Gene probe for alveolar soft part sarcoma and application of gene probe kit

A soft tissue sarcoma and gene probe technology, applied in the application field of fluorescence in situ hybridization probes, can solve problems such as time-consuming inappropriate, complicated technology, difficult diagnosis, etc., and achieve accurate, reliable and simple diagnosis, strong fluorescent signal, and high accuracy high effect

Inactive Publication Date: 2015-01-28
THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

Alveolar soft tissue sarcoma with typical histological morphology is easier to diagnose, and some tumor tissues lack typical morphological manifestations, which need to be associated with paraganglioma, granular cell tumor, XP11.2 translocation/TFE3 gene fusion-related renal cancer and melanoma Differentiate different phases of tumors, especially in cases of rare disease sites, metastatic lesions, and few fine-needle aspiration biopsy tissues
Acinar soft tissue sarcoma forms an ASPL-TFE3 fusion gene, resulting in the overexpression of TFE3 protein. Therefore, TFE3 immunohistochemical detection has certain specificity for acinar soft tissue sarcoma, but in other tumors such as XP11.2 translocation/TFE3 gene Positive results of TFE3 immunohistochemistry can also be found in tissues such as fusion-related renal carcinoma, granulosa cell tumor, adrenocortical carcinoma, perivascular epithelioid cell tumor, and high-grade myxofibrosarcoma, so TFE3 immunohistochemistry can only be used as a diagnostic marker for glandular tumors. Auxiliary basis for vesicular soft tissue sarcoma
[0004] For alveolar soft tissue sarcoma, i

Method used

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  • Gene probe for alveolar soft part sarcoma and application of gene probe kit
  • Gene probe for alveolar soft part sarcoma and application of gene probe kit
  • Gene probe for alveolar soft part sarcoma and application of gene probe kit

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Step 1: Preparation of Gene Probes:

[0040] 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 fragments on the same side have a certain sequence overlap with each other. 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-2522M13 (chrX: 48420425-48602847, The fragment length is about 182Kb), RP11-416B14 (chrX: 485...

Embodiment 2

[0052] Example 2: Diagnostic kit for alveolar soft tissue sarcoma

[0053] Gene probe kit for alveolar soft tissue sarcoma, the kit is composed of probe hybridization solution and 4', 6-diamidino-2-phenylindole counterstaining agent, characterized in that:

[0054] (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-2312C1 and CTD-2248C21, labeled as green fluorescent signal;

[0055] (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 at -20°C in the dark;

[0056] (3) 4′,6-diamidino-2-phenylindole counterstain is mainly used for nuclear staining.

[0057] (4) The combination of RP11-634L10, RP11-51H16 and RP11-475F12 located on the ASPL gene of chromosome 17 described in the Gene Probe Kit for Acinar Soft Tissue Sarcoma, the three clone fragmen...

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Abstract

The invention relates to a gene probe for alveolar soft part sarcoma and application of a gene probe kit. Clonal fragments selected and used by the gene probe are RP11-634L10, RP11-51H16 and RP11-475F12 combination as well as CTD-2311N12, RP11-416B14, CTD-2522M13, CTD-2312C1 and CTD-2248C21 combination respectively. According to the gene probe, the defects of an influence on prognostic and postoperative treatment and the like due to a tedious and time-consuming RT-PCR (reverse transcription-polymerase chain reaction) and cell karyotype analysis method, limitation of RT-PCR application by RNA degradation and numerous other factors, and inaccurate diagnosis are overcome; the gene probe is high in accuracy, specificity and success rate, strong in fluorescent signal and easy and convenient to operate, and can be applied to paraffin sections; the specimen detection range is extended, a novel method for accurately, reliably, simply and conveniently diagnosing the alveolar soft part sarcoma is built, and a precedent for detecting the alveolar soft part sarcoma by FISH (fluorescence in situ hybridization) is created.

Description

technical field [0001] The invention belongs to the application field of fluorescent in situ hybridization probes, in particular to gene probes for alveolar soft tissue sarcoma and the application of kits thereof. Background technique [0002] Alveolar soft tissue sarcoma is a rare malignant soft tissue sarcoma of unknown origin that occurs in children and young adults. The main symptom is a soft tissue mass that grows slowly. Balanced translocation, namely: der(17)t(X;17)(p11.2;q25), resulting in translocation of the TFE3 telomeric side gene at Xp11.2 to 17q25 after duplication, and with the centromere of the 17q25ASPL gene Lateral fusions form the ASPL-TFE3 fusion gene. Most patients with alveolar soft tissue sarcoma are prone to distant metastasis after discovery and treatment, and are not sensitive to radiotherapy and chemotherapy. The overall prognosis is poor. The 5-year, 10-year, and 20-year survival rates are 60%, 38%, and 15%, respectively Therefore, accurate diag...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 甘卫东陈显成樊祥山屈峰曾浩郭宏骞
Owner THE AFFILIATED DRUM TOWER HOSPITAL MEDICAL SCHOOL OF NANJING UNIV
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