Method for negatively removing hematuria leukocyte-enriched epithelial cells based on magnetic beads method

Abstract

The invention belongs to the field of a method for sampling, inoculating or diverging samples, a method for physically separating intact cells, and discloses a method for negatively removing hematurialeukocyte-enriched epithelial cells based on a magnetic beads method. The method for negatively removing hematuria leukocyte-enriched epithelial cells based on a magnetic beads method comprises the following steps: adding a red blood cell lysate for 10 min; centrifuging 1500 g for 10 min, and removing a supernatant; adding 20 microliters of CD45 magnetic beads to the cell pellets, uniformly mixing, and incubating at room temperature for 1 hour; treating in the magnetic field for 5 min, and aspirating the liquid into an EP tube. Through the technology of introducing the magnetic beads to remove leukocyte, the sensitivity of FISH technology for detecting urine exfoliated cells for diagnosis of bladder cancer can be improved from 80% to 90%, which has important clinical significance.

Description

technical field [0001] The invention belongs to the field of methods for sampling, inoculating or diffusing samples; methods for physically separating intact cells, and in particular to a method for negatively removing hematuria leukocyte-enriched epithelial cells based on a magnetic bead method. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: The diagnosis of bladder cancer and ureteral cancer often relies on cystoscopy or ureteroscopy and pathological or histological examination, and the patient's compliance is not high. Traditional imaging (ultrasound, CT, etc.) often finds tumors in the middle and late stages, and the treatment effect is not good , the prognosis is poor. How to make early diagnosis and recurrence monitoring of urinary system transitional cell carcinoma has become an urgent problem to be solved. Bladder cancer has certain chromosomal aberrations during its occurrence and development. Dome...

AI Technical Summary

Problems solved by technology

FISH exfoliated cell chromosome examination has low sensitivity and high specificity. Non-invasive urine FISH examination for the early diagnosis of bladder cancer and ureteral cancer still has the following problems: urine is often interfered by blood cells, which limits the sensitivity and specificity of the detection method. It is necessary to establish a stable new technology for removing urine blood cells and enriching epithelial cells

[0003] In summary, the problems in the prior art are: Urine is often disturbed by blood cells, and the increase of blood cells reduces the proportion of epithelial cells in all cells. If epithelial cells are not enriched, after pretreatment and fluorescence in situ hybridization, when interpreting under a fluorescent microscope , most of the cells observed in each field of view are white blood cells, it is difficult to see epithelial cells, and the insufficient number of epithelial cells makes it difficult to observe positive cells in epithelial cells, that is, false negatives may occur, which reduces the detection of FISH methods Susceptibility to chromosomal abnormalities in exfoliated cells

[0004] The difficulty and significance of solving the above technical problems: In order to enrich more epithelial cells for subsequent FISH detection, it is difficult to use the positive enrichment method, because there are few specific markers for urinary system epithelial cells, and positive markers cannot be used for enrichment; Positive cells can be easily removed at the same time by flow cytometry etc.

Images