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Detection card for rapidly detecting micro ribonucleic acid (miRNA) as well as preparation method and application of detection card

A microRNA and detection card technology, applied in the field of detection, can solve the problems of inability to meet clinical needs, poor repeatability, long time consumption, etc., and achieve the effect of improving detection sensitivity, evaluating curative effect and prognosis, and high efficiency

Inactive Publication Date: 2015-02-25
NANJING BONA TIANYUAN BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current detection of miRNA must rely on PCR amplification, which takes a long time and has poor repeatability, which cannot meet clinical needs

Method used

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  • Detection card for rapidly detecting micro ribonucleic acid (miRNA) as well as preparation method and application of detection card
  • Detection card for rapidly detecting micro ribonucleic acid (miRNA) as well as preparation method and application of detection card
  • Detection card for rapidly detecting micro ribonucleic acid (miRNA) as well as preparation method and application of detection card

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Detection of miR-21 associated with the pathogenesis of lung cancer:

[0037] Firstly, the miRNA sequence complementary to miR-21 was synthesized, a carbon chain composed of 5 carbon atoms was modified at one end, and a Cy3 fluorescent group was labeled at the other end;

[0038] A set of blank control areas and sample detection areas are defined on the surface of the nitrocellulose membrane. 100 μL, 1 μmol / L miR-21 complementary miRNA solution was sprayed on the surface of the blank control area and the sample detection area by spot spraying technology, and kept at 37 °C for 1 hour. The modified nitrocellulose membrane is further cut into a suitable size by using a test strip cutter, and then assembled with a sample pad, a nanomaterial pad and an absorbent pad to form a test card.

[0039] The miR-21 solution was mixed with gold nanoparticles with a diameter of 5 nm and coated with trisodium citrate on the surface to prepare a gold nanoparticle-labeled miR-21 ...

Embodiment 2

[0043] Example 2 Simultaneous detection of miR-24, miR-25, miR-30a, miR-100 and miR-126 associated with the onset of lung cancer:

[0044] Firstly, miRNA sequences complementary to miR-24, miR-25, miR-30a, miR-100 and miR-126 were synthesized, and a carbon chain composed of 10 carbon atoms was modified at one end, and a Cy5 fluorescent group was labeled at the other end;

[0045] Five groups of blank control areas and sample detection areas were defined on the surface of the nitrocellulose membrane. Spray 150 μL, 100 nmol / L miR-24, miR-25, miR-30a, miR-100 and miR-126 complementary miRNA solutions on the corresponding blank control area and the surface of the sample detection area respectively by spot spraying technology, and place at 37 °C Hold for 1 hour. Further use the test strip cutter to cut the modified nitrocellulose membrane into a suitable size, and then package it with the sample pad, absorbent pad, kit body, etc. into a five-in-one detection kit.

[0046] The miR...

Embodiment 3

[0048] Example 3 Simultaneous detection of miR-143, miR-145, miR-152, miR-155, miR-183, miR-188, miR-189, miR-200b, miR-320 and miR-331 associated with the onset of lung cancer:

[0049] Firstly, miRNA sequences complementary to miR-143, miR-145, miR-152, miR-155, miR-183, miR-188, miR-189, miR-200b, miR-320 and miR-331 were synthesized, respectively, in One end is modified with a carbon chain composed of 20 carbon atoms, and the other end is labeled with a Cy5 fluorescent group;

[0050] Ten groups of blank control areas and sample detection areas were defined on the surface of the nitrocellulose membrane. 2mL, 10nmol / LmiR-143, miR-145, miR-152, miR-155, miR-183, miR-188, miR-189, miR-200b, miR-320 and miR-331 complementary The miRNA solution was sprayed on the corresponding blank control area and the surface of the sample detection area, and kept at 37°C for 1 hour. Further use the test strip cutter to cut the modified nitrocellulose membrane into a suitable size, and then...

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Abstract

The invention discloses a detection card for rapidly detecting micro ribonucleic acid (miRNA). The detection card comprises a sample pad, a nanomaterial pad, a nitrocellulose membrane and an absorbent pad, wherein the nitrocellulose membrane is a detection card substrate, a sample injection zone, a blank control zone and a sample detection zone are distributed on the detection card substrate, the sample injection zone is communicated with the sample detection zone, the blank control zone and the sample detection zone are modified with probes having miRNA sequences complementary to to-be-detected miRNA, one ends of the probes are connected with the detection card substrate, and the other ends of the probes carry fluorescent groups. The invention also discloses a preparation method of the detection card. The invention also discloses a kit for rapidly detecting the miRNA associated with lung cancer. The invention also discloses application of the detection card and the detection kit. The detection card has relatively high detection sensitivity on miRNA associated with lung cancer and can simultaneously detect a plurality of miRNAs without PCR, the clinical diagnostic specificity is improved and the detection card meets application requirements of screening, diagnosis of lung cancer, determination of treatment effect and follow-up visit.

Description

technical field [0001] The invention relates to a method for detecting microribonucleic acid (miRNA), in particular to a detection card for rapidly detecting microribonucleic acid, a preparation method and application thereof, and belongs to the technical field of detection. Background technique [0002] Lung cancer ranks first in the incidence and mortality of chronic tumors in my country. It has entered the late stage when imaging and morphology are found. Currently, there are no specific markers available. How to develop a rapid diagnosis and suitable for early clinical detection of lung cancer Technology has always been a technical difficulty in this field. [0003] In recent years, with the continuous deepening of human genome research, it has gradually revealed that there is a strong correlation between MicroRNA (miRNA) in the occurrence, metastasis and apoptosis of lung cancer. miRNA is a highly conserved, non-protein-coding RNA with a length of about 21-23 nucleotide...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6816C12Q1/6886
Inventor 孔涛张寄南王炜张雷程国胜
Owner NANJING BONA TIANYUAN BIOLOGICAL TECH CO LTD
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