Thermal reaction device and method for using the same

A method for carrying out nucleic acid amplification reactions to detect the quantity, presence, or absence of a plurality of target polynucleotide sequences in a sample, the method comprising:

• (a) providing a microfluidic device comprising
• (i) an inlet,
• (ii) a first flow channel in fluid communication with the inlet, and
• (ii) a plurality of second channels in fluid communication with the first flow channel, wherein each second channel comprises an entrance and a dead end,
• wherein amplification primers, and optionally other amplification reagents, are non-covalently deposited at reaction sites at the dead ends of each second channel, and amplification primers deposited in different second channels amplify different target polynucleotide sequences,
• (b) introducing a sample solution comprising the sample through the inlet such that the sample solution flows through the first flow channel and into the plurality of second channels, wherein the sample comprises a plurality of polynucleotides, and wherein introduction of the sample solution into the second channels causes the deposited amplification primers to be dissolved into the sample solution, and
• (c) isolating the reaction sites from the first flow channel and from each other,
• (d) conducting polynucleotide amplification reactions in the reaction sites, and
• (e) detecting the quantity, presence or absence of polynucleotide amplification products in the reaction sites to determine the quantity, presence, or absence of said target polynucleotide sequences in the sample.

A method for carrying out nucleic acid amplification reactions to detect the presence or absence of a plurality of different target viral or bacterial polynucleotide sequences in a biological sample obtained from a human patient, the method comprising:

• (a) providing a microfluidic device comprising
• (i) an inlet,
• (ii) a first flow channel in fluid communication with the inlet, and
• (iii) a plurality of second channels in fluid communication with the first flow channel, wherein each second channel comprises an entrance and a dead end,
• wherein amplification primers, and optionally other amplification reagents, are non-covalently deposited at a reaction site at the dead-ends of each second channel, and amplification primers deposited in different second channels amplify different target polynucleotide sequences;
• and
• wherein the different target polynucleotide sequences are viral or bacterial sequences,
• (b) introducing a sample solution comprising the sample through the inlet such that the sample solution flows through the first flow channel and into the plurality of second channels, wherein the sample comprises a plurality of polynucleotides, and wherein introduction of the sample solution into the second channels causes the deposited amplification primers to be dissolved into the sample solution,
• (c) isolating the reaction sites from the first flow channel and from each other,
• (d) conducting polynucleotide amplification reactions in the reaction sites, and
• (e) detecting the presence or absence of a polynucleotide amplification products in the reaction sites to determine the presence or absence of a viral or bacterial target polynucleotide sequence in the biological sample obtained from the human patient.