Method for effecting heterogeneous immunological analysis

• US 5,175,086 A
• Filed: 11/09/1987
• Issued: 12/29/1992
• Est. Priority Date: 01/24/1983
• Status: Expired due to Term

• Current Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Original Assignee: Olympus Optical Corporation Limited (Olympus Corporation)
• Inventors: Takekawa, Hiroshi, Yamada, Takashi
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Beisner, William H.
• Application Number:
  US07/119,278
• Time in Patent Office: 1,877 Days
• Field of Search: 435/7, 435/288, 435/291, 435/292, 435/293, 435/296, 435/299, 435/300, 435/301, 435/313, 435/315, 435/7.1, 435/7.9, 435/7.92, 435/7.93, 435/7.94, 435/7.95, 435/188 , 435/808, 436/518, 436/527, 436/531, 436/808, 436/809, 436/535, 436/43, 436/45, 436/47, 436/48, 436/49, 422/64, 422/65, 422/67, 422/68, 422/73, 422/100, 422/63, 422/102
• US Class Current: 435/7.92
• CPC Class Codes: G01N 2035/00396 where the fluid is a liquid
• CPC Class Codes: G01N 2035/00564 Handling or washing solid phase elements, e.g. beads
• CPC Class Codes: G01N 2035/0441 for samples
• CPC Class Codes: G01N 2035/0465 Loading or unloading the conveyor
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 35/025 having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: Y10S 435/808 Optical sensing apparatus
• CPC Class Codes: Y10T 436/111666 Utilizing a centrifuge or compartmented rotor
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/114165 with step of insertion or removal from test line

Abstract

Method for generating an automatic chemical analyzer for measuring given substances in samples in accordance with an enzyme-immunoassay including a turntable rotated intermittently at a given pitch and holding a number of reaction tubes arranged equidistantly along a periphery of the turntable to define a circular reaction line; a carrier supply device for supplying carriers into reaction vessels one by one at a given position in the reaction line, the carrier having given antibody or antigen fixed thereto; a sample delivery device for pouring given amounts of samples into reaction vessels at a given position in the reaction line; a washing device for washing reaction vessels and carriers contained therein to effect B-F separation; a color reagent delivery device for pouring given amounts of a color reagent into reaction vessels to form test liquids; a colorimeter for photometering the test liquids; and a carrier discharge device for removing carriers out of reaction vessels. For respective samples, a reaction vessel is passed through the washing device by a plurality of times to perform a plurality of washing operations including the B-F separation.

Claims

1. An automatic analysis method for detecting the presence of and analyzing biological substances in biological samples in an immunological manner, comprising:

(a) transporting a plurality of reaction vessels along an endless reaction line, said reaction vessels being provided with carriers onto with an antibody or anitgen has been fixed, said antibody or antigen for binding said biological substances;

(b) delivering said biological samples one per vessel into said reaction vessels;

(c) delivering labeled reagents containing labeling substances into the reaction vessels to initiate an antigen-antibody reaction with said biological samples;

(d) effecting a bound-free separation by separating biological substance bound by said antigen or antibody fixed with the carriers and free sample and labeled reagents by washing said reaction vessels and said carriers onto which said antibody or antigen had been fixed;

(e) measuring said biological substances in the samples by employing said labeling substances of the labeled reagents;

(f) discharging the carriers out of the reaction line; and

(g) wherein during analysis of a specific one of said samples a reaction vessel transported along said endless reaction line is indexed to pass by a same washing position a plurality of times to effect at least a first washing and a second washing by means of a same washing device, at least one of said first washing and said second washing including said washing in step (d) to effect said bound-free separation.

2. A method according to claim 1, wherein the reaction line is a circular reaction line comprising a turntable, said reaction vessels being equidistantly spaced apart in a first circular array at uniform angular positions about a periphery of said turntable, said turntable being rotated intermittently at a given angular distance increment equal to the spacing between said reaction vessels.

3. A method according to claim 2, wherein step (a) includes suppling the carriers one per vessel into the reaction vessels one by one, and after steps (e) and (f) have been effected, step (g) includes washing the reaction vessels to prepare for a next analysis.

