Nucleic hybridization assay method and device using a cleavage technique responsive to the complementary double strand or the single strand of nucleic acids or oligonucleotides
First Claim
1. A cleavable signal element comprising:
- a restriction probe of a single strand having a particular sequence cleavable by a restriction enzyme specific to a double strand of a particular sequence; and
a capture probe of a single strand having a complementary sequence to a target nucleic acid for diagnosis or assay to form a double strand through hybridization to the target nucleic acid, wherein one end of the restriction probe is attached to a solid support substrate, and the other end of the restriction probe is ligated to the capture probe, thus forming a single-stranded, cleavable capture probe.
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Accused Products
Abstract
A cleavable signal element applicable to quantitative and qualitative assay devices, using a cleavable technique specifically responsive to a complementary double strand or single strand of nucleic acids, and a nucleic acid hybridization assay method and device using the cleavable signal element are provided. Using the cleavable technique responsive to the complementary double strand or single strand of nucleic acids, detection sensitivity to a target nucleic acid can be increased, and diagnosis and detection reliability can be improved twice through in-situ determinations. Through simultaneous single nucleotide polymorphism (SNP) detection and expression profile determination, more accurate diagnosis for many diseases can be achieved. The assay device can be easily modified to be suitable for detection with general laser-based detection systems such as CD-ROM readers. Information read from the assay device is digitized as software and transmitted to and received by doctors and patients through a computer network or wirelessly, which enables construction of remote diagnosis systems.
56 Citations
83 Claims
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1. A cleavable signal element comprising:
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a restriction probe of a single strand having a particular sequence cleavable by a restriction enzyme specific to a double strand of a particular sequence; and
a capture probe of a single strand having a complementary sequence to a target nucleic acid for diagnosis or assay to form a double strand through hybridization to the target nucleic acid, wherein one end of the restriction probe is attached to a solid support substrate, and the other end of the restriction probe is ligated to the capture probe, thus forming a single-stranded, cleavable capture probe. - View Dependent Claims (2, 3, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79)
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- 4. A cleavable signal element comprising a capture probe of a single strand having a complementary sequence to a target nucleic acid for diagnosis or assay to form a double strand through hybridization to the target nucleic acid, wherein one end of the capture probe is attached to a solid support substrate, and the capture probe itself forms a single-stranded, cleavable capture probe which is cleavable by a cleavage enzyme specifically responsive to a double strand or single strand of nucleic acids.
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80. A nucleic acid hybridization assay method comprising:
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(a) injecting a sample into a sample injection port disposed near the center of a disk in a nucleic acid hybridization assay device;
(b) rotating the disk and stopping the rotation of the disk when the sample reaches an outer edge of the disk;
(c) incubating the disk in a stationary state at room temperature for hybridization;
(d) adding a buffer solution as a washing solution while rotating the disk at a high speed, to wash the disk;
(e) adding a DNA polymerization solution containing a mixed solution of four dNTPs and a DNA polymerase and incubating the disk in a stationary state for DNA extension;
(f) adding a solution of a restriction enzyme specifically responsive to a double strand of a particular sequence and incubating the disk in a stationary state, to cleave the double strand;
(g) washing the disk by rotating the disk at a high speed with the addition of a buffer solution or by applying an external electric or magnetic field; and
(h) drying the disk and reading information from the disk using a detector which is programmed to detect a predetermined assay site on which a cleavable signal element is deposited and comprises an optical device, an electrochemical device, or a capacitance and impedance measurement device. - View Dependent Claims (81, 83)
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82. A nucleic acid hybridization assay method comprising:
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(a) injecting a sample into a sample injection port disposed near the center of a disk in a nucleic acid hybridization assay device;
(b) rotating the disk and stopping the rotation of the disk when the sample reaches an outer edge of the disk;
(c) incubating the disk in a stationary state at room temperature for hybridization;
(d) adding a buffer solution as a washing solution while rotating the disk at a high speed, to wash the disk;
(e) adding a solution of a cleavage enzyme specifically responsive to a double strand or single strand of nucleic acids and incubating the disk in a stationary state, to cleave the double strand or single strand;
(f) washing the disk by rotating the disk at a high speed with the addition of a buffer solution or by applying an external electric or magnetic field; and
(g) drying the disk and reading information from the disk using a detector which is programmed to detect a predetermined assay site on which a cleavable signal element is deposited and comprises an optical device, an electrochemical device, or a capacitance and impedance measurement device.
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Specification