US 11,655,505 B2
Short-chain nucleic acid elongation primer set, assay kit, and short-chain nucleic acid elongation, amplification and detection methods
Keiko Ito, Kawasaki (JP); Mika Inada, Tokyo (JP); and Koji Hashimoto, Atsugi (JP)
Assigned to KABUSHIKI KAISHA TOSHIBA, Tokyo (JP)
Filed by KABUSHIKI KAISHA TOSHIBA, Tokyo (JP)
Filed on Feb. 10, 2021, as Appl. No. 17/172,378.
Application 17/172,378 is a continuation of application No. 16/017,367, filed on Jun. 25, 2018, granted, now 10,975,433.
Claims priority of application No. JP2017-135726 (JP), filed on Jul. 11, 2017; and application No. JP2018-083156 (JP), filed on Apr. 24, 2018.
Prior Publication US 2021/0207215 A1, Jul. 8, 2021
Int. Cl. C12Q 1/6876 (2018.01); C12P 19/34 (2006.01); C12N 15/10 (2006.01); C12Q 1/6848 (2018.01)
CPC C12Q 1/6876 (2013.01) [C12N 15/10 (2013.01); C12P 19/34 (2013.01); C12Q 1/6848 (2013.01); C12Q 2600/178 (2013.01)] 8 Claims
OG exemplary drawing
 
1. A primer set comprising:
a short-chain nucleic acid elongation primer set and
an amplification primer set,
wherein
the short-chain nucleic acid elongation primer set is configured to yield an elongated product from a target short-chain RNA containing a first sequence,
the elongated product is a mutually complementary double-stranded nucleic acid, one chain contains a second sequence, a third sequence, a fourth sequence, a complementary sequence of cDNA of the first sequence (hereinafter, “cDNA of the first sequence” referred to as “1′-th sequence”), fifth sequence and a sixth sequence in this order in a 3′ to 5′ direction,
the short-chain nucleic acid elongation primer set comprises:
a first elongation primer comprising a first elongation primer sequence which hybridizes with the first sequence, a complementary sequence of the fifth sequence and a complementary sequence of the sixth sequence in this order in the 3′ to 5′ direction; and
a second elongation primer comprising a second elongation primer sequence which hybridizes with the 1′-th sequence, the fourth sequence, the third sequence, and the second sequence in this order in the 3′ to 5′ direction, and
the amplification primer set is configured to amplify the elongated product by LAMP method which comprises;
the FIP primer comprising the second sequence and a complementary sequence of the third sequence in this order in the 3′ to 5′ direction,
the BIP primer comprising the complementary sequence of the sixth sequence and the fourth sequence in this order in the 3′ to 5′ direction, and
the loop primer comprising the complementary sequence of the 1′-th sequence.