摘要:
Disclosed is a method of amplifying and detecting cytokeratin 19 mRNA in RNA amplification process, comprising: a step for forming a double-stranded DNA containing a promoter sequence with a reverse transcriptase by use of a combination of oligonucleotides consisting of a first primer having a sequence homologous to a portion of cytokeratin 19 mRNA and a second primer having a complementary sequence, wherein the promoter sequence is added to the 5'-end of either the first primer or the second primer, forming an RNA transcription product by use of an RNA polymerase with using the double-stranded DNA as template, and forming the double-stranded DNA by use of a reverse transcriptase by continuing to use the RNA transcription product as a template of DNA synthesis, by measuring an amount of amplified RNA produced over time with an oligonucleotide probe designed so that signal properties change with the formation of a complementary double strand with the amplified RNA.
摘要:
Disclosed herein is a method for highly sensitive determination of a target substance by means of an aptamer. The method includes causing an aptamer capable of specific binding with a target substance to bind competitively with said target substance and a nucleic acid strand having a base sequence complementary to at least a portion of said target substance, detecting at least either of the physical change and the chemical change that results from said aptamer binding with said nucleic acid strand, and determining said target substance based on the result of detection.
摘要:
PROBLEM TO BE SOLVED: To provide the development of reliable diagnostic tests to detect hepatitis A virus in viremic samples.SOLUTION: Hepatitis A virus-specific primers and probes derived from conserved regions of the hepatitis A virus genome are disclosed. Nucleic acid-based assays using the capture oligonucleotides, primers and probes are also disclosed. An isolated oligonucleotide not more than 60 nucleotides in length is disclosed and the isolated oligonucleotide comprises: (a) a nucleotide sequence of at least 10 continuous nucleotides from a specific sequence; (b) a nucleotide sequence having 90% sequence identity to a nucleotide sequence of (a); or (c) complements of (a) and (b).