Method for detecting DNA with probe PNA
AssigneeIsao Karube
InventorsIsao Karube, Shinya Sawata, Ryohei Nagata
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Abstract
An object of the present invention is to provide a method for detecting a target nucleotide sequence using the hybridization method, which has a remarkably improved sensitivity of detection. The method comprises the steps of hybridizing a target nucleotide sequence with a PNA that is complementary to the whole or a part of the target nucleotide sequence and measuring the degree of hybridization at the presence of a denaturing agent.
Claims20
A method for detecting a target double stranded DNA, which comprises the steps of:
- hybridizing the target double stranded DNA with a single stranded PNA (peptide nucleic acid) which is complementary to the whole or a part of the target DNA;
- and
- measuring the degree of hybridization at the presence of a denaturing agent.
A method for detecting Escherichia coli O-157. which comprises the steps of:
- amplifying a genome DNA of Escherichia coli O-157 by PCR to obtain a double stranded DNA;
- hybridizing the double stranded DNA with a single stranded PNA which has the same sequence as at least 15 consecutive nucleotides of the nucleotide sequence of SEQ ID NO:
- 1;
- and
- measuring the degree of hybridization at the presence of a denaturing agent.
A method for detecting Escherichia coli O-157, which comprises the steps of:
- amplifying a genome DNA of Escherichia coli O-157 by PCR to obtain a double stranded DNA by using a sense primer selected from SEQ ID NOS. 4, 5 7, 8, and 9 and an antisense primer of SEQ ID NO. 6;
- hybridizing the double stranded DNA with a single stranded PNA selected from the group consisting of the sequences of SEQ ID NOS:2, 16, and 17 and a complementary sequence thereof;
- and
- measuring the degree of hybridization by using a surface plasmon resonance biosensor at the presence of a denaturing agent at a temperature not exceeding 40° C.
An apparatus for detecting Escherichia coli O-157, comprising
- a surface plasmon resonance biosensor;
- a measuring chip for the surface plasmon resonance biosensor;
- and
- a single Stranded PNA selected from the group consisting of the sequences of SEQ ID NOS:2, 16, and 17 and a complementary sequence thereof, which is immobilised on a surface of the measuring chip.
Dependencies are inferred from the claim text.