Application of a multi-target gene parallel detection combination probe and its kit

Abstract

The invention discloses a combination probe for multi-target gene parallel detection and the application of the kit thereof, comprising a pair of stem-loop structure probes and a pair of double-strand hybridization structure probes; the stem-loop structure probes are 5 The complementary sequence at the 5' end of the 'oligonucleotide chain is connected to it by a C18 spacer to form a stem-loop structure probe with C18 as the ring and a single-stranded protruding 3' oligonucleotide; the double-stranded hybridization structure probe , including a long chain labeled with a fluorescent group at the 5' end and a short chain labeled with a quencher group at the 3' end. The full length of the short chain is partially complementary to the long chain to form a double-stranded complex. The multicolor fluorescence-labeled structural nucleic acid probe combination designed by the present invention can be used for the simultaneous detection of multiple target genes, with a sensitivity as high as 1 copy / microliter; combined with dual regulation of strand displacement thermodynamics and polymerase kinetics, the recognition resolution can reach At the single nucleotide level, the accurate identification of single nucleotide variations in multigene parallel detection is realized.

Description

Technical field [0001] The present invention belongs to the field of gene detection, and it is related to a multimodal gene parallel detection combination probe and a kit thereof. Background technique [0002] Multi-gene combination detection can acquire multiple gene sequences, sites, and abundance information at the same time, comprehensive acquisition characteristics, effectively improve the reliability and accuracy of analytical detection, and is of great significance in the fields of biological research and disease diagnosis. Multiple PCR amplification techniques can be widely used in the same reaction system to amplify multi-target sequences, which has been widely used in polynomic joint detection. However, multiple PCR technologies need to rely on accurate temperature-controlled thermal cycling devices, which is difficult to meet the analysis needs of simple, convenient molecular diagnosis technology. [0003] Nucleic acid isothermal amplification techniques can quickly ac...

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Problems solved by technology

[0004] However, in the existing detection methods, due to the many types and high concentrations of primers involved in the multiplex amplification system, the parallel detection of multiple targets has serious cross-interference, and the resolution of target sequence recognition is insufficient.

Therefore, it remains a challenge to develop methods that can accurately identify single-nucleotide variants in multigene parallel testing

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