Dual chamber disposable reaction vessel for amplification reactions, reaction processing station therefor, and methods of use
First Claim
1. A dual chamber reaction vessel for a nucleic acid amplification reaction, the vessel comprising:
- a first chamber for receiving a fluid sample, said first chamber loaded with an amplification reagent(s);
a second chamber physically isolated from said first chamber;
an enzyme(s) for said reaction either in said second chamber or in fluid communication with said second chamber; and
a fluid channel connecting said first chamber to said second chamber.
5 Assignments
0 Petitions
Accused Products
Abstract
A reaction vessel for a nucleic acid amplification reaction has a first chamber containing an amplification reagent mix, a second chamber containing an amplification enzyme, and a fluid channel or chamber connecting the first and second chambers together. A fluid sample is introduced into the first chamber. After a denaturation and primer annealing process has occurred in the first chamber, the fluid channel is opened to allow the solution of the reagent and fluid sample to flow into the second chamber. The second chamber is maintained at an optimal temperature for the amplification reaction.
A station is described for processing test strips incorporating the reaction vessels. The station includes temperature and vacuum control subsystems to maintain proper temperatures in the reaction vessel and effectuate the transfer of the fluid from one chamber to the other in an automated fashion without human intervention.
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Citations
25 Claims
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1. A dual chamber reaction vessel for a nucleic acid amplification reaction, the vessel comprising:
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a first chamber for receiving a fluid sample, said first chamber loaded with an amplification reagent(s); a second chamber physically isolated from said first chamber; an enzyme(s) for said reaction either in said second chamber or in fluid communication with said second chamber; and a fluid channel connecting said first chamber to said second chamber. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 11, 13, 23)
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9. A reaction vessel for a nucleic acid amplification reaction, comprising:
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an amplification well; a channel separated from said amplification well by a heat and moisture barrier; a carrier in said channel carrying an amplification enzyme; wherein said carrier is moveable within said channel such that said carrier may be moved through said heat and moisture barrier to deliver said enzyme to said amplification well.
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10. A two-piece disposable reaction vessel, comprising:
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a first piece comprising a first chamber containing an amplification reagent; a second piece comprising a second chamber containing an amplification enzyme; said first and second pieces constructed so as permit said first and second pieces to be securely joined into an dual chamber reaction vessel such that a fluid sample in said first piece may pass into said second chamber.
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12. In a test strip for a nucleic acid amplification reaction comprising a body portion defining a plurality of wells arranged in a row and having a first end and a second end, said second end having a cuvette for conducting an optical analysis of a sample, the improvement comprising:
providing an aperture in said body portion adjacent to said first end of said test strip for receiving a disposable amplification reaction vessel.
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14. A method for conducting a nucleic acid amplification reaction, comprising the steps of:
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providing a dual chamber reaction vessel comprising first and second chambers capable of being placed in fluid communication with each other, said dual chamber reaction vessel loaded with an amplification reagent(s) into said first chamber and an amplification enzyme(s) in said second chamber; sealing said first and second chambers with said amplification reagent(s) and amplification enzyme(s) from the environment with a sealing means after loading said amplification reagent(s) and amplification enzyme(s) in said dual chamber reaction vessel; and, at the time of use of said dual chamber reaction vessel; opening said sealing means with an implement and loading a sample into said first chamber; heating said first chamber and said sample to a first relatively high temperature while maintaining said second chamber at a second relatively low temperature; cooling said first chamber and a solution of said sample and said amplification reagent(s) from said first relatively high temperature; thereafter passing said solution from said first chamber to said second chamber; and amplifying said solution in said second chamber by said amplification enzyme(s). - View Dependent Claims (15)
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16. A method for conducting a nucleic acid amplification reaction, comprising the steps of:
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providing a reaction vessel comprising an amplification chamber and a second chamber separated from said amplification chamber by a heat and moisture barrier; loading an amplification reagent(s) into said amplification chamber and an amplification enzyme(s) in said second chamber; loading a sample into said amplification chamber; heating said first chamber and said sample to a first relatively high temperature greater than or equal to 65 degrees C. while maintaining said enzyme in said second chamber at a relatively cooler second temperature; cooling said first chamber and a solution of said sample and said amplification reagent(s); and thereafter introducing said amplification enzyme(s) into said amplification chamber by forcing said enzyme(s) through said heat and moisture barrier into said amplification chamber; and amplifying said solution in said amplification chamber.
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17. A nucleic acid amplification station for a test strip, said test strip comprising a dual chamber reaction vessel comprising a first chamber and a second chamber, the station comprising:
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a tray for at least one test strip, said tray comprising a first portion and a second portion positioned adjacent to first and second chambers of said dual chamber reaction vessel, respectively; a temperature control subsystem for said tray maintaining said first and second portions of said tray at first and second different amplification reaction temperatures so as to maintain said first and second chambers at said first and second amplification reaction temperatures, respectively; a fluid conduit opening mechanism for opening a fluid conduit in said dual chamber reaction vessel to thereby establish fluid communication between said first chamber and said second chamber; and a vacuum subsystem including a vacuum probe, said test strip and vacuum probe reciprocable relative to each other, said vacuum probe cooperating with said reaction wells in said test strip for transferring a fluid sample from said first chamber to said second chamber via said fluid conduit. - View Dependent Claims (18, 19, 20, 21, 22)
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24. A nucleic acid amplification station for a test strip, said test strip comprising a dual chamber reaction vessel comprising a first chamber and a second chamber, the station comprising:
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a tray for at least one test strip, said tray comprising a first portion and a second portion positioned adjacent to first and second chambers of said dual chamber reaction vessel, respectively; a temperature control subsystem for said tray maintaining said first and second portions of said tray at first and second different amplification reaction temperatures so as to maintain said first and second chambers at said first and second amplification reaction temperatures, respectively, during processing of said test strip; and a fluid conduit opening mechanism for opening a fluid conduit in said dual chamber reaction vessel to thereby establish fluid communication between said first chamber and said second chamber.
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25. A method for conducting a nucleic acid amplification reaction, comprising the steps of:
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providing a dual chamber reaction vessel comprising first and second chambers capable of being placed in fluid communication with each other, said dual chamber reaction vessel loaded with an amplification reagent(s) into said first chamber and an amplification enzyme(s) in said second chamber; sealing said first and second chambers with said amplification reagent(s) and amplification enzyme(s) from the environment with a sealing means after loading said amplification reagent(s) and amplification enzyme(s) in said dual chamber reaction vessel; and, at the time of use of said dual chamber reaction vessel; opening said membrane with an implement and loading a sample into said first chamber; heating said first chamber and said sample to a first relatively high temperature while maintaining said second chamber at a second relatively low temperature; cooling said first chamber and a solution of said sample and said amplification reagent(s) from said first relatively high temperature; thereafter introducing the contents of said second chamber into said first chamber; and amplifying said sample in first chamber with said amplification enzyme(s).
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Specification