Fluorescent quantitative PCR method for rapid detection of imidacloprid water pollution

Abstract

The invention relates to a water body pollutant detection method, in particular to a rapid detection method for imidacloprid water body pollution by fluorescent quantitative PCR, belonging to the field of molecular biology. The present invention detects the mitochondrial gene of the aquatic insect Yixing Kuanjima wxya The technology of imidacloprid pollution monitoring based on the response of expression level to imidacloprid, the specific implementation steps are as follows: (1) cultivation of Yixing broad-based mayfia in water samples to be tested; (2) extraction of total RNA; (3) gel electrophoresis detection ; (4) Measurement of RNA concentration; (5) Inversion of RNA into cDNA; (6) Construction of standard products and creation of standard curve; (7) Measurement of gene expression; (8) Data analysis. The method has high sensitivity, good timeliness, strong specificity, simple operation, and can quickly detect whether the water body is polluted by imidacloprid.

Description

technical field [0001] The invention relates to a water body pollutant detection method, in particular to a rapid detection method for imidacloprid water body pollution by fluorescent quantitative PCR, belonging to the field of molecular biology. Background technique [0002] Imidacloprid belongs to neonicotinoid pesticide, which is a kind of neurotic insecticidal pesticide that can keep insects excited or paralyzed and then die. Its high toxicity to invertebrates, ease of use, flexibility, and long-term persistence make it a class of insecticidal pesticides with a high global market share. This type of pesticide represented by imidacloprid has lethal or sub-lethal effects on terrestrial and macrobenthic animals such as birds and bees, and aquatic invertebrates such as mayfly larvae, causing a decrease in earthworm fecundity and affecting ecological balance. At the same time, due to their good water solubility and high leaching rate, they are easy to enter the aquatic ecosy...

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem of imidacloprid water pollution detection, the present invention provides a rapid detection method for imidacloprid water pollution by fluorescence quantitative PCR, which measures the mitochondrial gene expression level of the aquatic insect Yixing broad-based mayfia by real-time fluorescence quantitative PCR technology to quickly detect whether the water body is affected by imidacloprid. Pollution, with the characteristics of convenience, high sensitivity, simple operation, etc.

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