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Application of miRNA (Micro Ribonucleic Acid) 222 to prostatic cancer serological diagnosis kit

A technology of mir-222 and rnamir-222, 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) 222 to prostatic cancer serological diagnosis kit
  • Application of miRNA (Micro Ribonucleic Acid) 222 to prostatic cancer serological diagnosis kit
  • Application of miRNA (Micro Ribonucleic Acid) 222 to prostatic cancer serological diagnosis kit

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Experimental program
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Effect test

Embodiment 1

[0031] The present invention firstly provides an application of a reagent for detecting microRNA miR-222 and its precursor in a blood sample in the preparation of a test kit for diagnosing and selecting a treatment plan for prostate cancer. 222 (as shown in SEQ ID NO: 1) and its precursor specific reagents, specifically: PCR primers and probes specific for miR-222 and its precursor detection and the gene prepared using the probe chip.

[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-222 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-222-specific primers; in a preferred example, the miR-222-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-222 shown as SEQ ID NO: 1) to a prostatic cancer serological diagnosis kit, and in particular relates to application of a reagent for detecting miR-222 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-222 and a precursor thereof, and is concretely a probe and a PCR (Polymerase Chain Reaction) primer for detecting the specificity of the microRNAmiR-222 and the precursor thereof. The invention further relates to a prostatic cancer diagnosis kit comprising a polymerase chain reaction reagent, a miR-222 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-222 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) 222 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 subtypes ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王玉琢石建党张琚成飒杨阔杜小玲原辉徐勇罗飞
Owner NANKAI UNIV