JP4358933B2 - Prion inactivator - Google Patents
Prion inactivator Download PDFInfo
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- JP4358933B2 JP4358933B2 JP17748899A JP17748899A JP4358933B2 JP 4358933 B2 JP4358933 B2 JP 4358933B2 JP 17748899 A JP17748899 A JP 17748899A JP 17748899 A JP17748899 A JP 17748899A JP 4358933 B2 JP4358933 B2 JP 4358933B2
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- prion
- glycidol
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- inactivating agent
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Description
【0001】
【発明の属する技術分野】
本発明は、グリシドールを含んでなるプリオン不活化剤およびそれを用いてプリオンに汚染された、または汚染された可能性のある、たとえば手術具や内視鏡などの医療用具を、その材質の腐食や劣化を伴うことなく、簡便に消毒する方法に関する。
【0002】
【従来の技術】
プリオンについてはまだ不明な点が多いが、核酸の存在が証明できないことにより、DNAまたはRNAを持たない感染性蛋白といわれている。
このプリオンを病原体とするヒトの疫病としては、これまでクールー、クロイツフェルト・ヤコブ病やゲルストマン・シュトライスラー症候群などが知られており、ヒト以外ではヒツジやヤギにおけるスクレイピー、ウシにおける海綿状脳症(いわゆる狂牛病)などが知られている。医原性クロイツフェルト・ヤコブ病は、硬膜移植や、角膜移植、ヒト脳下垂体から抽出した成長ホルモンの投与や汚染された脳波電極よりヒトに伝播されたことが証明されている。
この伝播性病原体はクールーの人の脳や脊髄に大量存在することが示されており、またクロイツフェルト・ヤコブ病、ゲルストマン・シュトライスラー症候群の患者では、同様の伝播性病原体が脳、脾臓、肝臓、リンパ節、肺、脊髄、腎臓、
角膜、レンズ、脳脊髄液、血液中で検出されている。
しかしプリオンは抗体形成のような免疫応答を生じないためワクチンは存在せず、罹患の診断も極めて困難である。
【0003】
【発明が解決しようとする課題】
プリオンを不活化する方法についてはこれまで種々試みられてはいるが、この病原体は放射線、煮沸、乾熱、化学薬品(ホルマリン、ベータープロピオラクトン、アルコール、ヨード、アセトン、過マンガン酸カリ、ドデシル硫酸ナトリウム、過酸化水素、酸化エチレンガス等)を含む従来の病原微生物に対する不活化法には極めて高い耐性を有している。
高温での高濃度の鉱酸またはアルカリ溶液による処理、次亜塩素酸−アルカリ溶液による処理によりプリオンが不活化されることは知られているが、高濃度の鉱酸やアルカリ溶液は金属や有機物に対する腐蝕性が強く、次亜塩素酸−アルカリ溶液も酸化力が強いので、金属、ゴム、プラスチックなどの材質で作られている医療機器を腐蝕や劣化させ、再使用できなくすることがある。
また132℃、1時間オートクレーブにより高圧消毒する方法もあるが、大掛かりな装置を必要とし、且つ材料によっては132℃の高温に耐えないものもある。さらにエチレンオキサイド水溶液によるプリオン消毒法が提案されており(特開平09−273639,特開平10−128131)、それなりの効果が奏せられるが、エチレンオキサイドは常温でガス状のものであるので溶液を放置しておくと、ガスが揮散してエチレンオキサイドの濃度が低下し、必ずしも一定の効果を上げることができない場合がある。そこで金属や有機物に対する腐蝕性やその他の悪影響が殆どなく、高温や高圧も必要せず、簡便、確実なプリオンの消毒法が切望されている。
【0004】
【課題を解決するための手段】
本発明者らは、上記の課題を解決するために従来から用いられてきた消毒法も含め、多くの方法を検討したところ、グリシドールを含む水性溶液が簡便に調製でき、医療用機械器具を含む医療用用具に劣化や損傷を与えることなくプリオンの感染力を効果的に減衰させることができるという知見を得、その知見を基に研究を重ねて本発明を完成した。
すなわち、本発明は、
(1)グリシドールを含んでなるプリオン不活化剤、
(2)グリシドールを0.1〜30v/v%含む水性溶液である前記(1)記載のプリオン不活化剤、および
(3)医療用具を前記(1)または(2)記載のプリオン不活化剤と接触させる医療器具の消毒法、
である。
【0005】
【発明の実施の形態】
