Positive quality control product for detecting HBB/GJB2/ATP7B/PAH genetic disease genes and preparation method of positive quality control product

Abstract

The invention relates to a positive quality control product for detecting HBB / GJB2 / ATP7B / PAH genetic disease genes and a preparation method of the positive quality control product. A DNA sequence, which includes four mutant gene segments, namely HBB_CD41-42, GJB2_235delC, ATP7B_2333G>T and PAH_728G>A, is synthesized, and the sequence is interpolated into a plasmid vector and is mixed with wild-type human genome DNA according to a copy number of 2 to 1, so that the positive quality control product is obtained. The positive quality control product prepared by the invention, as a positive internal standard quality control material of a kit for detecting HBB / GJB2 / ATP7B / PAH gene mutation related to genetic diseases, can be used for accurately monitoring an entire experimental process of detecting the related genetic diseases with the adoption of the detection kit; therefore, the positive quality control product can serve as a standard for judging whether an experiment is successful or not.

Description

[0001] Field [0002] The present invention relates to the field of molecular biology and medical detection, and relates to a common genetic disease based on a high-throughput sequencing platform—a positive quality control product for gene detection of thalassemia, deafness, phenylketonuria and hepatolenticular degeneration. its preparation method. Background technique [0003] Genetic diseases refer to the abnormal traits of the offspring caused by the abnormality of the genetic material (chromosome, DNA) in the fertilized egg or the abnormality of the genetic information carried by the germ cells. Genetic diseases are divided into single-gene genetic diseases and polygenic genetic diseases. There are many types of single-gene genetic diseases with high incidence, which not only cause serious harm to the health of patients, but also bring heavy mental and economic burdens to families and society. burden. Therefore, it is very necessary to pass the genetic testing of early n...

AI Technical Summary

Problems solved by technology

[0010] There are two methods for preparing positive quality control products in traditional genetic disease detection kits: 1. Directly dilute the recombinant plasmid with a single PCR product positive for mutations to 10 3 Copies / μL is used as a positive quality control product. The quality control product prepared by this method can only cover the insert fragment containing the specific mutation site, but other regions of genomic DNA cannot be covered; 2. Two or more mutation positive The recombinant plasmids of the PCR products were mixed in proportion and diluted to 10 3 Copies / μL is used as a positive quality control substance. Although this method can simulate the detection of two or more mutation sites, it cannot cover other regions of genomic DNA; neither of the above two quality control substances can truly simulate human genomic DNA.

For the modern high-throughput sequencing technology that can sequence hundreds of thousands to millions of DNA molecules at a time, the sequencing of quality control products prepared by mixing single or several PCR products is obviously not suitable.

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