Aspl‑tfe3 fusion renal cancer gene probe and its kit application

A gene probe, kidney cancer technology, applied in the application field of fluorescence in situ hybridization probes, can solve the problems of inability to accurately diagnose kidney cancer, restrict RT-PCR, and be unsuitable for wide application, and achieve high success rate and rapid diagnosis. , the effect of simple diagnosis

Inactive Publication Date: 2018-01-05
THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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AI Technical Summary

Problems solved by technology

For karyotype analysis, tumor cells must be cultured to the mitotic stage, and then G-banding and other techniques are used to analyze the karyotype of tumor cells. This method is cumbersome and time-consuming and is not suitable for clinical application.
Similarly, considering the rarity of ASPL-TFE3 fusion RCC and the complexity of the PCR test process, it is impossible to routinely use RT-PCR to detect ASPL-TFE3 fusion genes in all RCC tissues, and when ASPL-TFE3 fusion is suspected In advanced renal cancer, RNA can only be extracted from paraffin sections. The rapid degradation of RNA and many other factors limit the application of RT-PCR, and the technology is complicated and not suitable for wide application.
These limitations lead to the inaccurate and timely diagnosis of ASPL-TFE3 fusion RCC, which affects the judgment of clinicians on the prognosis and postoperative management of patients with ASPL-TFE3 fusion RCC

Method used

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  • Aspl‑tfe3 fusion renal cancer gene probe and its kit application
  • Aspl‑tfe3 fusion renal cancer gene probe and its kit application
  • Aspl‑tfe3 fusion renal cancer gene probe and its kit application

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Experimental program
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Embodiment 1

[0041] Step 1: Preparation of Gene Probes:

[0042] Find the bacterial artificial chromosome (BAC clone) corresponding to the X chromosome TFE3 gene and ASPL gene through http: / / genome.ucsc.edu / , select the corresponding BAC clone fragments respectively, and control the complete coverage of the BAC clone fragments selected on the same side relative to the gene , and the ipsilateral fragments have a certain sequence overlap with each other or the distance is less than 10Kb. According to the above requirements, the BAC clone fragments on the ASPL gene were selected as RP11-634L10 (chr17: 79796813-79969288, the fragment length is about 172Kb), RP11-51H16 (chr17: 79931578-80097452, the fragment length is about 166Kb) and RP11-475F12 ( chr17: 80032268-80216871, the fragment length is about 185Kb); the BAC clone fragment on the TFE3 gene is CTD-2311N12 (chrX: 48713289-48923401, the fragment length is about 210Kb), CTD-2516D6 (chrX: 48265836-48484403, The fragment length is about 21...

Embodiment 2

[0054] Example 2: ASPL-TFE3 Fusion Kidney Cancer Diagnostic Kit

[0055] Gene probe kit for ASPL-TFE3 fusion kidney cancer, the kit is composed of probe hybridization solution and 4', 6-diamidino-2-phenylindole counterstaining agent, characterized in that:

[0056] (1) The combination of RP11-634L10, RP11-51H16, and RP11-475F12 located on the ASPL gene on chromosome 17, marked with a red fluorescent signal; Combination of CTD-2516D6, CTD-2312C1, CTD-2248C21 and RP11-959H17, labeled with green fluorescent signal;

[0057] (2) The probe hybridization solution is prepared by mixing the probe with Human Cot-1 DNA, hybridization buffer, and purified water in proportion, and it needs to be stored in a dark place at -20°C;

[0058] (3) 4′,6-diamidino-2-phenylindole counterstain is mainly used for nuclear staining.

[0059] The combination of RP11-634L10, RP11-51H16 and RP11-475F12 located on the ASPL gene of chromosome 17 is marked with a red fluorescent signal; located on the TFE3...

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Abstract

The invention relates to an ASPL-TFE3 fusion kidney cancer gene probe and the application of the kit. The clone fragments selected for gene probes in the present invention are respectively RP11‑634L10, RP11‑51H16 and RP11‑475F12 combinations, and CTD‑2311N12, RP11‑416B14, CTD‑2522M13, CTD‑2516D6, CTD‑2312C1, CTD‑2248C21 and RP11‑959H17 combination. The invention overcomes the defects that the RT-PCR and cell karyotype analysis methods used in the past are cumbersome and time-consuming and are not suitable for clinical application. The present invention uses FISH technology to detect the unique ASPL-TFE3 fusion gene in ASPL-TFE3 fusion kidney cancer to diagnose the tumor. The application of the gene probe has high accuracy, high specificity, high success rate, strong fluorescent signal, and simple operation. , rapid diagnosis, can be applied to paraffin sections, expanded the scope of detection specimens, established a new method for the accurate, reliable and simple diagnosis of ASPL-TFE3 fusion renal carcinoma, and pioneered the detection of ASPL-TFE3 fusion renal carcinoma by FISH.

Description

technical field [0001] The invention belongs to the application field of fluorescent in situ hybridization probes, in particular to the application of ASPL-TFE3 fusion kidney cancer gene probes and kits thereof. Background technique [0002] Xp11.2 translocation / TFE3 gene fusion-related renal cell carcinoma is more common in adolescents, accounting for about 1 / 3 of renal cancer in children, and 15% of renal cancer under the age of 45. The prognosis is poor, and the prognosis of adult patients is worse than that of children. It is mainly related to pathological type, tumor grade and stage, whether there is distant metastasis, etc. Xp11.2 translocation / TFE3 gene fusion correlation is not effective for conventional immunotherapy, and it is not sensitive to radiotherapy and chemotherapy. Surgery is the main treatment method, and patients are prone to lymph node and distant metastasis. Only targeted therapy has been proved to have a certain effect. effect. [0003] ASPL-TFE3 fu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 甘卫东陈显成何健杨军郭宏骞贾瑞鹏
Owner THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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