Abstract: The present invention discloses a method for detecting the mutation and methylation of tumor-specific genes in ctDNA, and this method can simultaneously detect the mutation (including point mutation, insertion-deletion mutation, HBV integration and other mutation forms) and/or methylation of tumor-specific genes in ctDNA in one sample. Not only the sample size requirement is low, but the MC library prepared by this method can support 10-20 subsequent detections. The results of each test can represent the mutation status of all the original ctDNA specimens and the methylation modification status of the region covered by the restriction sites, without reducing the sensitivity and specificity. The present invention has important clinical significance for early tumor screening, disease tracking, efficacy evaluation, prognosis prediction and the like, and has great application value.

Abstract: A method for preparing an amplicon library for detecting the variation in a region to be tested of a target gene of a sample, including the following steps: 1) designing and synthesizing a forward outer primer F1, a forward inner primer F2, and a reverse primer R according to the target region; 2) carrying out a one-step PCR amplification on the sample to be tested using the forward outer primer F1, the forward inner primer F2, and the reverse inner primer R to obtain an amplified product, i.e., the amplicon library of the target region. This one-step library preparation technology can be applied to all second-generation platforms including IonTorrent, illumina and BGI/MGI platforms. Based on the library preparation method, the present invention has developed detection products for SNP, Ins/Del, CNV and methylation of DNA, as well as detection products for s gene fusion and expression of RNA samples.

Abstract: The present invention provides a kit for early screening of hepatocellular carcinoma, comprising a gene marker detection reagent and a protein marker detection reagent. The invention also provides a preparation method and application of the kit. The kit comprising specific gene markers and protein markers of the present invention has been demonstrated to be effective in achieving early screening of HCC in community populations, particularly in prospective studies.

Abstract: A method of ctDNA library construction and sequencing data analysis for simultaneously detecting multiple common mutations in liver cancer. The library construction method and sequencing data analysis process have the following advantages: 1. Simultaneous detection of multiple mutation forms in liver cancer without capturing; 2. Suitable for efficient capture of ultra-small target regions; 3. The library may support 10-20 tests; 4. Ligate the DNA barcode to the starting ctDNA molecule during the library construction process, and cooperate with the biological information analysis process to achieve high specific detection of low-frequency mutations in ctDNA; 5. The library is usable for PCR hot spots detection and sequencing by a capture method at the same time, the added DNA barcode may effectively filter out false positive mutations and achieve high-specificity sequencing based on duplex.

Abstract: The present invention discloses a method for rapidly constructing amplicon library including the following steps: 1. Synthesizing a primer combination for constructing an amplicon library of a DNA sample, the primer combination of the amplicon library used to construct the DNA sample includes: a forward fusion primer designed according to the target amplicon, a reverse fusion primer designed according to the target amplicon, a forward universal primer and a reverse universal primer; 2. Constructing a PCR reaction system for the DNA sample; 3. Performing PCR. The method according to the present invention can be used to construct an amplicon library in a simple and rapid manner, and since a barcode is introduced before the start of PCR, the possibility of cross-contamination between the sample and the library is greatly reduced.

Abstract: The present invention discloses a method for rapidly constructing amplicon library including the following steps: 1. Synthesizing a primer combination for constructing an amplicon library of a DNA sample, the primer combination of the amplicon library used to construct the DNA sample includes: an upstream fusion primer designed according to the target amplicon, a downstream fusion primer designed according to the target amplicon, an upstream universal primer and a downstream universal primer; 2. Constructing a PCR reaction system for the DNA sample; 3. Performing PCR. The method according to the present invention can be used to construct an amplicon library in a simple and rapid manner, and since a barcode is introduced before the start of PCR, the possibility of cross-contamination between the sample and the library is greatly reduced.