METHOD FOR ANALYZING TEMPLATE NUCLEIC ACID

WIPO Patent Application WO/2016/140070

A1

Provided is an analysis method for analyzing a template nucleic acid at a high accuracy. The method for analyzing a template nucleic acid comprises: an amplification step for amplifying a target sequence in the template nucleic acid and a complementary sequence to the target sequence; and a detection step for detecting association of a probe with an amplification product obtained in the above amplification step or dissociation of the associated product accompanying temperature change. In the amplification step, a primer set, said primer set consisting of a first primer from the 3'-end of which a polymerase initiates priming to synthesize the target sequence and a second primer from which the polymerase synthesizes the complementary sequence, is used. Between the first and second primers, a primer initiating the synthesis of the sequence hybridizable with the probe is a turnback primer containing a primer region and a probe region, wherein the probe region is directly or indirectly bound to the 5'-end of the primer region, and the other primer is an arbitrary primer that is capable of enabling priming of the polymerase from the 3'-end thereof. In the amplification step, the turnback primer extends the target sequence or complementary sequence from the primer region. In the detection step, the turnback primer also serves as the probe. In the amplification product obtained in the previous step, the probe region of the turnback primer is folded back toward the extended target sequence or complementary sequence within the molecule thereof to thereby detect the association of the probe region with the target sequence or complementary sequence.

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TSUJIMARU KOICHIRO (JP)

PCT/JP2016/054718

September 09, 2016

February 18, 2016

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DNAFORM KK (JP)

C12N15/09; C12M1/00; C12Q1/68; G01N21/78

JP2014515268A	2014-06-30
JP2007135489A	2007-06-07

THELWELL N. ET AL.: "Mode of action and application of Scorpion primers to mutation detection.", NUCLEIC ACIDS RES., vol. 28, no. 19, 2000, pages 3752 - 3761, XP002247004, DOI: doi:10.1093/nar/28.19.3752

NAKAYAMA YUMI (JP)
Yumi Nakayama (JP)

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