Method for quantitatively detecting mercury ions in liquid sample and kit

A liquid sample, quantitative detection technology, applied in the field of molecular biology detection, achieves the effect of simple operation, avoiding adverse effects, and simplifying reaction steps

Active Publication Date: 2016-08-24
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, all of the above methods require serial dilution of samples with different gradients to obtain quantitative results. This process will be affected by a series of factors such as human operation.

Method used

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  • Method for quantitatively detecting mercury ions in liquid sample and kit
  • Method for quantitatively detecting mercury ions in liquid sample and kit
  • Method for quantitatively detecting mercury ions in liquid sample and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1 is used for the design of the biosensor sequence and primer and probe combination of mercury ion based on biosensor and microdroplet PCR quantitative detection

[0057] According to the reaction principle, the hairpin structure inside the biosensor is set, and different T-Hg-T structures are optimized, and different mercury ion biosensor sequences are designed: SEQ.1-3; the amplification primer probe set is designed: SEQ.4- 12.

Embodiment 2

[0058] Example 2 Establishment of a quantitative detection method based on biosensors and microdroplet PCR for mercury ion detection

[0059] 1.1 Experimental materials

[0060] Mercury ion standard samples and solutions used in this example were purchased from sigma company. Microdroplet PCR-related SuperMix and microdroplet generation chip were purchased from Bio-Rad Company, and enzymes and buffers for isothermal amplification were purchased from NEB Company.

[0061] 1.2 Biosensor isothermal extension

[0062] The isothermal amplification system is 2 μL of serum to be tested, 2 μL of 10 μM mercury ion biosensor, 2 μL of 10× isothermal PCR buffer, 2 μL of dNTP mixture, 0.2 μL of Klenow large fragment isothermal amplification enzyme, ddH 2 O to make up to 20 μL. When operating, first mix the three components of the solution to be tested, the biosensor, and water, and then incubate at 95°C for 5 minutes, then slowly cool down at room temperature, and after cooling down to ...

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Abstract

The invention provides a method for quantitatively detecting mercury ions in a liquid sample and a kit for quantitatively detecting the mercury ions. The absolute quantitative detection of the mercury ions can be performed through a quantitative detection method based on a biosensor and a droplet polymerase chain reaction (PCR) technology and the detection kit, the quantitative detection limit can reach 40 fmol, the sensitivity can reach 10 fmol, and the requirements of actually detecting of the mercury ions can be met. In addition, the method is simple to operate and high in flexibility and is an effective method for performing absolute quantitative detection of the mercury ions.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to a method and a kit for quantitatively detecting mercury ions in liquid samples. Background technique [0002] Among all heavy metal ions in water, mercury ion is one of all metal ions that are seriously harmful to human body. At this stage, a large number of detection methods for trace mercury ions have been developed. Recently, biosensor-based detection methods have achieved rapid progress due to their relatively simple operation and low cost. Most of the sensors are based on converting physical or chemical signals into nucleic acid signals, therefore, biosensors provide a method to detect heavy metal content by detecting nucleic acid signals. [0003] Based on the form of nucleic acid signal generation, biosensors can be classified into "turn-on" sensors and "turn-off" sensors. For "off" sensors, since the nucleic acid signal decreases with the increase o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2563/107C12Q2545/114
Inventor 罗云波许文涛黄昆仑朱鹏宇程楠田文营徐瑗聪
Owner CHINA AGRI UNIV
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