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Application of miRNA (Micro Ribonucleic Acid) 423-5p to prostatic cancer serological diagnosis kit

A technology of prostate cancer, kit, applied in the field of biomedical materials

Inactive Publication Date: 2012-12-12
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although many candidate biomarkers are being discovered continuously, no new indicators and methods have been applied to clinical detection

Method used

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  • Application of miRNA (Micro Ribonucleic Acid) 423-5p to prostatic cancer serological diagnosis kit
  • Application of miRNA (Micro Ribonucleic Acid) 423-5p to prostatic cancer serological diagnosis kit
  • Application of miRNA (Micro Ribonucleic Acid) 423-5p to prostatic cancer serological diagnosis kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The present invention firstly provides an application of a reagent for detecting microRNA miR-423-5p and its precursor in a blood sample in the preparation of a kit for diagnosing and selecting a treatment plan for prostate cancer, the reagent is for microRNA miR-423-5p Reagents specific for RNAmiR-423-5p (as shown in SEQ ID NO: 1) and its precursors, specifically: PCR primers and probes specific for detecting miR-423-5p and its precursors and the use thereof The gene chip prepared by the probe.

[0032] Described specific primer is selected from the sequence shown in SEQ ID NO:2;

[0033] The specific probe is selected from the sequence shown in SEQ ID NO:5.

Embodiment 2

[0034] Embodiment 2, prostate cancer diagnostic kit

[0035] The target nucleic acid detected by the kit of the present invention is miR-423-5p having a polynucleotide sequence shown in SEQ ID NO: 1, specifically comprising:

[0036] (1) Polymerase chain reaction reagents; including hot start Taq DAN polymerase (2.5U / ul) and its reaction buffer, dNTP (10mM);

[0037] (2) miR-423-5p-specific primers; in a preferred example, the miR-423-5p-specific primers are selected from the sequence shown in SEQ ID NO:2;

[0038] (3) Nematode miR-39 reference oligonucleotide sequence: 5'-UCACCGGGUGUAAAUCAGCUUG-3' (SEQ ID NO: 3);

[0039](4) C. elegans miR-39 specific primer: 5'-CCGGGTGTAAATCAGCTTGAAA-3' (SEQ ID NO: 4);

[0040] (5) Fluorescent dye SYBR-GREEN 1;

[0041] (6) Instructions for use.

Embodiment 3

[0042] Embodiment 3, diagnosis effect experiment

[0043] 1. Using our invented xenograft model (i.e. transplanting prostate cancer tissue into the renal capsule of immunodeficient mice, the obtained tissue line is highly correlated with the pathological characteristics of clinical tumors) and next-generation sequencing technology, the metastatic and The transcriptomics of miRNAs in non-metastatic prostate cancer xenograft tissue lines (surgical specimens from the same prostate cancer patient) has been intensively studied. By comparison, we identified 104 miRNAs with differential expression, in addition to some known miRNAs and their isoforms (isomiRs), there are some new miRNAs that have not been discovered yet. Among them, 21 miRNAs have been found to be related to the occurrence of prostate cancer, and 5 miRNAs have been reported to play a role in prostate cancer metastasis, which also proves the effectiveness of our research methods from the side. In addition to those miR...

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Abstract

The invention relates to application of a miRNA (miR-423-5p shown as SEQ ID NO: 1) to a prostatic cancer serological diagnosis kit, and in particular relates to application of a reagent for detecting the miR-423-5p in a blood sample to preparation of a kit for diagnosing object prostatic cancers. The reagent has specificity to the micro RNA (Ribonucleic Acid) miR-423-5p and a precursor thereof, and is concretely a probe and a PCR (Polymerase Chain Reaction) primer for detecting the specificity of the miR-423-5p and the precursor thereof. The invention further relates to a prostatic cancer diagnosis kit comprising a polymerase chain reaction reagent, a miR-423-5p specific primer, a nematode miR-39 reference oligonucleotide sequence, a nematode miR-39 specific primer and a fluorochrome SYBR-GREEN1 which are shown in the specification. When a detection result displays that the level of the miR-423-5p or the precursor thereof in an object blood is higher than that in a reference serum, the object is prompted to suffer from the prostatic cancers. The kit can be effectively used for auxiliary diagnosis of clinical prostatic cancers.

Description

technical field [0001] The invention relates to the application of a microRNA (micro RNA, miRNA, miR) 423-5p in a prostate cancer serological diagnostic kit. It belongs to the technical field of biomedical materials. Background technique [0002] Prostate cancer (PCa) is a common malignant tumor and one of the leading causes of death among men in European and American countries. [0003] With the extension of life expectancy and the westernization of lifestyle in my country, the incidence of PCa is increasing year by year, and it is listed as one of the fastest growing tumors in China in the 21st century. Based on the 19 up-regulated miRNAs sequences screened in the previous work, this project will further detect their content in clinical serum samples to find serological biomarkers in patients with human prostate cancer metastasis. [0004] At present, the research work on the diagnostic indicators of prostate cancer mainly focuses on the following aspects: (I) PSA subtyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张琚石建党王玉琢成飒杨阔杜小玲原辉徐勇罗飞
Owner NANKAI UNIV