DNA probe library for hybridization with MSI (micro-satellite instability) related micro-satellite sites, detection method and kit

Abstract

The invention provides a DNA probe library for hybridization with MSI (micro-satellite instability) detection related micro-satellite sites. The DNA probe library comprises one or more DNA probes capable of hybridizing with the MSI state related microsatellite sites. In the DNA probes, the MSI related micro-satellite sites are shown in the following sequences: SEQ ID NO. 1-66. Besides, the invention provides a method for enriching and detecting the MSI related micro-satellite sites by the probe library. With combination of the method with the next generation sequencing technology (NGS), sensitivity, accuracy and comprehensiveness of MSI detection can be greatly improved.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a method for enrichment and detection of microsatellite sites related to microsatellite instability (MSI). This method can accurately enrich the specific fragments of MSI-related microsatellite loci, and the obtained DNA sample library can be further combined with next-generation sequencing technology (NGS), and quantitatively evaluate the patient's MSI status through bioinformatics analysis, providing a basis for the diagnosis of tumors. Prognosis, and the design of clinical treatment plan to provide guidance and theoretical basis. Background technique [0002] Microsatellite instability (Micro-Satellite Instability, MSI) refers to the change in the length of microsatellite alleles due to abnormal insertion or removal of repetitive sequences during DNA replication, and the change is due to various reasons (such as DNA mismatch repair The mechanism of gene promoter methylation in...

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Problems solved by technology

However, the common PCR method has problems such as cumbersome operation procedures, long time-consuming, low sensitivity, and high uncertainty of detection results. In multiplex PCR detection, the interference relationship between different primers is very complicated, and the amplified product is highly mixed. There are high requirements on the selection and concentration of primers, these factors will undoubtedly greatly increase the detection cost

For the detection of DNA mismatch repair gene defects, traditional gene sequencing methods, such as Sanger sequencing, also have problems such as high cost, low throughput, and low precision

However, the method of immunohistochemistry has low specificity and reproducibility, requires high sample quality, and the operation is relatively complicated.

Finally, due to the limited number of microsatellite loci that can be detected by existing conventional MSI detection methods, it is difficult to distinguish high-frequency MSI-H from low-frequency MSI-L

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