Kit capable of simply detecting various red-tide algae in natural water samples

Abstract

The invention discloses a kit capable of simply detecting various red-tide algae in natural water samples. The kit comprises membrane classification gene chip strip, species specificity probes containing detected Heterosigacarterae, Prorocentrum donghaiense, Nitzschia closterium, Symbiodinium, skeletonema costatum, Chaetoceros debilis, Prorocentrum triestinum Schiller and Alexandrium tamarense, a negative control probe, a positive control probe, a reagent for preparing nucleic acid, and a reagent for nucleic acid hybridization, wherein each probe is used for detection for twice. The detection kit provided by the invention is portable, sensitive and rapid, can simultaneously detect the red-tide algae, such as Heterosigacarterae, Prorocentrum donghaiense, Nitzschia closterium, Symbiodinium, skeletonema costatum, Chaetoceros debilis, Prorocentrum triestinum Schiller and Alexandrium tamarense, in the natural samples without expensive instruments, and can realize on-site detection on the various red-tide algae in the natural samples.

Description

technical field [0001] The invention relates to the technical field of marine biology, in particular to a kit for simply detecting various red tide algae in natural water samples. Background technique [0002] Red tides are the discoloration of sea water due to the overgrowth or accumulation of marine plankton, including algae (the main species), protozoa and bacteria. In recent years, due to the intensification of human activities, the ocean has become increasingly eutrophic, resulting in frequent red tides, which have become a global ecological disaster, causing huge losses to the aquaculture industry, fishing industry and coastal tourism. The marine ecological environment has a negative impact and even directly threatens human health. [0003] Harmful red tides in my country show a trend of increasing outbreak frequency, expanding area, prolonging duration and intensifying harm. For example, in 2004, the number of red tides broke out in my country reached 120 times, esp...

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Problems solved by technology

[0007] (1) Red tide algae are generally small and difficult to identify. Light microscopy can only identify the genus sometimes. To further determine its specific species, electron microscopy observation may be required. Therefore, the work is cumbersome and the detection cost is high, and it is difficult to adapt to daily water. Environmental monitoring requires fast and accurate processing of a large number of samples;

[0008] (2) The occurrence of red tide is usually an explosive event. The concentration of red tide algae in natural water bodies is low in the early stage of red tide, and it is mixed with non-red tide algae. Some conventional monitoring methods, such as algae collection, qualitative and Quantitative records are more difficult, which may delay the early warning of red tide algae;

[0009] (3) The appearance of cells of red tide algae can easily change with the living environment and physiological state, so its identification is highly dependent on experience, which makes the identification of some red tide microalgae the expertise of taxonomists, and also increases the natural Difficulty in identification, monitoring and early warning of red tide provenance in sea areas

[0012] (1) Existing molecular detection technologies, except gene chip technology, can only identify and detect a single species of red tide algae in natural samples, but cannot detect multiple species of red tide algae in natural samples;

[0013] (2) On-site detection cannot be realized: Existing molecular detection methods must rely on special experimental instruments, such as DNA sequencing requires a PCR instrument and a sequencer, restriction fragment length polymorphism and random amplified polymorphic DNA require a DNA electrophoresis instrument , Whole-cell fluorescence in situ hybridization requires a fluorescence microscope, real-time fluorescent quantitative PCR requires a fluorescent quantitative PCR instrument, and sandwich hybridization requires a microplate reader, etc., which makes these detection methods must be completed in the laboratory, and cannot be used for red tide algae in natural water bodies. Conduct on-site testing

[0014] (3) Disadvantages of the existing chip detection technology: Although the existing chip technology can realize the parallel detection of various red tide algae in natural samples, its nucleic acid preparation requires a nucleic acid amplification instrument, and the interpretation of the results requires a fluorescent scanner. Its detection cost is high, and it is difficult to become an on-site detection technology

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