本発明のプリオン不活化剤は、主としてプリオンに汚染されたかまたはその可能性のある医療用具の消毒に用いられる。その医療用具としては病院などの医療の現場で使用されるあらゆる機械、器具、補助用具類を含む概念のもので、たとえば、静脈カテーテル、心臓カテーテルなどの各種カテーテル類、メス、鉗子、ハサミなどの手術具、気管内用チューブ、吸引チューブ、気管カニューレ、蛇腹ゴム管、気管切開用トレイなどの各種手術用器具類、胃鏡、直腸鏡など各種内視鏡や光学機器類その他人工透析具、脳用ドリル、鋸、カメラなど、再使用が望まれる医療用具をあげることができる。
本発明のグリシドールを含んでなるプリオン不活化剤は、通常グリシドールの水性溶液である。この水性溶液のために用いられる溶媒は、通常水であるが、水にエタノール等の低級アルコールやその他の親水性有機溶媒、界面活性剤などを適量、たとえば10重量%以下の割合で混合したものであってもよい。
消毒すべき対象物に接触させるグリシドール水性溶液のグリシドール濃度は、通常0.1〜30v/v%、好ましくは0.3〜10v/v%である。
消毒対象物をグリシドール含有水性溶液に接触させる時間は、通常1〜72時間、好ましくは3〜60時間、温度は通常−5〜50℃、好ましくは0〜35℃である。消毒対象物とグリシドール水性溶液との接触方法は、どのような方法でもよいが、たとえば消毒対象物をグリシドール水性溶液を収容した容器中に浸漬する方法などが挙げられる。
消毒後、医療用具はよく水洗し乾燥するだけでよい。
本発明の効果は、消毒対象物をスクレイピー感染マウス脳乳剤と接触させ、その脳乳剤が付着した消毒対象物を本発明に用いるグリシドール水性溶液の一定量中に一定時間浸漬し、消毒対象物を引き上げた後のグリシドール水性溶液を透析してグリシドールを除去し、ウエスタンブロッティング法により溶液中の抗原性の有無を調べることにより確認することができる。
【0006】
【実施例】
以下実施例により本発明をさらに詳しく説明する。
実施例1
(1)試料液の調製
カテーテルとして用いるシリコンチューブを20本用意し、それぞれのチューブの先端を1w/v%スクレイピー感染マウス脳乳剤中に浸して、脳乳剤を約100μl付着させた。
これら脳乳剤付着チューブを、0.5,1.0,3.0,5.0v/v%のグリシドール水溶液および滅菌蒸留水(対照)1mlを収容した試験管各4本(計20本)に1本ずつ浸漬し、25℃を保持しながら5,12,24および48時間時間毎に各濃度の試験管からチューブを1つづつ引き上げた。
各試験管に残ったグリシドール水溶液を、リン酸緩衝液を用いる透析に付してグリシドールを除去し試料液とした。また蒸留水にチューブを浸した対照は、チューブを引きあげ、残った液をそのまま対照液とした。
【0007】
(2)ウエスタンブロッティング法による抗原性の測定
前述の各試料液および対照液を、12.0w/v%の分離ゲルを用いて電気泳動に付し、ついで分離ゲルからポリビニリデンジフルオライド(PVDF)膜上に転写した。転写した試料をプリオン蛋白質のアミノ酸配列から設計した抗ペプチド抗体と反応させ、これに結合した抗体に、さらにこの抗体と特異的に結合し且つ酵素標識を施してある抗体とを反応させ、その抗体に標識された酵素を用いて蛍光発色させた。この発色の有無、強弱により、PVDF膜上に転写された試料の抗原性の有無、強弱を調べた。
(3)結果
前記のウエスタンブロッティング法によって得られた各試料液の抗原性の有無、
強弱を蛍光発色の有無、強弱として、ポラロイドフィルム上に感光させて下記の基準により判定した。その結果を〔表1〕にまとめた。
−:対照に比してバンドの強さに変化がない。
−〜±:僅かながらバンドの強さが減弱している。
±:明確にバンドの強さが減弱している。
±〜+:バンドは見えないが、若干反応したものが見える。
+:バンドが完全に消失している。
【0008】
【表1】
〔表1〕から明らかなように3.0v/v%以上のグリシドール水溶液では5時間の浸漬で抗原性は完全に消失した。また0.5v/v%グリシドール水溶液でも、24時間以上浸漬すると抗原性は消失する。
【0009】
【発明の効果】
本発明によれば、プリオンに汚染された、または汚染された恐れのある医療用具類を、腐蝕、劣化させることなく且つ簡便に消毒することができる。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a prion inactivating agent comprising glycidol and a medical instrument such as a surgical instrument or an endoscope that is contaminated with or possibly contaminated with prion using the prion inactivating agent. The present invention relates to a method for easily disinfecting without causing deterioration.
