Method and reaction kit for agglutination detection

• US 5,188,968 A
• Filed: 03/02/1992
• Issued: 02/23/1993
• Est. Priority Date: 12/28/1989
• Status: Expired due to Term

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Olympus Optical Company Ltd. (Olympus Corporation)
• Inventors: Kano, Tokio, Niimura, Toshinobu, Yonekawa, Hiroyuki
• Primary Examiner(s): Hill, Jr., Robert J.
• Application Number:
  US07/841,769
• Time in Patent Office: 358 Days
• Field of Search: 422/58, 422/61, 422/73, 422/102, 436/165, 436/501, 436/520, 436/533, 436/539, 436/541, 424/11
• US Class Current: 436/501
• CPC Class Codes: B01L 2300/0822 Slides
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 9/52 Supports specially adapted for flat sample carriers, e.g. for plates, slides, chips
• CPC Class Codes: G01N 33/5304 Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

A reaction kit is provided with a reaction zone of suitable cross-sectional area to aspirate a sample by capillarity and a transparent plate with a flat surface in at least part of the reaction zone, and a supporting base to incline the reaction zone at a specified angle. The reaction zone is normally a space formed by two opposite transparent plates and spacers inserted between these two transparent plates. A supporting base may be molded separately, the transparent plates and spacers being set on the base at the time of measurement, or alternatively the base may be molded integrally with at least one of the plates defining the reaction zone. Further, substances with specificity such as antigens or antibodies may be coated on a surface in the reaction zone. The occurrence or non-occurrence of an agglutination reaction may be determined by introducing a particle suspension containing particles with specific bonding properties and a substance with specific bonding properties into the reaction zone by means of capillarity, leaving the apparatus for a predetermined time to carry out the reaction, and thereafter observing the particle distribution pattern.

Claims

1. A reaction kit for observation and detection of a particle pattern due to an agglutination reaction, comprising:

reaction zone defining means for receiving a sample, said reaction zone defining means comprising;

two plates arranged opposed to each other, at least one of said plates being transparent for visual detection therethrough of a particle pattern formed in said reaction zone; and

spacers inserted between said two plates so as to maintain said two plates in spaced apart relation to each other, said reaction zone being defined in a space between said two plates and between said spacers; and

a supporting base for supporting said reaction zone defining means, and including means for inclining said reaction zone defining means at a specified angle to thereby incline said reaction zone, such that one of said spacers forms a bottom portion of the inclined reaction zone;

a part of the space defined between said two plates and between said spacers serving as an opening through which a sample is introduced into said reaction zone, and wherein said opening is located in a position different from said spacer forming the bottom portion of the inclined reaction zone;

wherein a sample is aspirated into said reaction zone when the sample is applied to said reaction zone defining means.

2. The reaction kit of claim 1, wherein said two plates are both transparent.

3. The reaction kit of claim 1, wherein said reaction zone defining means and said supporting base are integrally formed as one unit.

4. The reaction kit of claim 1, wherein said reaction zone defining means includes a bottom surface facing said reaction zone, and further comprises a substance with specific bonding affinity to a sample, coated on at least said bottom surface of said reaction zone defining means.

5. The reaction kit of claim 1, wherein said spacers comprises two spaced apart spacers inserted between said two plates, said two spacers being spaced apart in a direction parallel to said two plates, and the reaction zone being defined in the space between said two spacers.

6. The reaction kit of claim 5, wherein said spacers are substantially flat plates.

7. The reaction kit of claim 5, wherein said reaction zone defining means defines said reaction zone having a substantially rectangular cross-section.

8. The reaction kit of claim 5, wherein said reaction zone defining means said reaction zone having a substantially rectangular cross-section.

9. The reaction kit of claim 1, wherein said spacers include surface means for defining an inlet port in flow communication with said reaction zone, said inlet port having a greater cross-sectional area than said reaction zone.

10. The reaction kit of claim 9, wherein said spacers comprise two spaced apart spacer plates inserted between said two plates, said two spacers being spaced apart in a direction parallel to said two plates, and the reaction zone being defined in the space between said two spacers, at least one of said spacer plates having an angled surface to define an inlet port in flow communication with said reaction zone, said inlet port having a greater cross-sectional area than said reaction zone.

11. The reaction kit of claim 1, wherein said reaction zone defining means further comprises:

means defining a trough adjacent said reaction zone.

12. The reaction kit of claim 11, wherein said reaction zone;

and

means defining a trough adjacent the opposite openings of said reaction zone.

13. A method of determining a degree of particle agglutination by an agglutination reaction having the reaction kit according to claim 1, the method comprising:

preparing a particle suspension containing particles with specific bonding properties and a substance with specific bonding affinity to the particles;

introducing the particle suspension into a reaction zone of said reaction kit according to claim 1;

setting said reaction zone defining means on said supporting base, and leaving said reaction zone defining means, with the particle suspension therein, on said supporting base for a predetermined time; and

observing the formation and degree of spread of a particle distribution pattern formed in a lower part of said reaction zone defined by said reaction zone defining means.