| 主权项 |
1. A method for detecting first nucleic acid targets and second nucleic acid targets using a microfluidic chip, the method comprising:
(a) installing the microfluidic chip in a detection device, wherein the detection device comprises a PCR processor, a PCR reactor, a fluorescence detector, and a detect circuitry, the microfluidic chip comprises a first flow channel, a second flow channel, a third flow channel, a fourth flow channel, and a fifth flow channel, wherein the fourth flow channel is connected with one end of the first flow channel, one end of the second flow channel, and one end of the third flow channel, the PCR reactor is located between the fourth flow channel and the fifth flow channel, (b) (i) supplying from another end of the first flow channel a first aqueous solution, a first oil, and a second aqueous solution in this order, (ii) supplying from another end of the second flow channel a sample aqueous solution, and (iii) supplying a second oil from another end of the third flow channel, thereby causing a first drop, a second drop, a third drop and a fourth drop to pass through the fourth flow channel in this order, wherein the first drop is made from the first aqueous solution and the sample aqueous solution, the second drop is made from the sample aqueous solution, the third drop is made from the second aqueous solution and the sample aqueous solution, the fourth drop is made from the sample aqueous solution, wherein the first aqueous solution has a first DNA having a complementary sequence to the first nucleic acid target, the first DNA is modified with a first fluorescent dye, and the second aqueous solution has a second DNA having a complementary sequence to the second nucleic acid target, the second DNA is modified with a second fluorescent dye, (c) subjecting the first drop; the second drop; the third drop; and the fourth drop which have passed through the fourth flow channel and reached the PCR reactor, to a PCR process with the PCR processor, and causing the first drop; the second drop; the third drop; and the fourth drop which have been subjected to the PCR process to pass through the fifth flow channel, (d) detecting intensities of fluorescence outputted from the first drop to the fourth drop flowing in the fifth flow channel with the fluorescence detector, (e) acquiring boundaries between the first drop; the second drop; the third drop; and the fourth drop flowing in the fifth flow channel based on the intensities of fluorescence, and (f) acquiring a number of the second drop and the fourth drop having an intensity of fluorescence greater than or equal to a first threshold based on the intensity of fluorescence and the boundaries between the first drop; the second drop; the third drop; and the fourth drop, and detecting whether or not the sample aqueous solution include at least one selected from the group consisting of the first nucleic acid target and the second nucleic acid target based on the number of the second drop and the fourth drop with the detect circuitry. |
