Method For Nucleic Acid Detection, Primer For Nucleic Acid Detection, And Kit For Nucleic Acid Detection
AssigneeEiken Kagaku Kabushiki Kaisha
InventorSatoru Michiyuki
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Abstract
[Problem] An objective of the present invention is to develop a new type of method for nucleic acid detection that can be suitably used for detection of short nucleic acids. [Solution] The present invention provides a method for nucleic acid detection including i) hybridizing a detection target nucleic acid 1 and a primer for nucleic acid detection 3 to form a DNA-RNA complex, ii) removing a first polynucleotide segment 301 from the primer for nucleic acid detection 3 by degrading at least a portion of the DNA-RNA complex with a nuclease 4, and iii) hybridizing the primer for nucleic acid detection 3 to a template nucleic acid 2 to perform a reaction for amplifying the template nucleic acid 2 by a polymerase 5.
Claims22
A method for nucleic acid detection for detecting a detection target nucleic acid, using at least
- A) a template nucleic acid that is amplified and serves as a detection index,
- B) a primer for nucleic acid detection including a first polynucleotide segment having a base sequence complementary to at least a portion of the detection target nucleic acid and a second polynucleotide segment having a base sequence complementary to a portion of the template nucleic acid, wherein
- when the detection target nucleic acid is RNA, at least the 5′ end of the first polynucleotide segment is DNA, and when the detection target nucleic acid is DNA, at least the 5′ end of the first polynucleotide segment is RNA, and
- the second polynucleotide segment is linked to the 5′ side of the first polynucleotide segment,
- C) a nuclease that degrades a DNA-RNA complex, and
- D) a polymerase,
- the method comprising:
- i) contacting a test sample containing the detection target nucleic acid with the primer for nucleic acid detection to hybridize the detection target nucleic acid and the primer for nucleic acid detection to form a DNA-RNA complex,
- ii) removing the first polynucleotide segment from the primer for nucleic acid detection by degrading at least a portion of the DNA-RNA complex with the nuclease, and
- iii) hybridizing the primer for nucleic acid detection to the template nucleic acid and amplifying the template nucleic acid with the polymerase.
A primer for nucleic acid detection for detecting a detection target nucleic acid, comprising:
- a first polynucleotide segment having a base sequence complementary to at least a portion of the detection target nucleic acid, and
- a second polynucleotide segment having a base sequence complementary to a portion of the template nucleic acid that is amplified and serves as a detection index, wherein
- when the detection target nucleic acid is RNA, at least the 5′ end of the first polynucleotide segment is DNA, and when the detection target nucleic acid is DNA, at least the 5′ end of the first polynucleotide segment is RNA, and
- the second polynucleotide segment is linked to the 5′ side of the first polynucleotide segment.
A kit for nucleic acid detection for detecting a detection target nucleic acid, comprising:
- A) a template nucleic acid that is amplified and serves as a detection index,
- B) a primer for nucleic acid detection including a first polynucleotide segment having a base sequence complementary to at least a portion of the detection target nucleic acid and a second polynucleotide segment having a base sequence complementary to a portion of the template nucleic acid, wherein
- when the detection target nucleic acid is RNA, at least the 5′ end of the first polynucleotide segment is DNA, and when the detection target nucleic acid is DNA, at least the 5′ end of the first polynucleotide segment is RNA, and the second polynucleotide segment is linked to the 5′ side of the first polynucleotide segment, and
- C) a nuclease that degrades a DNA-RNA complex.
Dependencies are inferred from the claim text.