[0002]
[Prior art]
Although there are still many unclear points regarding prions, they are said to be infectious proteins that do not have DNA or RNA because the existence of nucleic acids cannot be proved.
Known human epidemics with prion pathogens include Kuru, Creutzfeldt-Jakob disease and Gerstmann-Strreisler syndrome. Other than humans, scrapie in sheep and goats, spongiform encephalopathy in cattle (so-called Mad cow disease) is known. It has been proven that iatrogenic Creutzfeldt-Jakob disease was transmitted to humans through dura mater transplantation, corneal transplantation, administration of growth hormone extracted from human pituitary gland and contaminated electroencephalogram electrodes.
This transmissible pathogen has been shown to be present in large amounts in the brain and spinal cord of Kuru, and in patients with Creutzfeldt-Jakob disease and Gerstman-Strreisler syndrome, similar transmissible pathogens are found in the brain, spleen, and liver. , Lymph nodes, lungs, spinal cord, kidneys,
It has been detected in the cornea, lens, cerebrospinal fluid, and blood.
However, since prions do not produce an immune response such as antibody formation, there is no vaccine and diagnosis of morbidity is extremely difficult.
[0003]
[Problems to be solved by the invention]
Although various attempts have been made to inactivate prions, the pathogens are radiation, boiling, dry heat, chemicals (formalin, beta-propiolactone, alcohol, iodo, acetone, potassium permanganate, dodecyl). It has extremely high resistance to inactivation methods for conventional pathogenic microorganisms including sodium sulfate, hydrogen peroxide, ethylene oxide gas, and the like.
It is known that prions are inactivated by treatment with high-concentration mineral acid or alkali solution at high temperature or treatment with hypochlorous acid-alkali solution, but high-concentration mineral acid or alkali solution is a metal or organic matter. Is highly corrosive, and the hypochlorous acid-alkali solution is also strong in oxidizing power, which may corrode and deteriorate medical devices made of materials such as metal, rubber and plastic, making them unusable.
There is also a method of high-pressure disinfection by autoclaving at 132 ° C. for 1 hour, but a large apparatus is required and some materials cannot withstand a high temperature of 132 ° C. Furthermore, a prion disinfection method using an aqueous ethylene oxide solution has been proposed (Japanese Patent Laid-Open Nos. 09-273369 and 10-128131), and some effects can be obtained. However, since ethylene oxide is gaseous at room temperature, a solution is used. If left untreated, the gas is volatilized and the concentration of ethylene oxide is lowered, so that a certain effect cannot always be obtained. Therefore, there is almost no corrosiveness or other adverse effect on metals and organic substances, and high temperature and high pressure are not required, and a simple and reliable prion disinfection method is desired.
[0004]
[Means for Solving the Problems]
The inventors of the present invention have studied many methods including a disinfection method conventionally used to solve the above-described problems. As a result, an aqueous solution containing glycidol can be easily prepared, and includes a medical instrument. The present inventors have obtained the knowledge that the infectivity of prions can be effectively attenuated without deteriorating or damaging the medical device, and have completed the present invention through repeated research based on the knowledge.
That is, the present invention
(1) a prion inactivating agent comprising glycidol,
(2) The prion inactivating agent according to (1), which is an aqueous solution containing 0.1 to 30 v / v% of glycidol, and (3) the prion inactivating agent according to (1) or (2) described above. Disinfection method of medical equipment to be contacted with
It is.
[0005]
DETAILED DESCRIPTION OF THE INVENTION
The prion inactivating agent of the present invention is mainly used for disinfecting medical devices contaminated with or possibly having prions. The medical device is a concept that includes all machines, instruments, and auxiliary tools used in medical settings such as hospitals, such as various catheters such as venous catheters and cardiac catheters, scalpels, forceps, and scissors. Surgical instruments, endotracheal tubes, suction tubes, tracheal cannulas, bellows rubber tubes, various surgical instruments such as tracheostomy trays, various endoscopes and optical instruments such as gastroscopes and rectoscopes, other artificial dialysis instruments, and brains Examples of medical devices that are desired to be reused include drills, saws, and cameras.
The prion inactivating agent comprising glycidol of the present invention is usually an aqueous solution of glycidol. The solvent used for this aqueous solution is usually water, but a mixture of water and a lower alcohol such as ethanol, other hydrophilic organic solvent, a surfactant, etc. in an appropriate amount, for example, a ratio of 10% by weight or less. It may be.
