Exosome-associated microRNA as a diagnostic marker

An exosome, labeling technology, applied to the determination/inspection of microorganisms, coin-operated equipment for distributing discrete items, biological testing, etc.

Active Publication Date: 2010-06-23
UNIV OF LOUISVILLE RES FOUND INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The greatest difficulty in most current mass spectrometry methods is dynamic range, not sensitivity

Method used

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  • Exosome-associated microRNA as a diagnostic marker
  • Exosome-associated microRNA as a diagnostic marker
  • Exosome-associated microRNA as a diagnostic marker

Examples

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preparation example Construction

[0092] Biopolymer arrays (such as polynucleotide microarrays) can be manufactured by depositing previously obtained biopolymers (for example, from synthetic or natural sources) on a substrate or by in-situ synthesis methods. Methods of depositing to obtain biopolymers include, but are not limited to: loading a needle or capillary, and then contacting a surface, such as described in US Patent No. 5,807,522; deposition by ejecting from a pulse jet nozzle such as an ink jet head, such as PCT Publication WO95 / 25116 and WO 98 / 41531 and elsewhere. In-situ manufacturing methods include the following methods and pulse jetting can also be used to deposit reagents: those methods described in US Patent 5,449,754 for synthesizing peptide arrays, and US Patent 6,180,351, WO 98 / 41531 and references cited herein are used for Those methods for polynucleotides. The details of precipitating previously obtained biopolymers or manufacturing biopolymer arrays by in-situ methods are further descri...

Embodiment 1

[0115] EPCAM-positive circulatory exosomes are present in women with benign and malignant ovarian diseases

[0116] Use anti-EpCAM antibody magnetic beads to specifically isolate EpCAM-positive exosomes, measure the total protein of these circulatory exosomes, and map them relative to the stage of the disease ( image 3 A). The level of EpCAM-positive exosomes in age-matched normal volunteers (controls) was 0.039±0.030 mg / ml exosomal protein, which represents the background value of the assay. Patients diagnosed with benign ovarian disease have 0.149±0.065 mg / ml exosomal protein, which is significantly higher than the control value. Patients diagnosed with ovarian cancer all showed significantly high levels of EpCAM-positive exosomes (compared to benign diseases or controls). Women suffering from stage I ovarian cancer presented 0.320±0.056 mg / ml of circulatory system exosomal protein, which was significantly higher than that of controls and benign diseases (p image 3 B). Throu...

Embodiment 2

[0118] Association of small RNA with tumor-derived exosomes

[0119] In order to identify whether these isolated exosomes contain small RNAs, check them using Bio-Analyzer 2100 ( Figure 4 ). These analyses identified the presence of a significant number of small RNAs in the absence of 18S and 28S RNAs that normally occur with cell-derived RNA. These materials are then used for miRNA profiling.

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Abstract

The presently disclosed subject matter provides methods of diagnosis of cancer or adverse pregnancy outcomes in a subject by measuring amounts of one or more microRNAs present in cancer-derived exosomes isolated from a biological sample from the subject.

Description

[0001] Related application [0002] This application claims the priority basis based on the U.S. provisional application with serial number 60 / 951,812 filed on July 25, 2007 and serial number 61 / 050,438 filed on May 5, 2008, and the entire disclosure of these applications is by reference The way is included in this article. Technical field [0003] The subject matter disclosed in this application relates to methods for the diagnosis and prognosis of cancer and adverse pregnancy outcomes. Specifically, the subject matter disclosed in the present application relates to a diagnostic and prognostic method that is based on determining in a biological sample from a subject one or more types of foreign sources related to cancer or adverse pregnancy outcomes. The amount of microRNA in the exosome. Background technique [0004] The identification of cancer biomarkers suitable for the early detection and diagnosis of cancer provides great hope for improving the clinical outcome of subjects....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCC12Q1/6809C12Q2600/112C12Q1/6886C12Q2600/178Y10T436/143333G07F17/3211G07F17/3258C12Q2563/161C12Q2525/207C12Q1/6813C12Q1/6883G01N33/574G01N33/57423G01N33/57449G01N33/57492C12Q2600/118C12Q2600/158
Inventor D·D·泰勒C·格塞欧-泰勒
Owner UNIV OF LOUISVILLE RES FOUND INC
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