Novel technology for diagnosing liver cancer by utilizing microRNAs in serum

A technology for micro-ribonucleic acid and liver cancer, which can be applied to nucleotide libraries, microbial determination/inspection, chemical libraries, etc., can solve the problems of time-consuming, labor-intensive, difficult, and rough, and achieve the effect of simple and clear results and low cost.

Inactive Publication Date: 2011-03-23
JIANGSU MICROMEDMARK BIOTECH
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Problems solved by technology

[0006] At present, the biochemical and molecular biological methods for clinical diagnosis of diseases are relatively cumbersome and rough. For example, for the diagnosis of cancer, it is necessary to perform histocytological staining on surgical specimens. The process is complicated and time-consuming.
In addition, there is currently no set of methods

Method used

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  • Novel technology for diagnosing liver cancer by utilizing microRNAs in serum
  • Novel technology for diagnosing liver cancer by utilizing microRNAs in serum
  • Novel technology for diagnosing liver cancer by utilizing microRNAs in serum

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Embodiment Construction

[0013] 1. RT-PCR of microRNA in serum

[0014] First of all, it must be determined that microRNAs can be detected in serum. We have used RT-PCR technology to prove that not only microRNAs can be detected in serum, but also their expression is quite abundant. The specific steps are: collect the serum of normal people and certain patients; there are two schemes for subsequent operations, one is to use 10 μl of serum as a buffer for direct reverse transcription, and the other is to extract total RNA from serum using TRIzol Reagent (invitrogen), 10ml serum can usually enrich about 10μg of RNA; after reverse transcription, add 4μl 5×AMV buffer, 2μl 10mM each dNTP mixture (Takara), 0.5μl RNase Inhibitor (Takara), 2μl AMV (Takara) And 1.5μl gene-specific reverse primer mixture, incubate the mixture at 16°C for 15min, react at 42°C for 60min for reverse transcription, and incubate at 85°C for 5min to inactivate AMV enzymes; finally for PCR and electrophoresis: cDNA by 1 / 50 dilution,...

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Abstract

The invention provides the combination of microRNAs used for evaluating the liver cancer condition of a subject and contains microRNAs which exist stably and are detectable in the serum/plasma of the subject. A method for evaluating the liver cancer condition of the subject comprises a step of measuring the microRNAs which exist stably and are detectable in the serum/plasma of the subject. A kit used for evaluating the liver cancer condition of the subject contains a tool for measuring the microRNAs which exist stably and are detectable in the serum/plasma of the subject. A biochip for evaluating the liver cancer condition of the subject contains a component for measuring the microRNAs which exist stably and are detectable in the serum/plasma of the subject. The novel technology can be used in the aspects of the diagnosis and the differential diagnosis, the prognosis of the generation and the recurrence of the disease complication and the therapeutic effect evaluation of liver cancer, the screening and the pharmacological effect evaluation of medicinal active ingredients and the like.

Description

1. Technical field [0001] The invention belongs to the field of biotechnology, and relates to the separation, qualitative and quantitative analysis of human serum microribonucleic acid molecules. It also involves various clinical indications of liver cancer. Specifically, it is a new technology that uses the special changes of microRNA in serum to diagnose liver cancer, predict the pathogenesis of liver cancer, judge the occurrence of complications, the probability of recurrence, and the prognosis of liver cancer, and analyze the efficacy and efficacy of drugs. 2. Technical Background [0002] MicroRNA, called microRNA in English, is a class of non-coding single-stranded small RNA molecules with a length of about 19 to 23 nucleotides. They are highly conserved in evolution and widely exist in animal and plant cells. Hundreds of microRNAs have been identified in a variety of biological species, including mice, mice, and rats. [0003] MicroRNA plays an important role beyond...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
CPCC12Q1/6886C12Q2600/178C12Q2600/158
Inventor 张辰宇曾科张俊峰李海进
Owner JIANGSU MICROMEDMARK BIOTECH
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