Automatic cycling reaction apparatus and automatic analyzing apparatus using the same

• US 4,981,801 A
• Filed: 05/15/1985
• Issued: 01/01/1991
• Est. Priority Date: 05/15/1984
• Status: Expired due to Fees

• Current Assignee: University of Tokyo
• Original Assignee: University of Tokyo
• Inventors: Kato, Takahiko, Suzuki, Yoshiyuki
• Primary Examiner(s): Kamen, Noah P.
• Application Number:
  US06/734,215
• Time in Patent Office: 2,057 Days
• Field of Search: 435/289, 435/290, 435/291, 422/52, 422/64, 422/67
• US Class Current: 435/286.1
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: G01N 2035/00366 Several different temperatures used
• CPC Class Codes: G01N 2035/00396 where the fluid is a liquid
• CPC Class Codes: G01N 2035/00435 Refrigerated reagent storage
• CPC Class Codes: G01N 2035/00445 Other cooling arrangements
• CPC Class Codes: G01N 2035/042 moved independently, e.g. by fork manipulator
• CPC Class Codes: G01N 2035/106 by sucking and blowing
• CPC Class Codes: G01N 35/02 using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations G01N35/0098 and G01N35/0099 take precedence
• CPC Class Codes: G01N 35/025 having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: G01N 35/1004 Cleaning sample transfer devices

Abstract

An apparatus for carrying out an enzymatic cycling reaction including a turntable arranged in a reaction tank, a number of reaction vessels being arranged in the turntable along its periphery, a device for circulating an antifreeze liquid through the reaction tank, a heater for heating the antifreeze liquid, a refrigerator for cooling the antifreeze liquid and a switching valve for selectively passing the antifreeze liquid through the heater or refrigerator. Until all the samples and cycling mixture are delivered into the reaction vessels, the reaction tank is held at a very low temperature at which no enzymatic cycling reaction proceeds. Then the antifreeze liquid is heated to an enzymatic cycling reaction temperature so as to perform the enzymatic cycling reaction simultaneously for liquids contained in all the reaction vessels for a desired period. Then, the antifreeze liquid is heated to a high temperature so as to stop simultaneously the cycling reaction in all the reaction vessels.

Claims

1. An automatic enzymatic cycling reaction apparatus comprising;

first means including a turntable for supporting a plurality of reaction vessels along its periphery, each of said vessels containing given amounts of a sample and a cycling mixture including enzymes; and

second means for holding liquids contained in the reaction vessels at a low temperature at which the enzymatic cycling reaction does not substantially occur, bringing simultaneously liquids contained in all the reaction vessels at a given cycling reaction temperature, keeping the liquids at said temperature for a given period, holding all the liquids simultaneously at a first temperature at which a cycling reaction is stopped due to loss of activity of enzymes, and then keeping all the liquids simultaneously at a second temperature lower than the first temperature, said second means comprising a reaction tank in which said turntable is arranged, means for circulating a thermostatic liquid through said tank and means for controlling the temperature of said thermostatic liquid.

2. An apparatus according to claim 1, wherein said device for controlling the temperature of the thermostatic liquid comprises a heater and a refrigerator and a switching valve for selectively passing the thermostatic liquid through the heater or refrigerator.

3. An apparatus according to claim 2, wherein said second means further comprises a lid removably placed on the reaction tank.

4. An automatic analyzer using an enzymatic cycling reaction comprising;

first means including a turntable for supporting a number of reaction vessels along its periphery each of said vessels containing given amounts of a sample and a cycling mixture including enzymes;

second means for keeping liquids contained in the reaction vessels at a low temperature at which the enzymatic cycling reaction does not substantially occur bringing simultaneously liquids contained in all the reaction vessels at a first temperature at which an enzymatic cycling reaction proceeds, keeping the liquids at said temperature for a given period, holding all the liquids simultaneously at a second temperature at which the enzymatic cycling reaction is stopped due to a loss of activity of the enzymes, and keeping all the liquids simultaneously at a third temperature lower than the second temperature and at which temperature an indicator reaction proceeds;

third means for delivering a given amount of an indicator reagent into respective reaction vessels after the enzymatic cycling reaction of the liquids in the reaction vessels has been stopped; and

fourth means for measuring fluorescent light emitted from substances produced by the indicator reaction, said second means comprising a reaction tank in which said turntable is arranged, means for circulating a thermostatic liquid through said reaction tank, and means for controlling the temperature of said thermostatic liquid.

5. An analyzer according to claim 4, wherein said temperature controlling device comprises a heater, a refrigerator and a switching valve for selectively passing the thermostatic liquid through the heater or refrigerator.

6. An analyzer, according to claim 5, wherein said second means further comprises a lid removably placed on the reaction tank.