Improved fluorescence in-situ hybridization method and application thereof

Abstract

The invention provides an improved fluorescence in-situ hybridization method and an application thereof. The improved fluorescence in-situ hybridization method comprises the following steps: processing a specimen, conducting denaturation, processing a probe, conducting hybridization and conducting counter-staining. According to the improved fluorescence in-situ hybridization method provided by the invention, by conducting observation by virtue of a fluorescence microscope and calculating fluorescence hybridization signals of cells, conducting precise chromosome analysis on 81 patients with lymphoproliferative diseases and judging gene deletions and molecular genetic abnormalities of the patients, the following conclusions are obtained: molecular genetic analysis is conducive to diagnosis and differential diagnosis of lymphoid leukemia, and moreover, the molecular genetic analysis also serves as an important index for monitoring disease remission and recurrence and for judging a curative effect in the clinical field. On the basis of chromosomal karyotype analysis, the proper fluorescence in-situ hybridization probe and method are selected to conduct precise chromosome analysis on most lymphoid leukemia patients, and the various molecular genetic abnormalities are related to prognosis of the patients.

Description

technical field [0001] The invention relates to the technical fields of cytogenetics and molecular biology, in particular to an improved fluorescence in situ hybridization method and its application. Background technique [0002] In 2001, WHO carried out a new classification of lymphoid hematopoietic tissue tumors. Currently, clinically, lymphoproliferative diseases mainly refer to lymphocytic leukemia and some malignant lymphomas. Lymphocytic leukemia is a clonal malignant proliferation disease originating from B or T lineage lymphocytes, mainly including acute lymphocytic leukemia (ALL) and chronic lymphocytic leukemia (CLL). The former is characterized by abnormal proliferation and accumulation of immature prolymphocytes in blood, bone marrow, and lymphoid tissues. The latter is characterized by a chronic malignant proliferation of immunocompromised mature small lymphocytes in the blood, bone marrow, and lymphoid tissues, eventually leading to failure of normal hematopoi...

AI Technical Summary

Problems solved by technology

[0003] In the past, the evaluation indicators of diagnosis, treatment and prognosis mainly relied on the patient's clinical manifestations, peripheral blood, bone marrow cytomorphology, bone marrow biopsy, conventional cytogenetic (CC) studies, immune typing, etc. There are the following problems: First, although The quality and quantity of cell changes are an important basis for morphological diagnosis, but it is difficult to clearly distinguish between atypical prolymphocytes, prolymphocytes, myeloblasts and cells expressing myeloid and lymphoid markers at the same time; second, CC technology can only analyze metaphase cells, which are affected by the cell cycle and the number and quality of chromosomal division, which may lead to failure of karyotype analysis or missed detection of microclones with chromosomal aberrations, and the operation is time-consuming and laborious, and subjective and human factors have too much influence

Patients with lymphocytic disorders with p53 mutations have a poorer prognosis

FISH simultaneously detects P53, ATM deletion, RB1 deletion and +12 positive, indicating short overall survival and poor prognosis of CLL; positive GLP D13S25, indicating good prognosis of CLL patients; positive BCR / ABL1 fusion gene in ALL, indicating poor prognosis, but Can be treated with the targeted drug Gleevec; E2A-PBX1 positive is mainly seen in pre-B cell ALL, indicating a poor prognosis; TCR rearrangement is seen in T-ALL, and the positive prognosis is poor