Aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as preparation and detection method of aptamer LC biosensor

A nucleic acid aptamer, lung surfactant technology, applied in the field of detection, to achieve the effect of rapid and efficient detection

Inactive Publication Date: 2017-12-22
JINING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the liquid crystal sensor has the advantages of fast response, no need for labeling, no need for complex and expensive reading devices, and direct detection with the naked eye, there are still few reports on the use of nucleic acid aptamers in the detection of lung surfactant protein A.

Method used

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  • Aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as preparation and detection method of aptamer LC biosensor
  • Aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as preparation and detection method of aptamer LC biosensor
  • Aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as preparation and detection method of aptamer LC biosensor

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Determination of the optimal ratio of APTES / DMOAP on the bottom slide

[0039] In order to ensure the vertical uniform orientation of the liquid crystal, and to ensure the highest possible sensitivity of the sensor, the influence of the mixing ratio of APTES / DMOAP on the orientation of the liquid crystal was investigated successively. The test results are detailed in the appendix of the manual. Figure 4 . When the volume ratio of APTES / DMOAP is 5:1, the image appears uniform black (see the appendix Figure 4 a), when the volume ratio of APTES / DMOAP is increased to 10:1, the image still presents a uniform black color (see the appendix Figure 4 b), when the volume ratio of the two continues to increase to 15:1, the Schlelen texture begins to appear in the image (see the attached manual Figure 4 c), when the volume ratio is further increased to 20:1, the image appears obvious Schlelen texture (see the appendix Figure 4 d), in order to ensure that as many nucleic aci...

Embodiment 2

[0041] Optimal concentration of aptamer (Aptamer) on the surface of slides

[0042] The amino-modified nucleic acid aptamer was immobilized on the surface of an aldylated glass slide, and it was predicted that the concentration of Aptamer would affect the orientation of the liquid crystal molecules, (see the appendix for details). Figure 5 ) SP-A Aptamer liquid crystal optical signal map) Therefore, it is necessary to further determine the appropriate concentration of Aptamer immobilized on the glass surface. When the concentration of Aptamer is high (500 nmol·L -1 , 300 nmol L -1 , 200 nmol L -1 ), due to the excessive amount of Aptamer fixed on the surface of the glass slide, it has a great influence on the orientation of liquid crystal molecules, resulting in large colored and bright areas in the image (see the appendix Figure 5 a, b, c), in order to ensure that the subsequent detection is not affected, the concentration of Aptamer needs to be reduced. When the concent...

Embodiment 3

[0044] Investigation of sensor sensitivity

[0045] Different concentrations of pulmonary surfactant protein A (SP-A) solutions were dropped on the lower glass slide immobilized with nucleic acid aptamers, and after treatment, they were assembled face-to-face with the DMOAP-modified upper glass slide to form a liquid crystal cell. Observation, the observation results are detailed in the appendix of the instruction manual Image 6 . The higher the concentration of pulmonary surfactant protein A, the more it binds to Aptamer, so the greater the damage to the vertical orientation of liquid crystal molecules, the more obvious the colored bright areas and Schlelen texture in the image, when the pulmonary surfactant protein The concentration of A is 200nmol·L -1 When the color texture optical image is seen under the polarizing microscope (see the appendix Image 6 a); when the concentration of pulmonary surfactant protein A decreased to 100 nmol L -1 , 20 nmol L -1 and 10 nmol ...

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Abstract

The invention discloses an aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as a preparation and detection method of the aptamer LC biosensor. The method is based on a detecting platform of aptamers, and when an aldehydation slide surface is modified with the aptamers of the pulmonary surfactant protein A with the optimum concentration, slight topological change on the slide surface almost has no influence on a sensing system and a dark background under a polarizing microscope is still kept; when the pulmonary surfactant protein A is present, the pulmonary surfactant protein aptamers can react with the pulmonary surfactant protein A, a structure similar to double-stranded DNA is formed, so that a topological structure of the slide surface is greatly changed, the LC orientation can be disturbed by the change, as a result, an LC optical image changing from darkness to colorfulness under the polarizing microscope is shown, and the pulmonary surfactant protein A can be detected; the LC biosensor is simple in structure and convenient and has quite high sensitivity and quite good selectivity, and with application of the method, the limit of detection of the pulmonary surfactant protein A is 5 nmol.L<-1>.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a nucleic acid aptamer liquid crystal biosensor for detecting pulmonary surfactant protein A, and a preparation method and a detection method of the liquid crystal biosensor. Background technique [0002] Pulmonary Surfactant Protein A (SP-A), a multifunctional glycoprotein, plays an important role in regulating pulmonary surfactant (PS) metabolism, gene expression, lung immunity and inflammatory response . SP-A is mainly formed in type II alveolar epithelial cells, with a small amount of expression outside the lung and high expression in the lung. The high or low expression level of pulmonary surfactant protein A is directly related to some clinical lung diseases, and at the same time it is also an indicator for the diagnosis and prognosis of some specific lung diseases. In the early stages of many lung diseases, it is difficult to assess the degree of damage to the alveolar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/23
CPCG01N21/23G01N33/6884
Inventor 王颖王兵李业芹邓世雄熊兴良
Owner JINING MEDICAL UNIV
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