Cleavage type universal-partition ultrafast-amplification visual sensor for zinc ions
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A technology of zinc ions and sensors, applied in the field of biosensors, can solve the problems of long detection cycle, high price, complicated pre-treatment, etc.
Active Publication Date: 2018-12-07
CHINA AGRI UNIV
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[0003] Traditional zinc ion detection methods can generally be divided into cold atomic absorption spectrometry, graphite carbon atomic absorption spectrometry and flame atom
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Embodiment 1
[0095] Example 1 The establishment of a zinc ion-cutting universal partition ultrafast amplification visualization sensor
[0096] 1. Experimental materials
[0097] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, zinc chloride, 2,2-azino-bis(3-ethyl- Benzothiazole-6-sulfonic acid) diamine salt (ABTS), H2O2, 4-hydroxyethylpiperazineethanesulfonic acid (HEPES), sodium hydroxide, disodium hydrogen phosphate, all purchased from Thermo Fisher Scientific (Thermo Scientific Life Technologies). All experimental water was from Milli-Q pure water system.
[0098] The sequence design is as follows (SEQ ID NO:1-7):
[0099]
[0100] Note: The ribozyme cuts the target product and the 3' end sequence of the amplified template is complementary;
[0101] The cleavage site is indicated by "-";
[0102] The ribozyme substrate chain-Zn and the ribozyme enzyme chain-Zn together constitute a z...
Embodiment 2
[0122] The specificity investigation of embodiment 2 sensor
[0123] According to the biosensor constructed in Example 1, 500nM Zn 2+ , 10μM Ag + , Mg 2+ , Ca 2+ 、Cd 2+ , Hg 2+ 、Cr 3+ Added to the system for detection, the results show that the established Zn 2+ The biosensor has good specificity ( Image 6 ).
Embodiment 3
[0124] Embodiment 3 standard addition experiment
[0125] Get high-purity water and detect with the biosensor constructed in embodiment 1, Zn 2+ No detection was carried out, and the standard addition experiment was carried out on it, and the results obtained by continuous measurement are shown in Table 1.
[0126] Table 1 Zn 2+ Spike recovery test results
[0127]
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Abstract
The invention provides a cleavage type universal-partition ultrafast-amplification visual sensor for zinc ions. The visual sensor comprises (1) a cleavage system, (2) a sPCR amplification system, (3)a HCR system, and (4) a detection system containing an ABTS developing solution. According to the invention, primers, a template and probes are ingeniously designed, so ultrafast amplification of thetemplate can be realized in the presence of zinc ions, and an amplification product is allowed to form a G-quadruplex in a proper environment by triggering a HCR reaction; and color development is performed by utilizing the peroxidase-like activity of the G-quadruplex. Thus, the problem of difficulty in realizing visual detection of traditional PCR products is overcome, and rapid and visual detection of zinc ions is realized. Moreover, the sensor provided in the invention has the characteristics of high specificity and high sensitivity to zinc ions, and is more objective and accurate in detection results.
Description
technical field [0001] The invention relates to the technical field of biosensors, in particular to a zinc ion-cutting universal partition ultrafast amplification visualization sensor. Background technique [0002] Zinc, the chemical symbol is Zn, the atomic number is 30, and the atomic weight is 65.38. It belongs to group IIB and is a light gray transition metal. It is widely distributed in nature and mainly exists in the state of zinc sulfide and zinc oxide. It can also be combined with Mineral symbiosis of many elements such as lead, copper, and zinc. Zinc is the most abundant trace element in the human body. Its content is as high as 3g. It mainly participates in the metabolism of the body in the form of zinc ions, and participates in the synthesis and activation of more than 200 enzymes in the human body. It is an essential substance in the body's metabolism. Zinc pollution refers to the environmental pollution caused by zinc and its compounds. The main sources of pol...
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