Unicell detection method for tumor sample by means of mass spectrum flow system

Abstract

The invention provides a unicell detection method for a tumor sample by means of a mass spectrum flow system. The method comprises the following steps that (1) a cancer tissue biopsy sample or blood sample of a tumor patient is collected and processed to prepare a unicell suspension; (2) an antibody labeled with metal elements is used to mark the suspended unicell suspension with multiple targetsat the same time; (3) the marked unicell sample is detected by the mass spectrum flow system; and (4) on the basis of obtained mass spectrum flow type detection data, different immunocyte subgroups are clustered and analyzed, and expression levels of protein in tumor immunization detection points in different subgroup cells are quantitatively analyzed. Thus, the immucyte subgroups that can reflectthe clinical response degree of tumor immunotherapy can be accurately found in the immunocyte cells related to tumor of high heteroplasmy, the protein expression level threshold of the tumour immunization detection pointd can be determined appropriately, and a detection result is high in clinical correlation. Thus, the method has good clinical application prospects and higher social benefit.

Description

technical field [0001] The invention relates to a medical device, in particular to a single cell detection method, in particular to a single cell detection method for a tumor sample by using a mass spectrometer flow system. Background technique [0002] There is a complex tumor immune response in the tumor microenvironment. The body's immune system can recognize and eliminate existing tumor cells. The tumor immune response is mainly divided into seven steps: 1. Release tumor antigens; 2. Present tumor antigens; 3. Activate Effector T cells; 4. T cells transfer to tumor tissue; 5. T cells infiltrate tumor tissue; 6. T cells recognize tumor cells; 7. T cells clear tumor cells. However, different tumor cells can inhibit the immune response of the human body through different methods, thereby avoiding the killing of immune cells, resulting in immunosuppression, immune tolerance, and promoting the occurrence and progression of tumors. This characteristic of tumor cells is called ...

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Problems solved by technology

However, due to the high degree of heterogeneity of cells in cancer tissue, IHC cannot detect at the single-cell level. Uniformity, the defect that the positive threshold is difficult to determine, and the correlation of the prediction results is not high; and it is impossible to detect multiple targets at the same time, and it is impossible to simultaneously detect multiple tumor immune targets in precious tumor biopsy samples

[0009] In addition, although the gene detection method based on high-throughput sequencing can achieve the detection of multiple target genes at the single-cell level and overcome the heterogeneity of tumor cells, the detection at the gene level is an indirect method, which is based on the degree of gene variation. Probabilistic prediction of clinical response, and the difference at the gene level is not completely correlated with the difference at the protein level or cell phenotype level, so it has a high false positive rate and false negative rate; and there are a large number of detection results For mutations with unknown functions, massive information has very high requirements for data processing; in addition, the cost is high, and it is inconvenient to perform multiple dynamic monitoring to evaluate the prognosis of tumors

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