Electrochemistry ampere detection method for quantitively detecting polymerase chain reaction (PCR) based on methylene blue indicator

Abstract

The invention discloses an electrochemistry ampere current detection method for quantitively detecting a polymerase chain reaction (PCR) amplification product based on a methylene blue deoxyribonucleic acid (DNA) indicator. The method is applied to the PCR product quantitive detection of a mitochondria actual sample of a normal person. The method includes that the design and synthesis of primers required by the PCR amplification are performed according to amplified fragments of genes to be detected; a mitochondria gene sequence of the normal person is used as an example; a preprocessed glassy carbon electrode is placed in a PCR system containing amplification specimens, and a certain quantity of DNA hybridization indicator methylene blue is added in the system; and corresponding PCR reaction conditions are set according to target segments, the amplification is performed, and simultaneously current signal changes of the methylene blue are collected according to an electrochemistry current-time curve method so that the electrochemistry detection of the PCR amplification product is achieved. The method is high in selectivity and sensitivity, the quantitive detection of the PCR product is achieved, and compared with traditional methods, and the method is fast, simple, convenient and economical.

Description

technical field [0001] The invention relates to an electrochemical amperometry method for quantitative detection of PCR amplification products based on methylene blue DNA indicator technology. Background technique [0002] With the development of molecular biology and biotechnology, especially the implementation of the Human Genome Project, people have a deeper understanding that as the carrier of genetic information, the structural changes of DNA will have a significant impact on organisms, and are related to various Causes of genetic diseases. Therefore, people's DNA testing methods are becoming more and more important. Among gene analysis technologies, polymerase chain reaction (PCR) technology has become an important means of gene diagnosis and analysis due to its advantages of simplicity, speed and sensitivity. However, there are two major problems that need to be solved urgently in PCR technology: one is that it cannot be quantified, and the other is false positives ...

AI Technical Summary

Problems solved by technology

However, there are two major problems that need to be solved urgently in PCR technology: one is that it cannot be quantified, and the other is false positives caused by pollution.

Studies have shown that under the optimal conditions, the efficiency of symmetric PCR amplification in liquid phase can reach about 90%, while the efficiency of asymmetric PCR amplification is only 70%, and the efficiency of symmetric PCR amplification on solid phase is also higher than that in liquid phase. In-phase symmetric PCR amplification efficiency is 20-30% lower

Images