US 2016/0257992 A1Published application

Dna detection method and dna detection device

AssigneePanasonic Intellectual Property Management Co., Ltd.

InventorMASAHIKO TSUKUDA

Life of this patent

Estimating the term…

Abstract

Disclosed is a DNA detection device that detects a target DNA by detecting fluorescence output from sample droplets flowing through a flow path in a surface of a sensor chip, and that includes a fluorescence detector that detects fluorescence output from sample droplets flowing through the flow path, and a DNA detector that determines a type of fluorescent probe solution contained in each of the sample droplets based on a duration of the detected fluorescence and determines whether or not the sample droplet contains the target DNA based on whether intensity of the fluorescence is higher or lower than a threshold value.

Claims12

Dependent claims

A DNA detection method comprising:

(a) placing a sensor chip in a DNA detection device, wherein
the DNA detection device comprises:
a PCR processor, a fluorescence detector, and a DNA detector,
the sensor chip comprises:
a first flow path, a second flow path, a third flow path, a fourth flow path, a fifth flow path, a sixth flow path, a seventh flow path, an eighth flow path, and a ninth flow path,
a first end of the first flow path and a first end of the second flow path are connected to a first end of the third flow path,
a second end of the third flow path is connected to a first end of the sixth flow path,
the fourth flow path and the fifth flow path are connected between the first end and the second end of the third flow path,
a second end of the sixth flow path and a first end of the seventh flow path are connected to a first end of the eighth flow path,
a second end of the eighth flow path is connected to the PCR processor, and
the PCR processor is connected to the ninth flow path;
(b) introducing an aqueous DNA solution and an aqueous DNA polymerase solution into the first flow path and the second flow path, respectively, to pass a first aqueous mixture of the aqueous DNA solution and the aqueous DNA polymerase solution through the third flow path, wherein the aqueous DNA solution contains a target single-stranded DNA;
(c) introducing a first aqueous fluorescent probe solution obtained by mixing a first fluorescent probe with a first primer into the fourth flow path at a first flow rate during flowing of the first aqueous mixture through the third flow path to pass a second aqueous mixture of the first aqueous mixture and the first aqueous fluorescent probe solution through the sixth flow path, wherein the first fluorescent probe complementarily binds to a first single-stranded DNA;
(d) introducing an oil material into the seventh flow path at a second flow rate to pass parts of the second aqueous mixture and parts of the oil material through the eighth flow path, wherein the parts of the second aqueous mixture and the parts of the oil material are arranged alternately along the eighth flow path;
(e) introducing a second aqueous fluorescent probe solution obtained by mixing a second fluorescent probe with a second primer into the fifth flow path at a third flow rate during flowing of the first aqueous mixture through the third flow path to flow a third aqueous mixture of the first aqueous mixture and the second aqueous fluorescent probe solution through the sixth flow path, wherein the second fluorescent probe is different from the first fluorescent probe and complementarily binds to a second single-stranded DNA;
(f) introducing the oil material into the seventh flow path at a fourth flow rate to pass parts of the third aqueous mixture and parts of the oil material through the eighth flow path, wherein the parts of the third aqueous mixture and the parts of the oil material are arranged alternately along the eighth flow path;
(g) processing the parts of the second aqueous mixture and the parts of the third aqueous mixture by PCR with the PCR processor and then passing the parts through the ninth flow path;
(h) detecting, with the fluorescence detector, intensity of fluorescence output from each of the parts of the second aqueous mixture and the parts of the third aqueous mixture flowing through the ninth flow path;
and
(i) detecting, with the DNA detector, whether or not the target single-stranded DNA contains at least one selected from the first single-stranded DNA and the second single-stranded DNA based on the intensity of transmitted light, the first flow rate, the second flow rate, the third flow rate, and the fourth flow rate.

