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Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit

A technology for detection kits and detection methods, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of difficulty in obtaining sensitivity and detection range, cumbersome synthesis and preparation of nano-gold, and insufficient sensitivity. High sensitivity and detection range, low requirement, high sensitivity effect

Inactive Publication Date: 2013-09-18
XI AN JIAOTONG UNIV +1
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Problems solved by technology

However, although the results of colorimetry are visible to the naked eye, the sensitivity is not high enough, and the synthesis and preparation of nano-gold are also relatively cumbersome.
However, the electrochemical method has high sensitivity, but the operation is complicated and takes a long time, and it needs to be equipped with expensive electrochemical related instruments. Satisfactory sensitivity and detection range

Method used

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  • Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit
  • Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit
  • Lead ion detection kit based on constant-temperature cascade nucleic acid amplification and detection method of lead ion detection kit

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Embodiment 1

[0058] Embodiment 1 is based on deoxyribozyme, chain displacement amplification and nickase amplified fluorescent signal detection method for lead ion concentration in river water, the concentration of lead ion in the complex system is detected, the specific operations are:

[0059] 1) Handle the solution to be tested

[0060] Let the collected river water solution stand still, filter out suspended particles with a micro-membrane with a pore size of 0.2 μm, and add 1×10 -11 mol, 10×10 -11 mol,100×10 -11 mol standard lead ion solution for use.

[0061] 2) Using lead ions in the solution to trigger constant temperature cascade nucleic acid amplification to amplify the fluorescent signal

[0062] Add 2 μL of the above lead ion solution to the solution containing 20 nM deoxynucleic acid, 10 nM SDA template, 50 nM hairpin molecule H, 250 nM molecular beacon, 2.5 U / μL Klenow fragment polymerase, 4 U Nt.BbvCI nuclease nickase, and 0.2 mM dNTPs 1×NEB buffer2 (10mM Tris-HCl, 50mM N...

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Abstract

The invention discloses a lead ion detection kit based on constant-temperature cascade nucleic acid amplification and a detection method of the lead ion detection kit. Lead ions are recognized through deoxyribozyme, strand displacement amplification and a single chain triggered DNA (deoxyribonucleic acid) molecular machine are combined to realize constant-temperature cascade nucleic acid amplification, and the concentration of the lead ions is converted into a remarkably-amplified fluorescence detection signal, thus the concentration of trace lead ions can be precisely measured. Compared with other methods based on deoxyribozyme, the detection method is rapider and higher in sensitivity; and in addition, the requirement for equipment is lower, and the operation is simpler.

Description

technical field [0001] The invention belongs to the technical field of lead ion detection, and relates to a lead ion detection kit based on constant temperature cascade nucleic acid amplification and a detection method thereof. Background technique [0002] Lead ion is a common and highly toxic pollutant that can adversely affect plants and animals in aquatic ecosystems. The main sources of lead contamination are probably gasoline and lead-based paints. For humans, lead ions can cause many health problems such as anemia, kidney failure and muscle paralysis. Even at low concentrations, lead poisoning can cause serious damage to the brain and central nervous system in children. Therefore, it is very important to develop an ultrasensitive and highly selective lead ion detection method for clinical detection, industrial and environmental monitoring. [0003] The traditional main methods for detecting heavy metals such as lead ions are chromatography, atomic absorption / emissio...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/34
Inventor 赵永席张青袁慧王芳霞董绘阳白凯
Owner XI AN JIAOTONG UNIV
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