4. A method according to claim 3, wherein during a first revolution of the turntable after one of said carriers has been supplied into a reaction vessel in step (a), step (b) comprises delivering one of said samples into the reaction vessel and step (c) comprises delivering one of said labeled reagents into the reaction vessel to effect a first antigen-antibody reaction, said labeled reagents comprising an enzyme-labelled reagent;

during a second revolution of the turntable, step (d) comprises washing the reaction vessel and the carrier therein to effect a first bound-free separation of biological sample bound by said antigen or antibody and free sample and labeled reagents and thereafter step (c) comprises again delivering one of said labeled reagents into the reaction vessel to start a second antigen-antibody reaction;

during a third revolution of the turntable, step (d) comprises washing the reaction vessel and the carrier therein to effect a second bound-free separation of biological sample bound by said antigen or antibody and free sample and labeled reagents and step (e) comprises then delivering a color reagent including an enzyme substrate into the reaction vessel to form a test liquid; and

during a fourth revolution of the turntable, step (e) comprises colorimetering the test liquid, step (f) comprises discharging the carrier from the reaction vessel and step (g) comprises washing the reaction vessel.

5. A method according to claim 4, wherein step (g) comprises washing the reaction vessel and carrier after supply of the carrier into the reaction vessel, but prior to the delivery of the sample into the reaction vessel.

6. A method according to claim 5, wherein prior to the supply of the carrier into the reaction vessel, a buffer solution is poured into the reaction vessel.

7. A method according to claim 4, wherein the supply and discharge of carrier, delivery of sample and reagent, and colorimetry are effected once every time the turntable is rotated by three said angular distance increments.

8. A method according to claim 7, wherein step (g) comprises carrying out a washing every time the turntable is rotated by one said angular distance increment.

9. A method according to claim 7, wherein the washing is effected at any one of three different washing positions every time the turntable is rotated by three distance increments by a washing device having said three said angular different washing positions.

10. A method according to claim 4, wherein on the turntable there are arranged second and third arrays of reaction vessels concentric with said first array and the supply and discharge of carrier, delivery of sample and reagents and measuring of said biological samples are effected for one reaction vessel every time the turntable is rotated by one said angular distance increment, while during both said second revolution and said third revolution, step (d) comprises simultaneously washing three reaction vessels respectively positioned in the first, second and third arrays.

11. A method according to claim 3, wherein during a first revolution of the turntable, step (a) includes supplying one of said carriers into a reaction vessel and then steps (b) and (c) comprise delivering sample and said labeled reagent into the reaction vessel to start an antigen-antibody reaction;

during a second revolution of the turntable, step (d) comprises washing of the reaction vessel and the carrier therein to effect a bound-free separation and step (e) includes (i) delivering a color reagent for selectively reacting with said labeled reagent into the reaction vessel to form a test liquid; and

during a third revolution of the turntable, step (e) comprises colorimetering the test liquid, performing step (f) and thereafter step (g) comprises washing the reaction vessel.

12. A method according to claim 11, wherein step (g) comprises washing the reaction vessel and carrier after supply of carrier, but prior to the delivery of sample and reagent.

13. A method according to claim 11, wherein step (a) comprises delivery of the carrier at a position on said reaction line disposed immediately after a position at which the discharge of carrier is performed in step (f), but prior to the washing position of step (g).

14. A method according to claim 1, wherein the reaction vessels are transported along the endless reaction line constituted by an endless belt which is intermittently rotated at a given distance increment in a vertical plane, said distance increment corresponding to a distance between adjacent reaction vessels disposed on said reaction line.

15. A method according to claim 1, wherein step (c) comprises delivering a plurality of liquids including said labeled reagent and buffer solution into the reaction vessels by means of a single delivery device.

16. A method according to claim 15, wherein step (c) comprises selectively delivering buffer solution, enzyme-labeled reagent and color reagent into the reaction vessels by said single delivery device.

17. A method according to claim 16, wherein the delivery device is washed every time a liquid different from a liquid which has been just delivered is to be delivered.

18. An automatic analysis method for detecting the presence of and analyzing biological substances in biological samples in an immunological manner, comprising:

(a) transporting a plurality of reaction vessels along an endless reaction line, said reaction vessels comprising cuvettes having an antibody or antigen fixed thereto, said antibody or antigen for binding said biological substances;

(b) delivering said biological samples one per vessel into said reaction vessels;

(c) delivering labeled reagents containing labeling substances into the reaction vessels to initiate an antigen-antibody reaction with said biological samples;

(d) effecting a bound-free separation by separating biological substance bound by said antigen or antibody fixed with the reaction vessels and free sample and labeled reagents by washing said reaction vessels and;

