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A matr3-tfe3 fusion gene and detection primer and application thereof
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A MATR3-TFE3, fusion gene technology, applied in the field of medical testing, can solve the problems of long testing period, scarce testing platform, and high sample quality requirements, achieve reliable detection rate, improve detection rate and accuracy, and expand detection range. Effect
Active Publication Date: 2018-03-09
NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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[0006] At present, high-throughput sequencing is the only detection method that can clarify unknown translocation sites. However, high-throughput sequencing is expensive, the detection cycle is long, the detection platform is scarce, and the requirements for sample quality are high, which is not conducive to popularization and promotion. For most patients It is not the preferred detection method
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Embodiment 1
[0026] Embodiment 1 is verified for the case of definite diagnosis:
[0027] For cases in which new fusion sites of MATR3 exon15-TFE3 exon4 were detected by high-throughput sequencing RNA-seq, the primers we designed were used for verification.
[0028] 1. Extraction of RNA:
[0029] Strictly follow the RNeasy FFPE Kit operating instructions for extraction. ① Dewaxing: dewax the collected slides with xylene, rinse with absolute ethanol, air-dry and scrape off with a scalpel blade and put them into a 1.5ml EP tube; Proteinase K, mix well, enzymatically digest at 56°C for 15min, then at 80°C for 15min, cool on ice; ③add 16μl DNase buffer, then add 10μl DNase I, mix well, let stand at room temperature for 15min, centrifuge at 12000rpm for 15min, and take the supernatant;④ Add 320μl of binding solution and 720μl of absolute ethanol, mix well, transfer to the adsorption column twice, centrifuge at 8000rpm for 1min, discard the waste liquid; ⑤Wash: add 500μl of washing solution, c...
Embodiment 2
[0033] Example 2 Detection of Xp11.2 tumors with unknown translocation partners
[0034] We used the primer combination of the present invention to detect 35 Xp11.2 tumors with unknown translocation partners, and the RNA extraction, reverse transcription PCR and sequencing methods were the same as above.
[0035] Result: using the primer combination detection designed by the present invention, a total of 1 case detected the MATR3-TFE3 fusion gene ( image 3 , Figure 4 ), verified by immunohistochemistry, fluorescence in situ hybridization and other methods, the results are true and reliable.
[0036] Evaluation: The primer combination of the present invention is a supplement to the original Xp11.2 fusion gene primers, covers the unreported sites of the TFF3 fusion gene, and increases the detection rate of the RT-PCR method for diagnosing the tumor.
[0037] Nanjing General Hospital of Nanjing Military Region of the Chinese People's Liberation Army
[0038] A MATR3-TFE3 f...
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Abstract
The invention discloses a novel Xp11.2 translocation partner, a detection primer and the application of the detection primer. The novel Xp11.2 translocation partner is translocation of a MATR3-TFE3 gene, the gene is located on a No.5 chromosome 5q31.2, and the gene fusion occurs between a No.15 exon of MATR3 and a No.4 exon of TFE3. The PCR primer used for detecting a MATR3-TFE3 translocation tumor is used for detecting translocation of the MATR3-TFE3 gene. The invention also discloses the application of the PCR primer to preparation of a diagnostic reagent for the MATR3-TFE3 translocation tumor. According to the novel fusion locus discovered in the high-throughput sequencing, the specific PCR primer is designed, the detection range of the original primer combination is expanded, the PCR primer can be clinically applied, and the detection rate and the accuracy in the diagnosis of the tumor are improved.
Description
technical field [0001] The invention belongs to the field of medical examination, and relates to a MATR3-TFE3 fusion gene, detection primers and applications thereof. 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). The TFE3 gene is located at XP11.2, and it is the only driving gene of Xp11.2 translocation RCC. The fusion gene can highly express the TFE3 fusion protein through the transformation of the promoter, and TFE3, as a transcription factor, binds to a specific DNA structure, transcriptionally regulates the expression of various genes in the body, and eventually causes disease. At least 10 different translocation partners and fusion genes have been reported, including ASPL-TFE3, PRCC-TFE3, SFPQ-TFE3, NONO-TFE3, CLTC-TFE3, LUC7L3-TFE3, KHSRP-TFE3, PARP14-TFE3, DVL2 -TFE3 and RBM10-TFE3, etc., there is only a single translocation ...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62
CPCC12Q1/6886C12Q2600/112
Inventor 夏秋媛饶秋叶胜兵李芳秋周晓军
Owner NANJING GENERAL HOSPITAL NANJING MILLITARY COMMAND P L A
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