Method for discriminating non-blood-borne nucleated cells enriched from human or animal biological fluid

Abstract

The invention provides a method for carrying out fast effective separation of red blood cells and non-blood-borne nucleated cells from human biological fluid. According to the method, red blood cell cracking and melting are not needed, and no influence is caused upon target cells (such as tumor cells in the body fluid) in human body fluid, such that original cell morphology is well maintained, and subsequent cell identification and determination are facilitated. The invention also relates to a method for discriminating non-blood-borne nucleated cells (such as tumor cells, vascular endothelial cells, and the like) enriched from human or animal biological fluid. According to the invention, anti-human leukocyte antibody immune-fluorescence staining and chromosome fluorescence in-situ hybridization are carried out concurrently, such that blood-borne nucleated cells are discriminated, and blood-borne nucleated cell interference during a tumor cell determination process is effectively eliminated. Therefore, non-blood-borne epithelial-source (solid tumor) or non-epithelial-source (melanoma) rare tumor cells can be enriched, extracted, and effectively discriminated from human body fluid rapidly with high efficiency.

Description

technical field [0001] The present invention mainly relates to the identification of tumor cells of malignant epithelial origin (solid tumors) or non-epithelial origin (melanoma) enriched from human or animal biological fluids using simultaneous immunofluorescence in situ hybridization. Background technique [0002] The huge reality and potential clinical significance of extracting and detecting malignant epithelial cells (tumor cells) in human body fluids (pleural effusion or pleural effusion, ascites or peritoneal effusion, cerebrospinal fluid, lymph fluid and / or blood, etc.) confirmed. In many collected human biological fluid samples, a large number of red blood cells and white blood cells are often mixed, which greatly interferes with the experimental operation and the judgment of the results. At present, the common method of removing red blood cells in clinical or medical experiments is to use hypotonic lysis However, in the process of hypotonic lysis, along with the c...

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Problems solved by technology

[0004] In addition, although some articles reported that the asynchronous FISH method based on tumor epithelial cell staining was used to detect circulating tumor cells in blood, that is, FISH and anti-tumor cell marker antibody staining were performed asynchronously and alternately, but considering A large number of shed solid tumor tumor cells lost the characteristics of tumor epithelial cells due to the complete change of the growth environment in the liquid environment, which directly affected the antibody immunostaining of tumor cells, resulting in a large number of false negatives, so there is no clinical significance. use value