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SERS transverse flow test strip for nuclease detection and detection method

A flow test strip and nuclease technology, applied in the field of photochemical analysis, can solve the problems of difficult to achieve high-throughput and automatic detection, expensive instruments, complicated operation, etc., and achieve the effects of rich fingerprint information, simple operation and good selectivity.

Active Publication Date: 2020-10-09
YANTAI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional enzyme detection methods include gel electrophoresis, radioactive labeling, high performance liquid chromatography, and enzyme-linked immunosorbent assay (ELISA). Although these methods usually have high accuracy, there are still many key problems that need to be solved urgently, such as The operation is cumbersome, the cycle is long, radioactive substances need to be introduced, the equipment is expensive or the modification process is complicated, and it is difficult to achieve high-throughput and automatic detection

Method used

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  • SERS transverse flow test strip for nuclease detection and detection method
  • SERS transverse flow test strip for nuclease detection and detection method
  • SERS transverse flow test strip for nuclease detection and detection method

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Embodiment 1

[0043] Taking the detection of ExoI nuclease in the present invention as an example, ExoI nuclease can degrade the 3'-end of single-stranded DNA to produce 3'-end single-stranded nucleotides or oligonucleotides. Its preparation detection steps are as follows:

[0044] (1) Preparation of lateral flow test strips: the sample pad was soaked in 50 mM Tris-HCl containing 0.25% tritonx-100 and 150 mM NaCl, and dried in an oven at 37° C. for 2 h. The test strip used consists of three parts. The sample pad and the absorbent pad are pasted on the nitrocellulose membrane with PVC support in turn, and the contact must be ensured firmly. Cut it into strips with a width of 4 mm using a CNC cutting machine, and place the cut test strips in a sterile environment for subsequent use. Use a pipette gun to take 1 μL of anti-single-stranded DNA antibody and add dropwise to the 1 / 5 of the test strip, and drop 1 μL of 0.5 mg / mL streptavidin to the 2 / 5 of the test strip to form the control line and...

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Abstract

The invention relates to an SERS transverse flow test strip for nuclease detection and a detection method. The SERS transverse flow test strip comprises a bottom plate, a sample pad, a combination pad, a reaction film and a water absorption pad; the reaction film is provided with a detection line and a control line; the detection line is arranged at one end close to the combination pad, and the control line is arranged at one end close to the adsorption pad; the combination pad is coated with Raman reporter molecules and nanoparticles marked by detection DNA as SERS nanoprobes; streptavidin isfixed on the detection line; and an anti-single-chain DNA antibody is fixed on the control line. The SERS technology is used for quantitative analysis; the SERS signal is strong, no light drifts, thefingerprint information is rich, the stability is strong, the detection sensitivity is higher, the SERS transverse flow test strip is prepared by combining with an immunochromatography technology, the sample demand is low, the operation is simple and convenient, the reaction is fast, the result is accurate, the price is low, the detection limit is low, the selectivity is good, and the batch production is easy to realize.

Description

technical field [0001] The invention relates to a SERS lateral flow test strip for nuclease detection and a detection method, belonging to the technical field of photochemical analysis. Background technique [0002] Enzymes capable of cleaving the phosphodiester bonds of polynucleotide chains are called nucleases. Nucleases are hydrolases that act on the P-O position of the phosphodiester bond. A nuclease is a nucleic acid that acts to hydrolyze the phosphodiester bonds between nucleotides in the first step of nucleic acid breakdown. It acts on phosphodiester bonds in higher animals and plants. Nucleases from different sources have different specificities and modes of action. Some nucleases can only act on RNA, called ribonuclease (RNase), some nucleases can only act on DNA, called deoxyribonuclease (DNase), some nucleases have low specificity, can act on RNA can also act on DNA, so they are collectively called nucleases. Nucleases can be divided into exonucleases and e...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/573G01N33/558G01N33/53G01N33/52G01N21/65
CPCG01N33/587G01N33/573G01N33/558G01N33/5308G01N33/52G01N21/658
Inventor 付秀丽林浩文嘉慧付维聪
Owner YANTAI UNIV
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