(e) measuring said biological substances in the samples by employing said labeling substances of the labeled reagents; and

(f) wherein during analysis of a specific one of said samples a reaction vessel transported along said endless reaction line is indexed to pass by a same washing position a plurality of times to effect at least a first washing and a second washing by means of a same washing device, at least one of said first washing and said second washing including said washing in step (d) to effect said bound-free separation, wherein the reaction line is a circular reaction line comprising a turntable, said reaction vessels being equidistantly spaced apart in a first circular array at uniform angular positions about a periphery of said turntable, said turntable being rotated intermittently at a given angular distance increment equal to the spacing between said reaction vessels, and wherein step (a) includes successively supplying the reaction vessels in the form of cuvettes having the antibody or antigen fixed thereto to cuvette holders provided along said periphery of the turntable, and after the measuring of said biological samples in step (e), the cuvette is discharged from the turntable.

19. A method according to claim 18, wherein during a first revolution of the turntable after the supply of a cuvette into the turntable, step (b) includes delivering a sample into the cuvette and step (c) comprises delivering one of said labeled reagents into the cuvette to effect a first antigen-antibody reaction;

during a second revolution of the turntable, step (d) comprises washing the cuvette to effect a first bound-free separation and step (c) comprises then delivering one of said labeled reagents into the cuvette to start a second antigen-antibody reaction; and

during a third revolution of the turntable, step (d) comprises washing the cuvette to effect a second bound-free separation and step (e) comprises delivering a color reagent including an enzyme substrate into the cuvette to form a test liquid; and

during a fourth revolution of the turntable;

step (e) comprises colorimetering the test liquid and step (f) comprises discharging the cuvette from the turntable.

20. A method according to claim 19, wherein step (f) comprises washing the cuvette after supply of the cuvette into the turntable, but prior to the delivery of the sample into the cuvette.

21. A method according to claim 19, wherein the supply and discharge of cuvette, delivery of sample and reagent, and measuring of said biological samples are effected once every time the turntable is rotated by three said angular distance increments.

22. A method according to claim 21, wherein step (f) comprises carrying out a washing every time the turntable is rotated by one of said angular distance increments.

23. A method according to claim 21, wherein step (f) comprises carrying out a washing every time the turntable is rotated by three of said angular distance increments by a washing device having three different washing positions.

24. A method according to claim 18, wherein during a first revolution of the turntable, step (a) comprises supplying a cuvette onto the turntable and steps (b) and (c) respectively comprise delivering sample and said labeled reagent into the cuvette to initiate an antigen-antibody reaction;

during a second revolution of the turntable, step (d) comprises washing the cuvette to effect a bound-free separation and step (e) includes delivering a color reagent for selectively reacting with said labeled reagent into the cuvette to form a test liquid; and

during a third revolution of the turntable, step (e) comprises colorimetering the test liquid and step (f) comprises discharging the cuvette from the turntable.

25. A method according to claim 24, wherein step (f) comprises washing the cuvette after supply of the cuvette, but prior to the delivery of sample and reagent.

26. A method according to claim 24, wherein step (a) comprises delivery of the cuvette at a position on said reaction line disposed immediately after the position at which the discharge of cuvette is performed in step (f) but prior to the washing position of step (f).

27. An automatic analysis method for detecting the presence of and analyzing biological substances in biological samples in an immunological manner, comprising:

(a) transporting a plurality of reaction vessels along an endless reaction line, said reaction vessels being provided with an antibody or antigen fixed onto at least a part of the inner walls of said reaction vessels, said antibody or antigen for binding said biological substances;

(b) delivering said biological samples one per vessel into said reaction vessels;

(c) delivering labeled reagents containing labeling substances into the reaction vessels to initiate an antigen-antibody reaction with said biological samples;

(d) effecting a bound-free separation by separating biological substance bound by said antigen or antibody fixed with said inner walls of the reaction vessels and free sample and labeled reagents by washing said reaction vessels and said antibody or antigen fixed onto at least a part of the inner walls of the reaction vessels;

(e) measuring said biological substances in the samples by employing said labeling substances of the labeled reagent;

(f) discharging the reaction vessels out of the reaction line; and

(g) wherein during analysis of a specific one of said samples a reaction vessel transported along said endless reaction line is indexed to pass by a same washing position a plurality of times to effect at least a first washing and a second washing by means of a same washing device, at least one of said first washing and said second washing including said washing in step (d) to effect said bound-free separation.