A Chromium-Ion Cutting General-Purpose Partition Ultrafast Amplification Visualization Sensor
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A technology of chromium ions and sensors, applied in the field of biosensors
Active Publication Date: 2020-07-28
CHINA AGRI UNIV
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However, these methods rely on large-scale instruments and dedicated personnel to operate, and are limited in on-site and field detection, requiring complex pretreatment of samples. Therefore, it is urgent to develop non-polluting, simple, fast, high-sensitivity and high-specificity methods to Meet the needs of trace metal detection
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Embodiment 1
[0081] Example 1 Establishment of a chromium ion cutting type universal partition ultra-fast amplification visual sensor
[0082] 1. Experimental materials
[0083] SYBR Gold nucleic acid dye, nucleic acid molecular weight standard ultra-low range DNA ladder, dNTP, ExTaq DNA polymerase, 10×Taq buffer, hemin, chromium chloride, 2,2-diazo-bis(3-ethyl- Benzothiazole-6-sulfonic acid) diamine salt (ABTS), H2O2, morpholine ethanesulfonic acid monohydrate (MES), potassium chloride, sodium chloride, magnesium chloride, potassium hydrogen phosphate, disodium ethylenediaminetetraacetic acid , Urea, were purchased from Thermo Scientific Life Technologies (Thermo Scientific Life Technologies). All experimental water comes from Milli-Q pure water system.
[0084] The sequence design is as follows (SEQ ID NO:1-5):
[0085]
[0086] Note: Ribozyme cleaves the target product and the 3'end sequence of the amplification template is complementary;
[0087] The bold base sequence added at the end of the...
Embodiment 2
[0106] Example 2 Investigation of sensor specificity
[0107] According to the biosensor constructed in Example 1, 1nM Cr 3+ , 10μM Cu 2+ , Zn 2+ , Cd 2+ , Hg 2+ , Na + , Ag + , Added to the system for testing, the results show that the established Cr 3+ Biosensors have good specificity ( Figure 4 ).
Embodiment 3
[0108] Example 3 Standard addition experiment
[0109] Take high-purity water and use the biosensor constructed in Example 1 to detect, Cr 3+ If no detection was found, it was subjected to a standard addition experiment, and the results obtained from consecutive measurements are shown in Table 1.
[0110] Table 1 Cr 3+ Result of spike recovery experiment
[0111]
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Abstract
The invention provides a cleavage type universal-partition ultrafast-amplification visual sensor for chromium ions. The visual sensor comprises (1) a cleavage system, (2) a sPCR amplification system and (3) a detection system containing an ABTS developing solution. According to the invention, primers, a template and probes are ingeniously designed, so ultrafast amplification of the template can berealized in the presence of chromium ions, and an amplification product is allowed to form a G-quadruplex in a proper environment; and color development is performed by utilizing the peroxidase-likeactivity 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 chromium ions is realized. Moreover, the sensor provided in the invention has the characteristics of high specificity and high sensitivity to chromium 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 chromium ion cutting type universal cut-off ultra-fast amplification visualization sensor. Background technique [0002] Chromium symbol Cr, silver-white metal, belongs to group VIB in the periodic table of elements, chromium has atomic number 24, atomic weight 51.996, body-centered cubic crystal, and common valences are +3, +6 and +2. The average content of chromium in the earth's crust is 0.010% to 0.011%, and the distribution is extremely uneven. my country's chromium resources are not abundant, and trivalent chromium is one of the indispensable trace elements in the human body. Adults need about 500-700 micrograms of chromium per day. When chromium is deficient, the human body will easily suffer from diabetes and coronary atherosclerosis, which will affect the fat metabolism in the body. However, too much chromium can also cause pollution. The toxicity of hexavalent chromi...
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