable bacteria at 4'mouth; [, △-1
1) Luminescence J1 measurement was sufficient to detect bacteria.
There is noh e.

本発明にd31.ノる前記Δ−11〕の抽出はバクテリ
ア含有空気を抽出液に通りことにより1]われ、この抽
出液としでは1−リス(20m M ) ID1−△(
2mfvl)pl−17,75の煮沸液、過塩M酸(L
2N)の常温又は氷中液、hl’l酸(1,3N)の常
温又は氷中液等が6Y′)VUで゛ある。
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.

そして、双子のJ、うにして抽出されIど△11)には
発光試薬が加えられ、この発光試薬どし’−U t、t
ルシフェリン、ルシフェラーゼ等が好適’CiRす、以
下の発光反応を示ず。
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 )ルシ
フェラーゼ → AM P (、C+o Hn 07 N s P 
)+ルシフェリン(酸化型) 十〇02十PPl +光
すなわち、ATPの量に比例した560−580μの波
長の光が放出され、この発光量を光電子増倍管等にて電
気的に測定することができ、空気中のバクテリアmのリ
アルタイム測定が可能となる。
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.

従って、本発明によれば、空気中バクテリア量を連続的
に監視することによって、その気中WJ麿変化を遅れな
く知ることができ、例えば、生化学実験室内でのバクテ
リアの多聞発生あるいは実験室外への排出漏洩も未然に
確実に検知することができ、将来の生化学産業にお【ノ
る安仝性確保に寄与する屑入である。
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.

釆1目± 第1図は本発明が適用されたバクテリア14 i11’
l定装置の第1実施例である。
Figure 1 shows bacteria 14 i11' to which the present invention is applied.
This is a first embodiment of the l-determining device.

抽出器10はパフリング管12と捕4、ii!’t i
 4どを含み、イれぞれヒータ16.1 Bにょっ(抽
出液を100℃に加熱している。抽出器1oには先ず抽
出液UP 20からポンプ22にJ、リドリス、トD]
−Δ緩1チidkから成る抽出液10’0がiスリ込、
1.れ、煮沸状態にされている。
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.

次にポンプ24からLL ’Z(”;内空気′l 02
が抽出器10に吸引され、fi分数10アの空気が所定
°時間、例えは、10ヅロ111程度1汲引される。吸
引空気はバグリング管12内てのバフリングにJ、 −
) ′C抽出液ど十分に接触反応し、含有バクテリアI
からは△11)が抽出液に抽出され、イの後、−I−、
hが64ノ1気される。
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.

A T +)を抽出した抽出液はハル72 Bがらボン
728にて検出しル3oに送られ、発光試薬との発光作
用にイハされるが、このj′lS給系に、1りいC、フ
ィルタ32にて異物が除去され、また冷lJj管34に
て30℃程度に冷7i1+される。史にバルブ26は余
分な抽出液を7J) atするために制御Iす4″l(
いる。
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.

前記フィルタ332は発光作用で発光した光が後述の光
電子増倍管/10に入0・1リ−る際の障害となるホコ
リ、ゴミ等の異物が検出セル30に)lマ入り−ること
を防IF4るしのであり、木y5明の実施にあI、:つ
ては重要な役割を有する。
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.

すなわち、バクテリアは通常、空気中に、1メいC1単
独に浮遊して存Tr することは少tよく、一般にホコ
リやゴミに付着して存在4る。従って、本発明のバクテ
リア捕捉においてら、同時にゴミ等が捕集されてしまい
、この除去を行う前記フィルタ32は実際の装置には不
司欠となる。
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.

また前記冷却恰3/Iは高)#111の抽出液が検出レ
ル30に入るとき、その近傍にある光電子爵f8管40
の淘i上lにJ、る71E音の増加を防止Jるために有
益である。
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.

検出セル301.11 yOかれた抽出液には、発光−
(桑源36からルシフェリン、ルシフェラーゼから成る
発光試薬104がポンプ38にJ、って所定量加えられ
、八[1)吊り2’(わJうバクl−リi’ jjiに
比例した発光が生起される。
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.