17 more elements

A DNA detection device that detects a DNA based on fluorescence output from parts of an aqueous DNA mixture flowing through a flow path in a surface of a sensor chip, wherein

the aqueous DNA mixture is an aqueous solution obtained by mixing a DNA mixture with a first aqueous fluorescent probe solution or a second aqueous fluorescent probe solution,
the DNA mixture is an aqueous solution obtained by mixing a DNA solution containing a target single-stranded DNA with a DNA polymerase,
the first aqueous fluorescent probe solution is an aqueous solution obtained by mixing a fluorescent probe with a primer that complementarily bind to a first single-stranded DNA,
the second aqueous fluorescent probe solution is an aqueous solution obtained by mixing a fluorescent probe with a primer that complementarily bind to a second single-stranded DNA, and
the device comprises:
a fluorescence detector that detects intensity of fluorescence output from parts of the aqueous DNA mixture flowing through the flow path:
and
a DNA detector that determines a type of fluorescent probe solution contained in each of the parts of the aqueous DNA mixture based on a duration of the fluorescence and detects whether or not the target single-stranded DNA contains at least one selected from the first single-stranded DNA and the second single-stranded DNA based on whether intensity of the fluorescence is higher or lower than a threshold value.

5 more elements

Dependencies are inferred from the claim text.

patentgrep

Loading patent…

Abstract

A DNA detection method comprising:

(a) placing a sensor chip in a DNA detection device, wherein
the DNA detection device comprises:
a PCR processor, a fluorescence detector, and a DNA detector,
the sensor chip comprises:
a first flow path, a second flow path, a third flow path, a fourth flow path, a fifth flow path, a sixth flow path, a seventh flow path, an eighth flow path, and a ninth flow path,
a first end of the first flow path and a first end of the second flow path are connected to a first end of the third flow path,
a second end of the third flow path is connected to a first end of the sixth flow path,
the fourth flow path and the fifth flow path are connected between the first end and the second end of the third flow path,
a second end of the sixth flow path and a first end of the seventh flow path are connected to a first end of the eighth flow path,
a second end of the eighth flow path is connected to the PCR processor, and
the PCR processor is connected to the ninth flow path;
(b) introducing an aqueous DNA solution and an aqueous DNA polymerase solution into the first flow path and the second flow path, respectively, to pass a first aqueous mixture of the aqueous DNA solution and the aqueous DNA polymerase solution through the third flow path, wherein the aqueous DNA solution contains a target single-stranded DNA;
(c) introducing a first aqueous fluorescent probe solution obtained by mixing a first fluorescent probe with a first primer into the fourth flow path at a first flow rate during flowing of the first aqueous mixture through the third flow path to pass a second aqueous mixture of the first aqueous mixture and the first aqueous fluorescent probe solution through the sixth flow path, wherein the first fluorescent probe complementarily binds to a first single-stranded DNA;
(d) introducing an oil material into the seventh flow path at a second flow rate to pass parts of the second aqueous mixture and parts of the oil material through the eighth flow path, wherein the parts of the second aqueous mixture and the parts of the oil material are arranged alternately along the eighth flow path;
(e) introducing a second aqueous fluorescent probe solution obtained by mixing a second fluorescent probe with a second primer into the fifth flow path at a third flow rate during flowing of the first aqueous mixture through the third flow path to flow a third aqueous mixture of the first aqueous mixture and the second aqueous fluorescent probe solution through the sixth flow path, wherein the second fluorescent probe is different from the first fluorescent probe and complementarily binds to a second single-stranded DNA;
(f) introducing the oil material into the seventh flow path at a fourth flow rate to pass parts of the third aqueous mixture and parts of the oil material through the eighth flow path, wherein the parts of the third aqueous mixture and the parts of the oil material are arranged alternately along the eighth flow path;
(g) processing the parts of the second aqueous mixture and the parts of the third aqueous mixture by PCR with the PCR processor and then passing the parts through the ninth flow path;
(h) detecting, with the fluorescence detector, intensity of fluorescence output from each of the parts of the second aqueous mixture and the parts of the third aqueous mixture flowing through the ninth flow path;
and
(i) detecting, with the DNA detector, whether or not the target single-stranded DNA contains at least one selected from the first single-stranded DNA and the second single-stranded DNA based on the intensity of transmitted light, the first flow rate, the second flow rate, the third flow rate, and the fourth flow rate.

17 more elements