Stanniocalcin (STC1) mRNA assay method
AssigneeTosoh Corporation
InventorsJuichi Saito, Toshinori Hayashi
Life of this patent
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Abstract
The present invention provides an RNA amplification process, comprising steps in which double-stranded DNA containing a promoter sequence is produced using a first primer and second primer, at least one of which has the promoter sequence at the 5′ end, and reverse transcriptase, the double-stranded DNA is used as template for production of RNA transcript using RNA polymerase and the RNA transcript is used as template for subsequent DNA synthesis with the reverse transcriptase to produce the double-stranded DNA. The process yields an amplified RNA product which is assayed with a fluorescent intercalator dye-labeled nucleic acid probe.
Claims7
A method for assaying stanniocalcin-1 mRNA present in a sample, which method comprises
- (1) a step of producing double-stranded DNA which contains a promoter sequence and a specific base sequence within said mRNA downstream from said promoter sequence, using a first primer which is homologous to at least one section downstream from the 5′ end of said specific base sequence and a second primer which is complementary to at least one section upstream from the 3′ end of said specific base sequence, wherein said promoter sequence is at the 5′ end of either or both said first and second primers,
- (2) a step of producing RNA transcript using said double-stranded DNA as template,
- (3) a step of amplifying said RNA transcript in a chain-reaction manner using said RNA transcript as template for subsequent DNA synthesis, and
- (4) a step of measuring the amount of said RNA transcript.
An assay reagent for stanniocalcin-1 mRNA, characterized by containing as constituent elements oligonucleotides which have a combination of sequences consisting of a first primer comprising at least 15 contiguous bases of the sequence listed as SEQ ID NO:
- 1, and a second primer comprising at least 15 contiguous bases of the sequence listed as SEQ ID NO:
- 2, wherein a promoter sequence is at the 5′ end of either or both of said first and second primers.
An assay reagent for stanniocalcin-1 mRNA, characterized by containing as constituent elements oligonucleotides which are a combination of sequences consisting of a first primer comprising at least 15 contiguous bases of the sequence listed as SEQ ID NO:
- 1, a second primer comprising at least 15 contiguous bases of the sequence listed as SEQ ID NO:
- 2, wherein a promoter sequence is at the 5′ end of either or both of said first and second primers, and a fluorescent intercalator dye-labeled nucleic acid probe comprising at least 15 contiguous bases of the sequence listed as SEQ ID NO:
- 3 or the sequence complementary to said sequence.
Stanniocalcin-1 mRNA or a specific oligonucleotide which is complementary thereto, characterized by comprising at least 15 contiguous bases of the sequence listed as any one of SEQ ID NO:
- 1 to 3, or the sequences complementary to said sequences.
Dependencies are inferred from the claim text.