METHOD FOR ENRICHING AND DETECTING LOW-ABUNDANCE MUTANT DNA
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United States of America
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Apr 3, 2025
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Sep 14, 2024
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Sep 14, 2024
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Abstract
The invention relates to a method for enriching and detecting low-abundance mutant DNA. The invention provides a wild-type amplification blocker, wherein the Tm value (Tm1) of the wild-type amplification blocker specifically binding to a wild-type template is greater than the Tm value (Tm2) of the wild-type amplification blocker specifically binding to a mutant template, and the 3′ terminus of the wild-type amplification blocker has a blocking group. The invention further provides a corresponding amplification refractory mutation system and a corresponding nucleic acid detection system, wherein the amplification refractory mutation system comprises a probe (the Tm value of the probe binding to the wild-type template is Tm3, and Tm1>Tm3>Tm2) of a template competitively binding to the wild-type amplification blocker, and forward and reverse primers. According to the technical solution of the invention, the system can enrich low-abundance DNA mutations and detect multiple types of base mutations.
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