Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe

A nucleic acid aptamer and test strip technology, which is applied in measuring devices, recombinant DNA technology, DNA/RNA fragments, etc., can solve problems such as inability to use on-site detection, limit widespread use, and high detection cost, and achieve application Broad range, high specificity, low cost effect

Inactive Publication Date: 2014-08-13
HEFEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although relying on traditional instruments for detection has the advantages of high detection sensitivity and good test result stability, large-scale instruments are usually expensive and require professional operators with relevant knowledge. At the same time, there are disadvantages such as high detection costs, which limit the large-scale instruments Scale detection and on-site rapid detection
The enzyme-linked immunosorben

Method used

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  • Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe
  • Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe
  • Probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of probe

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preparation example Construction

[0030] One aspect of the present invention also provides a method for preparing a bisphenol A immunochromatographic gold standard test strip, comprising:

[0031] Colloidal gold is reacted with the nucleic acid aptamer probe in the solution to form a nucleic acid aptamer probe-gold nanoparticle complex, and then the nucleic acid aptamer probe-gold nanoparticle complex is applied to the binding On the pad substrate, a nucleic acid aptamer probe-gold label binding pad is formed;

[0032] And, the bisphenol A-bovine serum albumin conjugate solution and the nucleotide chain solution capable of complementary pairing with the nucleic acid aptamer probe are respectively applied to the coated membrane substrate to form a detection line and a reference line independent of each other, Obtain the coating film;

[0033] And, at least the sample membrane, the nucleic acid aptamer probe-gold label binding pad, the coating film and the water-absorbing film are bonded to the liner in sequenc...

Embodiment 1

[0049]The specific operation steps of the preparation method of a bisphenol A immunochromatographic gold standard test strip based on nucleic acid aptamer probes involved in this embodiment are as follows:

[0050] (1) Select a 25-40nm colloidal gold solution, centrifuge at a centrifugal speed of 9500r / min for 7 minutes, discard the supernatant to obtain a 10-fold concentrated colloidal gold solution A;

[0051] Add 9 μL of 1 μM / L nucleic acid aptamer probe (abbreviated as ssDNA below) to 1 μL of acetate buffer solution and 1.5 μL of 1 μM / L TCEP for 1 h in the dark to obtain mixed solution B;

[0052] Add 100 μL of 10-fold concentrated colloidal gold solution A (concentration about 100 mM) to the above mixture B, and shake at room temperature for 30 minutes; add 85 μL of 100 μM / L dATP to the above solution, and shake at room temperature for 15 minutes; the shaking is completed Then add 20 μL of 0.1 M / L NaCl solution, let stand at room temperature for 30 min, and then put it in...

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Abstract

The invention discloses a probe capable of specifically identifying bisphenol A nucleic acid aptamer and test strip detection application of the probe. The probe has a sequence shown in SEQID NO.1, and can be applied to the preparation of a bisphenol A immunochromatographic gold-labeled test strip, and the like; the test strip comprises a lining plate; at least four sticking objects including a sample film, a nucleic acid aptamer probe-gold-labeled conjugate pad, an enveloping film provided with a detection line and a reference line, and a water absorption film are sequentially stuck on the lining plate, wherein the nucleic acid aptamer probe-gold-labeled conjugate pad comprises compound of a nucleic acid aptamer probe and gold nanoparticles. Due to the test strip, the rapid qualitative and quantitative determination of bisphenol A can be realized by direct observation without using other auxiliary instruments and equipment, the detection work can be rapidly completed, and the accurate detection result can be obtained; the probe capable of specifically identifying the bisphenol A nucleic acid aptamer has the characteristics of being high in specificity, good in stability, low in cost, easy to use and the like, and can be used for a hospital, a family, the customhouse with responsibility of food safety inspection and quarantine, an airport, a port and the like which need a rapid field detection mechanism.

Description

technical field [0001] In particular, the present invention relates to a specific recognition bisphenol A nucleic acid aptamer probe and its application in detecting bisphenol A, for example, a bisphenol A immunochromatographic gold standard test strip based on the nucleic acid aptamer probe , its preparation method and application. technical background [0002] With the development of science and technology and the increasing requirements for the convenience and speed of life, plastic products have been widely used in all aspects of life. Bisphenol A is an important component of synthetic polycarbonate and epoxy resins, which are used to make plastic (milk) bottles, suction cups for children, and coatings on the inside of food and beverage (milk powder) cans. According to research, bisphenol A can lead to endocrine disorders, induce precocious puberty, and threaten the health of fetuses and children. At the same time, cancer and obesity caused by metabolic disorders are al...

Claims

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Application Information

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IPC IPC(8): G01N33/558
CPCG01N33/54353C12N15/115G01N33/558
Inventor 钟友好陈伟赵弟萍薛峰姚丽
Owner HEFEI UNIV OF TECH
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