Device for rapid detection and identification of single microorganisms without preliminary growth

• US 20050221403A1
• Filed: 04/20/2005
• Published: 10/06/2005
• Est. Priority Date: 05/09/2002
• Status: Active Grant

• Current Assignee: NanoLogix, Inc.
• Original Assignee: NanoLogix, Inc.
• Inventors: Gazenko, Sergey
• Granted Patent: US 8,663,909 B2
• Time in Patent Office: Days
• US Class Current: 435/7.320
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence

Abstract

This invention describes a device consisting of a micro channel plate, filter, and porous holder for filter, which is substituted by a pure agar block during method performance, and supportive structural elements. The device is intended for rapid detection and/or identification of microorganisms. Microorganisms are trapped by filtration in long (diameter/length=1/10-1/100), cylindrical, parallel, micro-channels that are open from both sides and attached to a filter from one side. A micro channel plate houses a multiplicity of micro channels (possible diameter of each channel=1-30 μm, length 100-1000 μm, and number on centimeter²−100,000-1,000,000). The micro channel plate with cells trapped on the surface of the filter is attached to an agar block impregnated by artificial substrate(s) so that the molecules of the artificial substrates will fill all micro channels. Trapped cells produce colored or fluorescent molecules from artificial substrates. These molecules are collected in the very small volume of a micro channel. The extremely small volume of a micro channel (1/25 million part of milliliter) allows it to collect a detectable concentration of color or fluorescent substances in a very short time (several minutes). Even one cell from a filtrated sample can be detected by the enzyme—artificial substrate method and/or identified by enzyme immunoassay.

Claims

1. A device for detection and identification of microorganisms without preliminary growth by depositing microorganmsms in micro-channels of a small size (diameter:

1-30 μ

m;

length;

100-1000 μ

m;

volume;

1000-300,000 μ

k³) and providing enzyme or enzyme immunoassay with rapid creation of a detectable concentration of indicator substances which device comprises;

a micro-channel plate attached to the filtrating material;

liquids and air transparent holder pressed to filter and frame to support all structure;

this holder changes to an agarose block filled by artificial substrate(s), which fill all micro-channels of the micro-channel plate by capillary forces and, initiates colored or fluorescent reactions in micro-channels containing microorganisms

2. Device according to claim 1 in which the micro-channel plate is colorless and the filtrating material is colorless or white in order to pass light through the device under a light microscope and make colored micro-channels more visible

3. Device according to claim 1 in which the micro-channel plate and filter are black in order to limit background fluorescence when fluorescent molecules are produced in micro-channels containing microorganisms and fluorescent microscope is used for detection of fluorescence in the micro-channels

4. Device according to claim 1 where micro-channel plate and filter are black or colorless or white but transparent holder for filter is substituted by a magnet for depositing of magnetic particles mated by antibodies to cells, viruses, or biomolecules and antigens (cells, viruses, or biomolecules) attached to named antibodies through antibody-antigen interaction

5. Device according to claim 1 where the micro-channel plate used for performing color or fluorescent reactions is a micro-channel glass plate used in image intensifiers with angle between channels to the surface of plate of 90°

and the volumes of micro-channels in a range of 1000-300,000 μ

m³

6. Device according to claim 1 where the matters for storage of artificial substrates and their transfer to micro-channels could be gels (agarose, gelatin, polyacrylamide, silica gels), paper and porous material capable of keeping liquid with dissolved molecules inside and transfer to micro-channels.