A DNA probe library, detection method and kit for hybridizing microsatellite loci associated with microsatellite instability

Abstract

The present invention provides a library of DNA probes for hybridization to microsatellite loci associated with microsatellite instability (MSI) detection, said DNA probe library comprising one or more microsatellite loci capable of correlating with MSI status Spot the DNA probe for hybridization. Among the DNA probes, the MSI-related microsatellite site probes are shown in the following sequence: SEQ ID NO.1-66. In addition, the present invention provides a method for enriching and detecting MSI-related microsatellite loci using the probe library. Combining this method with next-generation sequencing technology (NGS) can greatly improve the sensitivity and accuracy of MSI detection and comprehensiveness.

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 inhi...

AI Technical Summary

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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