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A detection method and high-sensitivity technology, applied in the biological field, can solve the problems of complicated experimental operation, high experimental cost, damage to deoxyribozyme activity, etc., and achieve the effects of obvious color development, low cost and mature reaction conditions.
Active Publication Date: 2020-10-27
张睿
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Although these methods in Pb 2+ Great contributions have been made in detection, but most of these methods are based on the labeling and modification of deoxyribozymes, or use more expensive materials such as gold nanometers. The activity of the enzyme causes some damage, so a rapid and sensitive detection method that does not require group labels and gold nanomaterials is needed
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[0025] The present invention is illustrated below with specific examples.
[0026] A high-sensitivity detection method for lead ions includes the establishment of a linear regression equation and the detection of samples to be tested.
[0027] The establishment of described linear regression equation may further comprise the steps:
[0028] (1) Preparation of standard solution: Dilute the lead acetate solution to different concentrations of 0-60 μM to obtain the standard solution;
[0029] (2) Primer preparation: add the standard solution to GR-5 deoxyribozyme to a volume of 40 μL, incubate at 25°C for 3 minutes, then add 5 μL of stop solution, mix well, and store at 4°C to obtain the single-stranded primer X The preparation method of the GR-5 deoxyribozyme comprises the following steps: taking respectively 4 μL of GR-5-substrate chain whose molar concentration is 10 μM, and 4 μL of GR-5-enzyme chain whose molar concentration is 10 μM, adding buffer and Water to 40μL, so tha...
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Abstract
The invention mainly relates to the field of biotechnology and discloses a high-sensitivity lead ion detection method. The high-sensitivity lead ion detection method comprises the following steps: sample processing, preparation of a primer, complementary hybridizing, isothermal amplification, preparation of a mimic enzyme, catalytic color development, and calculation of the concentration of lead ions. The operation is simple, the principle is clear, reaction conditions used at each step are mature and easy to control, meanwhile, fluorescence and gold nanoparticle groups do not need to be labelled, the reduction of the activity of dnazyme is avoided, expensive agents and large instruments are not used, the cost is low, the reaction time is relatively short, the total reaction time is less than 2 hours, meanwhile, the detection of the lead ion is relatively sensitive, the minimum detection concentration is 10 nM, a water body can be strictly monitored, the environmental pollution is under control, the safety of the human body is guaranteed, meanwhile, the color development is obvious and is visible to naked eyes, and the novel lead ion detection method is provided.
Description
technical field [0001] The invention mainly relates to the field of biotechnology, in particular to a highly sensitive lead ion detection method. Background technique [0002] Lead ions can cause harm to the human body at very low concentrations, and can produce side effects at very low concentrations. After lead enters the human body, it will cause multiple damages to the nervous system, blood circulation system, digestive system, and urinary system. . The World Health Organization stipulates that the lead content in drinking water should not exceed 10 μg / L, and the lead content in the air should not exceed 0.5 μg / m 3 , the general human body contains about 77mg of lead. The Food and Agriculture Organization of the United Nations and the World Health Organization proposed that the permissible daily lead intake per person is about 420μg, and the lead poisoning standard is less than 100μg / L. When the blood lead content of the human body is greater than or equal to 100μg / L L...
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