Method for detecting mutation information in multiplex amplification sequencing product of genome

Abstract

The invention discloses a method for detecting mutation information in a multiplex amplification sequencing product of a genome. The method comprises steps as follows: sequencing data are subjected to quality assessment and preprocessing; a recognizable sequencing sequence is selected for sequence assembling; the recognizable sequencing sequence or a sequence obtained through assembling is compared with a reference gene sequence, and preliminary variation information is obtained; fine calibration of sequence variation is performed according to different types of conditions; a calibrated sequencing fragment is obtained; the homozygosis or heterozygosis state of a target fragment is obtained according to the type of the sequencing fragment with the highest abundance; finally, the mutation information in the multiplex amplification sequencing product of the genome is obtained. By means of the method, the amplification product can be rapidly, efficiently and accurately recognized, and the calculation resources are saved; the sequence assembling process is compatible, and the problem of reduction of the quality value of basic groups produced in the sequencing process can be effectively solved; the homozygosis / heterozygosis state of variation information can be more effectively and stably judged, and random errors introduced in the PCR (polymerase chain reaction) process and the sequencing process are eliminated.

Description

technical field [0001] The invention relates to the field of gene sequencing, in particular to a method for detecting mutation information in genome multiple amplification sequencing products. Background technique [0002] At present, there are many methods for detecting mutations in gene sequences. Among them, specific amplification of a specific target region through multiple amplification and high-throughput sequencing of the amplified products is a preferred method with high efficiency, economy and high accuracy. This type of method can expand the detection throughput of traditional single-point PCR. At the same time, high-throughput sequencing technology can obtain accurate digital signals of the target region, and can more accurately describe the sequence variation. [0003] However, such methods will generate a large amount of sequencing sequence information during the detection process, requiring professional data analysis methods to interpret the sequencing results...

AI Technical Summary

Problems solved by technology

However, due to the need to compare the sequencing sequence to the whole genome region, this type of method runs slowly and requires high resource consumption.

Moreover, in the process of mutation detection, the statistics of mutations are carried out at the level of a single site. Errors generated during PCR or sequencing can only be eliminated through probability model calculations, which are susceptible to interference

Furthermore, since the method is positioned as a whole-genome comparison, the same processing method is adopted for the comparison of each gene position, and the difference in expression of different genes is not considered, which leads to inability to match the biological significance in the later functional annotation. Direct association with potential impact on genetic counseling of tested samples

[0004] In summary, at present, there is no efficient, fast and accurate analysis method for this kind of data in clinical practice.

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