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Tumor marker miRNA detection probe and detection chip

A tumor marker and detection chip technology, which is applied in the detection chip and preparation field of tumor marker protein and nucleic acid, can solve the problems of low abundance, small size, and limited effective application, and achieve low detection limit and high detection accuracy Effect

Active Publication Date: 2021-06-01
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, miRNA has the characteristics of small size (19-24 bases), low abundance, sequence homology and easy degradation, so it is difficult to achieve quantitative detection by traditional inspection methods
[0004] The current quantitative detection of miRNA includes Northern blotting, qRT-PCR, fluorescence method, microarray method, and SERS method, but most of them are limited to single microRNA detection, resulting in no reference for microRNA detection
Real-time quantitative polymerase chain reaction (qRT-PCR) is widely used in nucleic acid detection, but due to the short chain length of miRNA, it is difficult to design primers, which increases the complexity of the experimental process
In addition, qRT-PCR cannot measure the absolute content of miRNA in serum, only the relative content can be given
Due to the lack of a unified miRNA internal reference, it is difficult to compare the results of different samples, which also limits its effective clinical application

Method used

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  • Tumor marker miRNA detection probe and detection chip
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  • Tumor marker miRNA detection probe and detection chip

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] For the preparation method of tumor marker miRNA detection probe, see figure 1 , including the following steps:

[0059] (1) Three kinds of DNA single strands were synthesized respectively ( figure 1 1, 2 and 3) in which one DNA single strand is composed of a total of 78 nucleotides in the first part ① and the second part ②, and the free end of the second part is connected with a linker molecule 4(-(CH 2 ) 3 -NH 3 ); the other two DNA single strands (indicated by 2 and 3 respectively) consist of a total of 102 nucleotides in the first part, the second part and the third part; the third part ③ is the cohesive end of the 24 nucleotide sequence The number of nucleotides in the first part and the second part of the three DNA single strands is equal, and the first part in each DNA single strand is reversely complementary to the nucleotide sequence of the second part of the adjacent DNA single strand; The cohesive ends can be reverse-complementary paired with the nucleo...

Embodiment 2

[0068] For the preparation method of tumor marker miRNA detection probe, see figure 2 , including the following steps:

[0069] (1) Synthesize 7 kinds of DNA single strands respectively, one of which is composed of the first part and the second part with a total of 20 nucleotides, and the free end of the second part is connected with a linking molecule (-(CH 2 ) 12 -NH3); the remaining 6 DNA single strands consist of a total of 39 nucleotides in the first part, the second part and the third part; the third part is the cohesive end of the 19 nucleotide sequence; of the 7 DNA single strands The number of nucleotides in the first part and the second part is equal, and in each DNA single strand, the first part is reversely complementary to the nucleotide sequence of the second part of the adjacent DNA single strand; The nucleotide sequences of the detected miRNAs are paired in reverse complementarity.

[0070] The nucleotide sequence of one DNA single strand is shown in SEQ ID...

Embodiment 3

[0082] A method for preparing a tumor marker miRNA detection probe, comprising the steps of:

[0083] (1) Three kinds of DNA single strands were synthesized respectively, one of which consisted of a total of 28 nucleotides in the first part and the second part, and a linker molecule (-(CH 2 ) 6 -NH 3 ); the other two DNA single strands consist of a total of 50 nucleotides in the first part, the second part and the third part; the third part is the cohesive end of 22 nucleotide sequences; the first part of the three DNA single strands The number of nucleotides in the second part is equal, and the first part of each DNA single strand is reversely complementary to the nucleotide sequence of the second part of the adjacent DNA single strand; The nucleotide sequences of miRNAs are paired in reverse complementarity.

[0084] The nucleotide sequence of one DNA single strand is shown in SEQ ID NO.11. (28)

[0085] SEQ ID NO.11: GAAGCTGCCAGTAC

[0086] The other two DNA singl...

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Abstract

The invention discloses a tumor marker miRNA detection probe and a detection chip. The tumor marker miRNA detection chip includes a substrate loaded with nano-metal, and the substrate loaded with nano-metal is connected with 2-3 kinds of signal molecules, and the signal Molecules belong to different classes of active carboxyl signal molecules, carboxyl signal molecules or amino signal molecules classified by containing chemical groups; any one of the 2-3 kinds of signal molecules is connected to the antibody, and the remaining 1-2 kinds are connected by the connecting molecule Claim 1. Tumor marker miRNA detection probe. The tumor marker miRNA detection chip of the present invention can realize simultaneous quantitative and non-interference detection of different types of marker proteins and nucleic acids; using nano-stress sensing, the concentration information of different types of marker proteins and nucleic acids can be converted into characteristic peak displacements of signal molecules , so as to detect the absolute concentration of different types of marker proteins and nucleic acids, and the detection accuracy is higher.

Description

technical field [0001] The invention belongs to a detection chip for tumor marker protein and nucleic acid and a preparation method thereof, in particular to a quantitative and highly sensitive simultaneous detection probe and detection chip for tumor marker protein and nucleic acid. Background technique [0002] Liver cancer is one of the deadliest malignant tumors in the world, with a mortality-to-morbidity ratio greater than 95%. Liver cancer is a cancer with late onset of clinical symptoms. Surgical resection or liver transplantation is the only effective treatment so far. Therefore, early diagnosis of liver cancer is the key to reducing the ratio of mortality to morbidity. [0003] A typical marker of liver cancer is alpha-fetoprotein (AFP), but it shows low sensitivity and specificity in the early stage of disease diagnosis. There are still 30%-40% of liver cancer patients whose AFP test is negative, including patients with intrahepatic cholangiocarcinoma (ICC), well-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574
CPCG01N33/57484
Inventor 李敏仰大勇张杰程林秀
Owner TIANJIN UNIV