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Fusion nucleic acid, fusion protein, and kit for detecting secondary glioblastoma

一种胶质母细胞瘤、融合蛋白的技术，应用在检测继发胶质母细胞瘤的试剂盒领域，能够解决继发性胶质母细胞瘤和原发性胶质母细胞瘤准确性有待提高等问题，达到提高准确性的效果

Active Publication Date: 2016-01-20
BEIJING PEARL BIOTECH LIMITED LIABILITY CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Therefore, the accuracy of existing detection techniques in distinguishing secondary glioblastoma from primary glioblastoma still needs to be improved

Method used

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  • Fusion nucleic acid, fusion protein, and kit for detecting secondary glioblastoma
  • Fusion nucleic acid, fusion protein, and kit for detecting secondary glioblastoma
  • Fusion nucleic acid, fusion protein, and kit for detecting secondary glioblastoma

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0051] In this preparation example, secondary glioblastoma samples and primary glioblastoma samples were obtained, and their RNA and cDNA were obtained.

[0052] Table 1

[0053]

[0054] 59 samples of primary glioblastoma (primary glioblastoma) and 20 samples of secondary glioblastoma (secondary glioblastoma) were collected using the operation in accordance with the standards of the medical ethics committee. Among them, each patient who collects samples has obtained the consent of himself and his therapist before collecting samples, and has written certification materials. Among them, the diagnosis and differentiation of primary glioblastoma and secondary glioblastoma are based on the histology in the literature (LouisDN, et al, 2007. The2007WHOclassificationoftumoursthecentralnervoussystem.ActaNeuropathol114(2):97-109.) method carried out. The gender, age, and disease type information of the pathological samples are shown in Table 1, where pGBM represents primary gliobl...

Embodiment 1

[0057] In this embodiment, the RNA of 59 primary glioblastoma samples and 20 secondary glioblastoma samples collected in Preparation Example 1 were sequenced.

[0058] The RNA of each sample was constructed into an RNA library using an RNA library construction kit (purchased from Illumina), and then RNA sequencing was performed on the RNA library using a sequencing platform (Illumina HiSeq2000). The sequence obtained by sequencing was compared with the reference RNA sequence database (Hg19Refseq, GRCh37), and the method in the reference (McPhersonA, etal.

[0059] As a result, it was found that in the samples shown in Table 1, the RNA of the fusion gene of the present invention appeared in the samples of multiple secondary glioblastomas (sGBM), but the primary glioblastoma (pGBM) No RNA of the fusion gene of the present invention appeared in any of the samples, and the specific occurrences are shown in Table 2.

[0060] Table 2

[0061]

[0062] It can be seen from the da...

Embodiment 2

[0065] In this embodiment, PCR verification of the fusion protein was carried out on the cDNA obtained from RNA prepared from 59 samples of primary glioblastoma and 20 samples of secondary glioblastoma collected in Example 1.

[0066] The primers used for PCR verification consist of the first primer shown in SEQ ID NO:17 and the second primer shown in SEQ ID NO:18. The operation of PCR was carried out according to the instructions of synthetic primers and PCR kit. The product of PCR displays the presence or absence of amplified bands through agarose gel nucleic acid electrophoresis, and the amplified bands that appear are recovered using a DNA gel recovery kit (QIAquickPCR purification kit, purchased from Qiagen), and then cloned into a T vector (pGEM - Teasyvector, purchased from Promega), and sequenced with a DNA sequencer (ABIPrism3730×1 DNA Sequencer, purchased from Applied Biosystems). The results are shown in Table 3

[0067] table 3

[0068]

[0069] It can be see...

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Abstract

The present invention provides a fusion protein specifically appearing in secondary glioblastoma, and a fusion nucleic acid encoding the fusion protein. The present invention further provides a kit for detecting secondary glioblastoma. With the technical scheme, the accuracy of the distinguishing of the secondary glioblastoma and the primary glioblastoma is effectively improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a fusion protein, a fusion nucleic acid encoding the fusion protein and a kit for detecting secondary glioblastoma. Background technique [0002] Glioblastoma is the most malignant glioma among astrocytic tumors. The tumor is subcortical and most often grows anywhere in the supratentorial hemisphere. It grows infiltratively and often invades several lobes and deep structures. It can also spread to the contralateral cerebral hemisphere through the corpus callosum. The most common site of occurrence is the frontal lobe, followed by the temporal lobe and parietal lobe, and a few can be found in the occipital lobe / thalamus and basal ganglia. [0003] Glioblastoma grows fast and has a short course of disease. 70-80% of patients have a course of 3-6 months, and only 10% of patients have a course of more than 1 year. In individual cases, due to tumor hemorrhage, stroke-like onset may occ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/11C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886C12N9/12C12N9/16C12Q2600/158C12Y207/10001C12Y301/03048G01N33/57407G01N2333/912G01N2333/916
Inventor 江涛保肇实
Owner BEIJING PEARL BIOTECH LIMITED LIABILITY CO
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