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However, due to the intricate matrix composition of biological samples, there remains a need for further enhancement of sensitivity and specificity in current liquid biopsy methods to adequately meet clinical requirements.\u00a0Rolling circle amplification (RCA) is an efficient isothermal enzymatic reaction capable of rapidly producing numerous ultra\u2010long single\u2010stranded DNA molecules with repetitive sequences. By custom\u2010designing the DNA template sequence, RCA can generate functional products that integrate with various functional groups, achieving signal amplification and output with exceptional specificity and sensitivity in liquid biopsy. In this review, the fundamental principles and classifications of RCA are first introduced. 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