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Liver cancer diagnostic marker composed of blood plasma microRNA (micro ribonucleic acid) and new method for diagnosing liver cancer

A diagnostic marker, hsa-mir-21 technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of limited number of microRNA, lack of validation, small sample size, etc.

Active Publication Date: 2013-07-10
JUSBIO SCI SHANGHAI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above results suggest that the diagnosis of HCC by serum microRNA is feasible, but there are many defects in these studies, such as the limited number of microRNA used for screening, small sample size or lack of independent verification

Method used

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  • Liver cancer diagnostic marker composed of blood plasma microRNA (micro ribonucleic acid) and new method for diagnosing liver cancer
  • Liver cancer diagnostic marker composed of blood plasma microRNA (micro ribonucleic acid) and new method for diagnosing liver cancer
  • Liver cancer diagnostic marker composed of blood plasma microRNA (micro ribonucleic acid) and new method for diagnosing liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0165] Example 1: Collection and preparation of plasma samples:

[0166] The experiments in this invention have been approved by the local ethics committee and informed consent was obtained from all patients. The experimental design of the screening stage, training stage and verification stage of microRNA biomarkers in the present invention is as follows: figure 1 shown. The main method steps for identifying plasma samples from HCC patients with the recommended plasma microRNA panels are as follows: figure 2 shown.

[0167] Between August 2008 and June 2010, 934 met the eligibility criteria ( figure 2 ) blood samples were collected in advance from Shanghai Zhongshan Hospital and Public Health Center. These samples were obtained from 167 healthy donors (healthy group), 169 patients with chronic hepatitis B (CHB group), 141 patients with cirrhosis after HBV infection (cirrhosis group), and 457 HCC patients associated with HBV infection (HCC group) . These samples were di...

Embodiment 2

[0182] Embodiment 2: microRNA microarray analysis:

[0183] Qualitative analysis of microRNA in specific plasma samples was performed using the Agilent microRNA microarray platform (Agilent microRNA microarray platform, Agilent Technologies, Santa Clara, CA, USA). The microarray contains probes for 723 human microRNAs from Sanger database v.10.1. Total RNA (100 ng) from any one of the 137 plasma samples was spiked into single-color CY3 for labeling. Scan the gene chip with XDR scanner (PMT100, PMT5), and perform labeling and hybridization according to the protocol of Agilent microRNA microarray system. Microarray image information was converted into spot intensity values ​​by Feature Extraction Software Rev. 9.5.3 (Agilent Technologies, Santa Clara, CA). The signal after background removal was normalized with the stable internal reference hsa-miR-1228. Then, a base-2 log transformation is performed. An unreliable sample with an inter-slice variation coefficient (CV) of mor...

Embodiment 3

[0186] Example 3: Microarray data of 137 samples

[0187] The expressions of the 15 candidate microRNAs used for further verification in the microarray analysis are shown in Table 5, and the microRNAs meeting the selection criteria are indicated in bold.

[0188] table 5

[0189] Expression of 15 candidate microRNAs in microarray

[0190]

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Abstract

The invention relates to a liver cancer diagnostic marker composed of blood plasma microRNA (micro ribonucleic acid), a kit containing the marker and a new method for diagnosing liver cancer (in particular early liver cancer). The liver cancer diagnostic marker is composed of a plurality of nucleic acid molecules, wherein each nucleic acid molecule is used for coding at least one microRNA sequence; and preferably, the liver cancer diagnostic marker is composed of nucleic acid molecules for coding hsa-miR-122, hsa-miR-192, hsa-miR-21, hsa-miR-223, hsa-miR-26a, hsa-miR-27a, hsa-miR-801 and hsa-miR-1228. The kit can be applied to diagnosing hepatocellular carcinoma, in particular early hepatocellular carcinoma, or to differentiating the blood plasma of at least one hepatocellular carcinoma sufferer from the blood plasma of at least one healthy individual, and the blood plasma of at least one chronic hepatitis b sufferer or the blood plasma of at least one cirrhosis sufferer.

Description

technical field [0001] The invention relates to a liver cancer diagnostic marker composed of plasma microRNA and a new method for diagnosing liver cancer (especially early liver cancer). The plasma microRNA includes hsa-miR-122, hsa-miR-192, hsa-miR-192, hsa- miR-21, hsa-miR-223, hsa-miR-26a, hsa-miR-27a and hsa-miR-801. Background technique [0002] Hepatocellular carcinoma (HCC) is one of the most common solid malignant tumors worldwide. It represents the major histological type of liver cancer and may account for 70%-85% of all liver cancer cases. Approximately 600,000 new cases and almost the same number of deaths occur worldwide each year, reflecting the lack of effective early diagnosis and treatment of the disease (Thorgeirsson, S.S. and Grisham, J.W. (2002) Nat Genet 31, 339-346 ; Jemal A et al. (2011) CA Cancer J Clin 61:69-90.; Bosch F.X. et al (2004) Gastroenterology 127, S5-S16; Perz J.F. et al. (2006) J Hepatol 45, 529-538) . [0003] Hepatocellular carcinom...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12Q1/68
Inventor 樊嘉周俭代智於雷胡捷王征
Owner JUSBIO SCI SHANGHAI CO LTD
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