Real time fluorescent quantitative PCR detection method for blue algae producing microcystic toxins

Abstract

The invention relates to a realtime fluorescent quantitative PCR detection method of cyanobacteria producing microcystin (MC) for rapidly, accurately and quantitatively detecting cyanobacteria producing microcystin (MC) in eutrophic water body of a freshwater lake. The method comprises the following steps of: collecting and processing a lake water sample and extracting the total DNA; carrying out realtime fluorescent quantitative PCR detection of mcyA-CD gene fragment by using SYBR Green I chimeric fluorescence method; recording the fluorescence values and C(t) values of a standard curve sample, a positive reference, a negative reference and a sample to be detected; drawing a standard curve by a computer; and calculating the copy number of the mcyA-Cd gene fragments in the sample to be detected and the concentration of cyanobacteria producing MC. The inventive method can rapidly and accurately detect concentration of cyanobacteria producing MC in eutrophic water body of freshwater lake and has good repeatability.

Description

technical field [0001] The invention relates to a real-time fluorescent quantitative PCR detection method for microcystin-producing cyanobacteria, which uses real-time fluorescent quantitative PCR (polymerase chain reaction) technology to quantitatively detect microcystin-producing cyanobacteria in water bodies, and belongs to the technical field of molecular detection. Background technique [0002] The eutrophication of lake water will lead to the outbreak of cyanobacteria bloom, which has become one of the major environmental pollution problems faced by our country and the world. Some cyanobacteria in cyanobacterial blooms can produce algal toxins, among which microcystin (MC) is the most important and common hepatotoxin in fresh water, which has a serious impact on aquatic organisms, the safety of human drinking water and human health. Studies have shown that people living in areas with high incidence of cyanobacteria blooms have a significantly higher incidence of liver ...

AI Technical Summary

Problems solved by technology

[0005] Amplification of the mcyA-Cd fragment by general PCR technology can only qualitatively distinguish whether cyanobacteria produce MC, but cannot quantitatively detect the concentration of MC-producing cyanobacteria in water

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