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Kit for detecting gene variation in tumor FFPE (formalin-fixed paraffin-embedded) sample

A gene mutation and tumor-related gene technology is applied in the field of gene mutation kits in tumor FFPE samples, which can solve the problems of inaccurate detection and limitations of the applicability of multiplex PCR methods.

Pending Publication Date: 2017-05-31
ANNOROAD GENE TECH BEIJING +2
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  • Claims
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Problems solved by technology

[0005] At present, many companies have introduced kits for enriching tumor-related gene regions by multiplex PCR methods, such as Qiagen’s GeneRead DNAseq Colorectal Cancer Panel, Illumina’s TruSight Tumor 15, Illumina’s TruSeq Amplicon Cancer Panel, etc., but these kits only It can detect SNV. For CNV and FUSION, this kind of products cannot be accurately detected. Therefore, the applicability of the multiplex PCR method is greatly limited

Method used

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  • Kit for detecting gene variation in tumor FFPE (formalin-fixed paraffin-embedded) sample
  • Kit for detecting gene variation in tumor FFPE (formalin-fixed paraffin-embedded) sample
  • Kit for detecting gene variation in tumor FFPE (formalin-fixed paraffin-embedded) sample

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Embodiment 1

[0053] 1.1 Design and synthesis of liquid chip for tumor gene detection

[0054] According to literature reports and information from multiple databases (NCBI-PDB, PubMed, COSMIC, Clinvar, HGMD, GeneCards, zj-LOVD, etc.), target genes or specific target regions for blood disease detection were selected for probe design. The designed probes are compared with NCBI and other databases by Blast, and it is necessary to ensure that the number of bases of the probes is between 60 and 80 nt, the TM value is moderate, there is no palindromic sequence and other special structures, and the sequence homology is 50% Between ~80%, these values ​​were adopted to ensure the specificity of the probe.

[0055] The designed probe sequences were synthesized into oligonucleotide probes, and after PCR verification, a tumor-associated gene capture probe clone library was obtained.

[0056] Amplification primers or linker elements with a certain number and position of biotin labels are used to prepa...

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Abstract

The invention provides a kit for detecting gene variation in a tumor FFPE (formalin-fixed paraffin-embedded) sample. The kit comprises probes for gene trapping, and the probes for gene trapping include a probe designed for an SNV (single nucleotide variant) area, a probe designed for a CNV (copy number variation) area, and a probe designed for a FUSION area; public primers include nucleotide sequence primers shown in SEQ ID NO:1; and tag primers include nucleotide sequence primers shown in SEQ ID NO:2-SEQ ID NO:5. According to the invention, in a method for detecting gene variation in the tumor FFPE sample, the genes SNV, CNV and FUSION can be stably detected at the same time.

Description

technical field [0001] The invention belongs to the field of molecular in vitro diagnosis, and in particular relates to a kit especially suitable for gene variation in tumor FFPE samples. Background technique [0002] Tumor tissues are mostly preserved in the form of FFPE (formalin-fixed paraffin-embedded, formalin-fixed paraffin-embedded) samples. The huge number of archived FFPE samples provides a valuable resource for retrospective studies, elucidation of disease mechanisms, discovery of therapeutic targets and indication of prognosis. However, the research on FFPE samples is extremely challenging. First of all, paraffin-embedded medical samples are very precious, and the total amount of each sample is very limited and irreplaceable. Secondly, during the manufacturing process of FFPE samples, the fixation of formalin will cause the nucleic acid in the tissue to be degraded to varying degrees and cross-linked between molecules. Nucleic acids in DNA also have a huge impac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6806C12Q1/6869C12Q1/6886C12Q2600/156C12Q2600/166C12Q2535/122C12Q2565/519C12Q2531/113
Inventor 王旺荆瑞琳董永芳玄兆伶李大为梁峻彬陈重建
Owner ANNOROAD GENE TECH BEIJING
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