Method for detecting blue algae capable of producing 2-MIB by PCR qualification and TaqMan fluorescent quantitative PCR quantity

Abstract

The invention discloses a method for detecting blue algae capable of producing 2-MIB by PCR qualification and TaqMan fluorescent quantitative PCR quantity. The method comprises the following steps: water sample to be measured is collected, filtered through a cellulose acetate film, extracted and purified total genome DNA on the filtered algae; B, a qualitative PCR reaction system comprises: corresponded monoterpene cyclase gene is synthesized, 2-MIB blue algae is taken as positive control and double distilled water is taken as negative control; C, a qualitative PCR reaction progress comprises: denaturation is carried out at the temperature of 94 DEG C for 3 minutes, circulated and finally extended; D, qualitative PCR products and positive control, negative control; E, the fluorescent quantitative PCR reaction system, aseptic double distilled water is added in negative template control; F, the fluorescent quantitative PCR reaction progress; G, stand curve making; H, the concentration of blue algae capable of producing 2-MIB in the water sample is calculated according to the Ct value and the standard curve of the sample to be measured. The method of the invention has the advantages of easy process, simple operation, high sensitivity and accuracy, so that the concentration of blue algae capable of producing 2-MIB in the water sample can be rapidly and accurately quantified.

Description

technical field [0001] The present invention relates to the fields of water environment monitoring, water ecology, microbial harmful metabolites, etc., and more specifically relates to a method for detecting 2-MIB-producing cyanobacteria in the environment by using PCR (polymerase chain reaction) and fluorescent quantitative PCR. It is suitable for early warning, rapid qualitative and quantitative analysis of algae-derived 2-MIB in drinking water sources such as reservoirs, lakes and rivers, and aquatic ecosystems. Background technique [0002] In recent years, with the increasing eutrophication of water bodies, cyanobacterial blooms have broken out in large areas all over the world, especially in my country. The water bloom problem has become a major problem in the field of environmental protection in our country. The hazards of cyanobacterial blooms are manifested in many aspects, among which algal toxins, which are a direct threat to human health and drinking water, have...

AI Technical Summary

Problems solved by technology

Ordinary PCR detection technology can qualitatively detect whether cyanobacteria produce 2-MIB and whether there are 2-MIB cyanobacteria in the environment, but cannot quantitatively detect the concentration of 2-MIB cyanobacteria in water

Images