Biomarker detection method and detection kit

A biomarker and detection method technology, applied in the field of biomarker detection methods and detection kits, can solve the problems of high cost, complicated operation, and single detection target, and achieve the goal of simple method and high specificity fluorescence detection Effect

Pending Publication Date: 2021-09-07
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods have good sensitivity and specificity, they still have the disadvantages of single detection target, high cost, long time-consuming and complicated operation.

Method used

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  • Biomarker detection method and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] In this embodiment, the target miRNA is miR-21, and its sequence is: 5'-UAGCUUAUCAGACUGAUGUUGA-3';

[0069] The sequence of the TP chain is:

[0070] 5'-CAGTCTGATAAGCTAACCCTAACCCTAACCCTAAAATCCGTCGAGCAGAGTT-3';

[0071] The sequence of the auxiliary chain AS is: 5'-CTCGAGATAAGGTTAGCTTA-3'.

[0072] The sequence of the hairpin DNA probe H1 is:

[0073] 5'-ACGTTAAACCTTATCTCGAGTTCGACGAATAAGGTT-3';

[0074] The sequence of the hairpin DNA probe H2 is:

[0075] 5'-ATAAGGTTTAACGTAACCTTATTCGTCTTAAAAACCCCCTAATTCCCCC-3'.

[0076] In this embodiment, the detection method of the biomarker comprises the following steps:

[0077] 1) Synthesis of DNA-templated silver nanoclusters H2-AgNCs:

[0078] Mix hairpin DNA probe H2 and silver nitrate solution in PBS (pH7.4), heat to 95°C, keep for 5 minutes, then slowly cool to room temperature, then add sodium borohydride to the reacted solution, and stir vigorously After 1 minute of reaction, H2-AgNCs were obtained and kept under dark...

Embodiment 2

[0099] In this embodiment, the target miRNA is miR-141, and its sequence is: 5'-UAACACUGUCUGGUAAAGAUGG-3';

[0100] The sequence of the TP chain is:

[0101] 5'-TACCAGACAGTGTTAACCCTAACCCTAACCCTAAAATCCGTCGAGCAGAGTT-3';

[0102] The sequence of the auxiliary chain AS is: 5'-CTCGAGATAAGGTTGGGTTAACACT-3'.

[0103] Hairpin DNA probes H1 and H2 are the same as in Example 1.

Embodiment 3

[0105] In this embodiment, the target miRNA is miR-183, and its sequence is: 5'-UAUGGCACUGGUAGAAUUCACU-3';

[0106] The sequence of the TP chain is:

[0107] 5'-TCTACCAGTGCCATAACCCTAACCCTAACCCTAAAATCCGTCGAGCAGAGTT-3';

[0108] The sequence of the auxiliary chain AS is:

[0109] 5'-CTCGAGATAAGGTTGGGTTATGGCACT-3'.

[0110] Hairpin DNA probes H1 and H2 are the same as in Example 1.

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Abstract

The invention discloses a biomarker detection method and a detection kit. The biomarker is any one or two of target miRNA and telomerase; and the method comprises the following steps of mixing a sample to be detected, a DNA templated silver nanocluster mixture H2-AgNCs@GO subjected to fluorescence quenching by graphene oxide, an auxiliary chain AS, a TP chain and a hairpin DNA probe H1 for reaction; and calculating the concentration of the biomarker in the sample to be detected by detecting the fluorescence intensity of a reaction product. The DNA template silver nanocluster is used as a fluorescence signal, and the signal amplification performance of fluorescence resonance energy transfer and hybridization chain reaction is combined, so that high-sensitivity and high-specificity fluorescence detection of miRNA and telomerase is realized.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a detection method and a detection kit of a biomarker. Background technique [0002] MicroRNA (miRNA) and telomerase are important biomarkers with high medical value, especially the joint detection of the two biomarkers has good application value. At present, the widely reported detection methods for telomerase activity and miRNA analysis mainly include polymerase chain reaction (PCR)-based classical telomeric repeat amplification (TRAP) and nanomaterial-based sensing platforms, etc. Although these methods have good sensitivity and specificity, there are still shortcomings such as single detection target, high cost, long time-consuming, and complicated operation. Therefore, it is necessary to develop a fast, simple protocol capable of detecting multiple targets simultaneously. Contents of the invention [0003] The technical problem to be solved by the present invention is...

Claims

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Application Information

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IPC IPC(8): C12Q1/6818C12Q1/48
CPCC12Q1/6818C12Q1/48C12Q2525/207C12Q2525/301C12Q2563/107C12Q2521/113C12Q2565/101C12Q2537/1373C12Q2563/137
Inventor郭振振陈锡峰蒋依婷
OwnerSUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI