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Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof

An xp11.2, probe technology, applied in the application field of fluorescence in situ hybridization probes, can solve the problem of different sizes of cloned fragments, and achieve the effect of improving accuracy and reliable success rate

Inactive Publication Date: 2012-04-25
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] Evaluation: The size of the cloned fragments of this probe is different, and the difference between the largest and the smallest is more than 300 kb, which is easy to cause heterogeneity of in situ hybridization conditions

Method used

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  • Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof
  • Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof
  • Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1: the preparation of DNA probe combination:

[0045] Select two BAC clone fragments that can be connected at both ends of the X chromosome TFE3 gene, control the farthest distance between the probes at both ends within 1500kb, keep a certain distance between the BAC clone fragments without overlapping, and the fragment sizes are similar. The cloned fragments were obtained from the Human BAC Cloning Center of EmpireGenomics (http: / / www.empiregenomics.com / helixhq / clonecentral / search / human). The BAC fragments on the centromere side of TFE3 were RP11-58H17 (200kb) and RP11-352D11 (175kb), and the BAC fragments on the telomeric side of TFE3 were RP11-416B14 (182kb) and RP11-107C19 (160kb). The linking sequence of BAC cloned fragment and TFE3 gene is RP11-58H17, RP11-352D11, TFE3, RP11-416B14, RP11-107C19. The positioning structure of the probe assembly is as follows figure 1 shown. Use the gap translation method to label the two BAC clone fragments on the telo...

Embodiment 2

[0047] Embodiment 2: fluorescence in situ hybridization process:

[0048] 1. Tissue microarray construction of specimens:

[0049] A total of 51 cases of renal cell carcinoma diagnosed in Nanjing General Hospital of Nanjing Military Command were collected. Two experienced pathologists refer to WHO 2004 urinary and male reproductive system classification criteria, immunohistochemical TFE3 positive, and RT-PCR detection of fusion gene results ( figure 2 A, B, C, 3 of the 51 patients were diagnosed at the mRNA level, and the correspondence with the results of this experiment can better reflect the reliability of this experiment), re-diagnosed and evaluated, and the final diagnosis of Xp11.2 was easy 24 cases of dislocated renal cell carcinoma, 9 cases of clear cell carcinoma, 17 cases of papillary renal cell carcinoma (16 cases of sporadic papillary renal cell carcinoma, 1 case of papillary renal cell carcinoma associated with VHL disease), 1 case of unclassified renal cell car...

Embodiment 3

[0062] Example 3: Xp11.2 Translocation Kidney Cancer Diagnostic Kit

[0063] The kit contains the probe combination described in Example 1, and the characteristics of the probe combination are mainly:

[0064] (1) The BAC clone probes on the centromere side of TFE3 are RP11-58H17 (fragment length 200kb) and RP11-352D11 (fragment length 175kb), labeled with green fluorescence. These two probes show a green fluorescent signal under the fluorescence microscope, representing the centromere side of the TFE3 gene.

[0065] (2) The BAC clone probes on the telomeric side of TFE3 are RP11-416B14 (fragment length 182kb) and RP11-107C19 (fragment length 160kb), labeled with red fluorescence. These two probes show a red fluorescent signal under the fluorescent microscope, representing the telomeric side of the TFE3 gene.

[0066] (3) Under normal circumstances, the red and green signals are connected or overlapped to form a fusion signal. In Xp11.2 translocation RCC, the red and green s...

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Abstract

The invention discloses a probe combination for diagnosing Xp11.2 translocation renal cancer and an application thereof, which belong to the field of fluorescent in situ hybridization probe application. The probe combination is BAC cloning fragments RP11-58H17, RP11-352D11, RP11-416B14 and RP11-107C19. The specificity and the sensibility of the Xp11.2 translocation renal cancer diagnosis by the probe combination respectively reach 100 percent, convenience, high speed and reliability are realized, in addition, the success ratio is high, and the probe combination can be used for preparing Xp11.2 translocation renal cancer diagnosis reagents.

Description

technical field [0001] The invention belongs to the application field of fluorescent in situ hybridization probes, and relates to a probe combination for diagnosing Xp11.2 translocation renal cancer and its application in preparing diagnostic reagents for Xp11.2 translocation renal cancer. Background technique [0002] In 2004, WHO revised the histopathological classification of RCC in 1997, adding Xp11.2 translocation / TFE3 gene fusion-related RCC (Xp11.2 translocation RCC), a rare type of tumor. Although its overall incidence rate is very low, about 1 / 3 of children with RCC is this type of RCC, and retrospective studies have shown that such tumors are not uncommon in my country. The applicant of this project used immunohistochemical method to diagnose and report 11 cases of this tumor in China for the first time in 2007. The latest research shows that the prognosis of RCC with Xp11.2 translocation is worse than that of RCC with non-Xp11.2 translocation. And the prognosis ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 饶秋周晓军马恒辉王璇刘标
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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