Digital PCR-based vector biological nucleic acid quantitative detection method

Abstract

The invention discloses a vector biological nucleic acid quantitative detection method based on digital PCR, belongs to the technical field of vector biological detection, and in the method, the comprehensive performance of vector biological nucleic acid detection is remarkably improved through multi-dimensional optimization. In a primer concentration adjusting link, a fluorescence signal feedback mechanism is adopted to realize dynamic balance, the amplification difference of each target is accurately captured through real-time signal monitoring in a pre-amplification stage, and the primer concentration is adjusted in a stepped manner according to the signal deviation degree, so that the concentration of the high-abundance target primer is reduced to avoid resource preemption; and increasing the concentration of the low-abundance target primer to enhance amplification power. Meanwhile, sequence homology analysis is introduced, and a primer combination with relatively high homology is subjected to coordinated regulation, so that non-specific amplification caused by pure increase of concentration is avoided, and competitive inhibition among primers is reduced through cross regulation, so that multiple targets reach dynamic balance of amplification efficiency in the same reaction system; and the common amplification bias problem in the traditional multiplex PCR is effectively solved.

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