検出しル30には光電r増倍管/IOかME接配匿され
ているので、前記発光1.1電気的にΔ!!I >iE
され、また光バ1品1忌り器42にJ、つC空気中ハク
フリアlβがデジタル顧として出ツノされる、。
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.

以上の一連の二[稈にJ:す、?1!気中バク)すj)
 5jが自動的にほぼリアルタイムで測定され、検出セ
ル30はン先浄δ髪10()をポンプ4/l(tuみ1
−げることにより洗浄され、その廃dil!t;tバル
ブ46 /)” Iら排出される。
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.

従って、本実施例にJ、れば、例えば、1j)の間隔で
空気中バクテリア吊を測定1」能とへり、気中i1i!
度の異常を迅速に検知りることかでさる。
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.

第2図に1,1、本発明を適用した測定装置の第2実施
例が示され、第1実施例と同一一部材には同一符号を付
してiJ、’明を省略づる。第2実施例は抽出器10が
若干異なり、ノズル50から出た1烏速空気流により1
fjJ1管52から吸い出された抽出液が効果的に空気
と接触し、この間にΔ−[1〕の抽出が71われること
を特徴とりる。
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.

発明の詳細 な説明しIc J:うに、本発明によれば、バクテリア
吊の検出のために該バクテリア吊とり・1応関係にある
Δ−11つを抽出し、これに発光試薬を加え(発光作用
を生起させ、次に光電変換による電気的1、、> 4.
=y、 、Nの検出を?7つ゛(いるので、はぼリアル
タイムで遅れなく迅速にバクアリア印の測定を行うこと
が可01:となり、イ1ユ化学分野にi13い(広範1
111に1史用することができる。
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.

【図面の簡単な説明】[Brief explanation of the drawing]

第1図【J本発明が適用された1llll定装置の第1
実施例を承り概略構成図、 第2図は本発明が適用された測定装置の第2実施例を承
り概略構成図(・ある。 ’+ 00 ・・・ 抽出液 102 ・・・ 被検空気 104 ・・・ 発光試薬。
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)

【特許請求の範囲】[Claims] (1)被検空気を所定時間抽出液中に通して空気中バク
テリア部に3=1応した△TPを抽出し、K哀AT’ 
P抽出液に発光試薬を加えて発光を起こさしこの発光h
lを電気的に旧制し、はばりj′ルタイムにて空気中の
バクテリア吊を測定することのて′きる空気中バクテリ
ア量検出方法。
(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.
JP12228183A 1983-07-07 1983-07-07 Detection of amount of bacteria in air Pending JPS6016598A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP12228183A JPS6016598A (en) 1983-07-07 1983-07-07 Detection of amount of bacteria in air

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP12228183A JPS6016598A (en) 1983-07-07 1983-07-07 Detection of amount of bacteria in air

Publications (1)

Publication Number Publication Date
JPS6016598A true JPS6016598A (en) 1985-01-28

Family

ID=14832074

Family Applications (1)

Application Number Title Priority Date Filing Date
JP12228183A Pending JPS6016598A (en) 1983-07-07 1983-07-07 Detection of amount of bacteria in air

Country Status (1)

Country Link
JP (1) JPS6016598A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0738121U (en) * 1993-12-22 1995-07-14 有限会社いわしや宏仁堂 Medical underwear
US5773710A (en) * 1994-03-18 1998-06-30 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Cellular material detection apparatus and method

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5768795A (en) * 1980-10-14 1982-04-27 Dainatetsuku Ag Detection of bacteria in urine

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5768795A (en) * 1980-10-14 1982-04-27 Dainatetsuku Ag Detection of bacteria in urine

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH0738121U (en) * 1993-12-22 1995-07-14 有限会社いわしや宏仁堂 Medical underwear
US5773710A (en) * 1994-03-18 1998-06-30 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland Cellular material detection apparatus and method
AU699575B2 (en) * 1994-03-18 1998-12-10 Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Ireland, The Cellular material detection apparatus and method
EP0789778B1 (en) * 1994-03-18 2001-05-30 The Secretary Of State For Defence In Her Britannic Majesty's Government Of The United Kingdom Of Great Britain And Northern Cellular material detection apparatus and method

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