Stock material or miscellaneous articles – Web or sheet containing structurally defined element or... – Adhesive outermost layer
Reexamination Certificate
1997-06-30
2001-03-20
Copenheaver, Blaine (Department: 1771)
Stock material or miscellaneous articles
Web or sheet containing structurally defined element or...
Adhesive outermost layer
C428S350000, C428S3550AC, C435S030000
Reexamination Certificate
active
06203900
ABSTRACT:
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a pressure sensitive adhesive sheet for the detection of microorganisms on the surface of a test object (hereinafter said surface is also referred to as a test surface) or counting said microorganisms, and to a method for the detection of microorganisms using the pressure sensitive adhesive sheet.
BACKGROUND OF THE INVENTION
For observation of microorganisms such as bacteria which are present on a test surface but cannot be visually observed, there has heretofore been applied a culture method wherein a microorganism is allowed to form a colony. According to this method, a solid plate medium using an agar and the like is pressed against the test surface to transfer the microorganisms on the test surface to the agar plate medium; microorganisms are cultured as they are under the optimal environment; and the colonies formed on the agar plate medium are counted visually. Examples of this method include a food stamp method using an agar stamp commercially available from Nissui Pharmaceutical Co., Ltd. and membrane filter method.
According to the membrane filter method which uses a commercially available membrane filter capable of trapping microorganisms, the microorganisms are collected by thoroughly wiping a test surface with saline, phosphate buffer and the like and filtering the pooled aqueous solution through a membrane filter and the like to trap the microorganisms on the membrane filter. The obtained microorganisms are sufficiently brought into contact with a liquid medium to allow formation of colonies on the filter, and the colonies are counted.
In addition, Igaku Kensa (Medical Test), vol. 41, No. 3, p. 352 (1992) discloses, as an example of culture method, a film coat method including preparation of a film coated with a medium, bringing same into contact with a test surface and culturing same to harvest the detection target microorganism.
A method such as food stamp method, nevertheless, is often associated with insufficient collection efficiency of microorganisms, since it has weak adhesive strength available for transferring microorganisms by pressing a medium against a test surface, and is poor in reproducibility because of variable water contents of the agar medium. Being commonly problematic in various culture methods, contamination among microorganisms is inevitable, which in turn results in interaction among microorganisms on the medium and precludes pure culture, whereby ultimate identification analysis cannot be performed. What is more, not every microorganism is capable of culture to form colonies on a medium and the culture method is limited to the detection of viable cells alone, exclusive of microorganisms that cannot grow in an ordinary medium; thus the method is associated with possible omission of detection. A material limitation on the culture method is that culture time of 1-2 days or longer is needed, so that a real-time monitoring of microorganism detection is not possible.
In the membrane filter method, moreover, a liquid test object such as an aqueous solution can pass through a filter as it is, but other non-liquid test objects require accumulation of microorganisms by time-consuming cumbersome steps of sampling with a cotton swab, preparation of solutions for washing, and the like. In addition, this method does not elude the above-mentioned defects of the culture method, since the determination of microorganisms is based on the culture and the number of colonies formed on a filter by culture. The film coat method also suffer from various defects of the culture method.
The technique recently developed to detect ATP (adenosine triphosphate) in the cells of microorganisms is only applicable to the microorganisms dispersed in water, so that the method for collecting the microorganisms has been the difficult part. As such, there has not been available an easy and simple method for detection of microorganisms in the prior art techniques.
SUMMARY OF THE INVENTION
It is therefore an object of the present invention to solve the above-mentioned defects of the conventional methods and enable real-time monitoring of the detection and determination of microorganisms such as bacteria.
That is, the present invention provides the following.
1) A pressure sensitive adhesive sheet for the detection of microorganisms, which comprises a laminate of an adhesive layer mainly composed of a water-soluble polymer and a water-permeable membrane which does not allow passage of the microorganisms.
2) The sheet of 1) above, wherein the water-permeable membrane is supported by a water-permeable support.
3) The sheet of 1) above, wherein the water-soluble polymer is easily dissolved in water and passes through the water-permeable membrane when dissolved in water.
4) The sheet of 1) above, wherein the surface of the water-permeable membrane on the adhesive layer side has a smoothness of not more than 20 &mgr;m.
5) The sheet of 1) or 2) above, wherein the surface of the adhesive layer has a contact angle with water of not more than 90°.
6) A pressure sensitive adhesive sheet for the detection of microorganisms, which comprises an adhesive layer having a contact angle with water of not more than 90° and a support, the adhesive layer being formed on the support.
7) The sheet of 1) or 6) above, which has a thickness of 10-100 &mgr;m and an adhesive strength to a Bakelite board of 30 g/12 mm width to 600 g/12 mm width.
8) The sheet of 1) or 6) above, wherein the adhesive layer comprises a chromogenic substrate.
9) The sheet of 8) above, wherein the chromogenic substrate is one member selected from 3-(3,4-dihydroxyphenyl)alanine, 3,3′,5,5 -tetramethylbenzidine and nitro blue tetrazolium.
10) The sheet of 1) or 6) above, wherein the adhesive layer comprises an enzyme.
11) The sheet of 10) above, wherein the enzyme is a phenoloxidase.
12) The sheet of 1) or 6) above, wherein the adhesive layer comprises a chromogenic substrate and an enzyme.
13) The sheet of 1) or 6) above, wherein the adhesive layer comprises an enzyme-labeled antibody.
14) The sheet of 13) above, wherein the enzyme-labeled antibody is against microorganism surface antigen and labeled with a peroxidase or alkaline phosphatase.
15) The sheet of 1) or 6) above, wherein the adhesive layer comprises an enzyme-labeled antibody and a chromogenic substrate.
16) The sheet of 1) or 6) above, wherein the adhesive layer comprises a chromogenic substance.
17) The sheet of 16) above, wherein the chromogenic substance is a fluorescent dye.
18) A method for detecting a microorganism, which comprises bringing the surface of an adhesive layer of a pressure sensitive adhesive sheet into contact with the surface of a test object, and then bringing the surface of the adhesive layer into contact with water, said adhesive layer or water containing a chromogenic reagent.
19) The method of 18) above, wherein the surface of the adhesive layer of the pressure sensitive adhesive sheet of 1) or 6) above is brought into contact with the surface of a test object, and then the surface of the adhesive layer is brought into contact with an aqueous solution containing an enzyme and a chromogenic substrate.
20) The method of 18) above, wherein the sheet is that of 1) or 6) above, the adhesive layer thereof comprising a chromogenic substrate, and the surface of the adhesive layer is brought into contact with an aqueous solution containing an enzyme, after contact of the surface of said adhesive layer with the surface of the test object.
21) The method of 18) above, wherein the sheet is that of 1) or 6) above, the adhesive layer thereof comprising an enzyme, and the surface of the adhesive layer is brought into contact with an aqueous solution containing a chromogenic substrate, after contact of the surface of said adhesive layer with the surface of the test object.
22) The method of 18) above, wherein the sheet is that of 1) or 6) above, the adhesive layer thereof comprising a chromogenic substrate and an enzyme, and the surface of the adhesive layer is brought into contact with wate
Iwama Akio
Kazuse Yoshitaka
Nasu Masao
Saika Takeshi
Senda Shuji
Copenheaver Blaine
Guarriello John J.
Nitto Denko Corporation
Sughrue Mion Zinn Macpeak & Seas, PLLC
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