发明名称 |
MODIFIED STEM-LOOP OLIGONUCLEOTIDE MEDIATED REVERSE TRANSCRIPTION AND BASE- SPACING CONSTRAINED QUANTITATIVE PCR - PCR |
摘要 |
There is provided a method for detecting a target RNA molecule in a sample. The method comprises reverse transcribing the target RNA contained in the sample using an RT oligonucleotide, the RT oligonucleotide comprising a stem-loop portion containing one or more nucleotides modified or modifiable to block DNA polymerase extension and a target annealing portion that is complementary to a downstream portion of the target RNA, the target annealing portion located 3' to the stem-loop portion, to produce a reverse transcription product that comprises the RT oligonucleotide and a 3' extended region; amplifying the reverse transcription product using (i) a first amplification primer that anneals to a downstream portion of the 3' extended region of the reverse transcription product and (ii) a second amplification primer that anneals to an interface portion of a DNA strand complementary to the reverse transcription product, the interface portion comprising a region that is complementary to a 3' portion of the RT oligonucleotide and a 5' portion of the 3' extended region in the reverse transcription product, to produce an amplification product; and detecting the amplification product; wherein the stem-loop portion adopts a stem-loop structure under conditions used for said reverse transcribing but does not adopt the stem-loop structure under conditions used for said amplifying and wherein when the stem-loop portion contains one or more nucleotides that are modifiable to block DNA polymerase extension, the method further comprises modifying the modifiable nucleotide prior to said amplifying. |
申请公布号 |
HK1187378(A1) |
申请公布日期 |
2016.08.26 |
申请号 |
HK20140100278 |
申请日期 |
2014.01.10 |
申请人 |
National University of Singapore |
发明人 |
TOO, Heng, Phon;ANWAR, Azlinda, B. B |
分类号 |
C12Q |
主分类号 |
C12Q |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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