The glycidol concentration of the aqueous glycidol solution to be contacted with the object to be sterilized is usually 0.1 to 30 v / v%, preferably 0.3 to 10 v / v%.
The time for bringing the object to be disinfected into contact with the aqueous solution containing glycidol is usually 1 to 72 hours, preferably 3 to 60 hours, and the temperature is usually -5 to 50 ° C, preferably 0 to 35 ° C. Any method may be used for contacting the object to be disinfected with the aqueous glycidol solution, and examples thereof include a method of immersing the object to be disinfected in a container containing the aqueous glycidol solution.
After disinfection, the medical device need only be thoroughly washed and dried.
The effect of the present invention is that a disinfecting object is brought into contact with a scrapie-infected mouse brain emulsion, and the disinfecting object to which the brain emulsion is adhered is immersed in a fixed amount of glycidol aqueous solution used in the present invention for a certain period of time. The glycidol aqueous solution after being pulled up is dialyzed to remove glycidol, and it can be confirmed by examining the presence or absence of antigenicity in the solution by Western blotting.
[0006]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples.
Example 1
(1) Preparation of sample solution Twenty silicon tubes used as catheters were prepared, and the tip of each tube was immersed in a 1 w / v% scrapie-infected mouse brain emulsion to attach about 100 μl of the brain emulsion.
These brain emulsion attachment tubes were placed in 4 test tubes each containing 0.5, 1.0, 3.0, 5.0 v / v% glycidol aqueous solution and 1 ml of sterilized distilled water (control) (20 in total). One tube was dipped at a time, and the tube was pulled up one by one from the test tube of each concentration every 5, 12, 24 and 48 hours while maintaining 25 ° C.
The aqueous glycidol solution remaining in each test tube was subjected to dialysis using a phosphate buffer solution to remove glycidol, thereby preparing a sample solution. Moreover, the control which immersed the tube in distilled water pulled up the tube, and used the remaining liquid as a control liquid as it was.
[0007]
(2) Measurement of antigenicity by Western blotting The above sample solution and control solution were subjected to electrophoresis using a 12.0 w / v% separation gel, and then the polyvinylidene difluoride (PVDF) was separated from the separation gel. ) Transferred onto the membrane. The transcribed sample is reacted with an anti-peptide antibody designed from the amino acid sequence of the prion protein, and the antibody bound to this is further reacted with an antibody that specifically binds to this antibody and is labeled with an enzyme. Fluorescence was developed using an enzyme labeled with. The presence / absence and strength of the antigenicity of the sample transferred onto the PVDF membrane was examined by the presence / absence and strength of the color development.
(3) Results Presence or absence of antigenicity of each sample solution obtained by the Western blotting method,
The intensity was determined as the presence / absence or intensity of fluorescent color development, and the film was exposed to light on a polaroid film and judged according to the following criteria. The results are summarized in [Table 1].
-: There is no change in the strength of the band compared to the control.
−˜ ±: The band strength is slightly reduced.
±: The band strength is clearly reduced.
± to +: A band is not visible, but a slight reaction is visible.
+: The band has completely disappeared.
[0008]
[Table 1]
As is apparent from [Table 1], the antigenicity completely disappeared in a glycidol aqueous solution of 3.0 v / v% or more after 5 hours of immersion. Further, even in a 0.5 v / v% aqueous glycidol solution, the antigenicity disappears when immersed for 24 hours or longer.
[0009]
【The invention's effect】
According to the present invention, it is possible to easily disinfect medical devices contaminated with prions or possibly contaminated without being corroded or deteriorated.
Claims (3)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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JP17748899A JP4358933B2 (en) | 1999-06-23 | 1999-06-23 | Prion inactivator |
Applications Claiming Priority (1)
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JP17748899A JP4358933B2 (en) | 1999-06-23 | 1999-06-23 | Prion inactivator |
Publications (2)
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JP2001000512A JP2001000512A (en) | 2001-01-09 |
JP4358933B2 true JP4358933B2 (en) | 2009-11-04 |
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JP17748899A Expired - Fee Related JP4358933B2 (en) | 1999-06-23 | 1999-06-23 | Prion inactivator |
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Families Citing this family (3)
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US7803315B2 (en) * | 2001-10-05 | 2010-09-28 | American Sterilizer Company | Decontamination of surfaces contaminated with prion-infected material with gaseous oxidizing agents |
DE10203225A1 (en) | 2002-01-28 | 2003-07-31 | Weigert Chem Fab | Cleaning surgical instruments |
WO2012028196A1 (en) * | 2010-09-02 | 2012-03-08 | Ecolab Inc. | Disinfectants based on glucoprotamin with efficacy against prions |
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1999
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