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Method for rapidly, qualitatively and quantitatively detecting microcystic aeruginosa on spot and the reagent kit

A technology for quantitative detection of Microcystis aeruginosa, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and microorganism determination/inspection, can solve the problems of purifying algal cells and long detection time, and achieves simple and accurate operation. high sex effect

Inactive Publication Date: 2011-07-27
SUZHOU UNIV
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Problems solved by technology

[0005] The invention provides a kit and method for rapid on-site qualitative and quantitative detection of Microcystis aeruginosa, aiming to solve the problems of purifying algae cells and long detection time in the detection of Microcystis aeruginosa by existing molecular biology techniques

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  • Method for rapidly, qualitatively and quantitatively detecting microcystic aeruginosa on spot and the reagent kit
  • Method for rapidly, qualitatively and quantitatively detecting microcystic aeruginosa on spot and the reagent kit
  • Method for rapidly, qualitatively and quantitatively detecting microcystic aeruginosa on spot and the reagent kit

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Embodiment 1

[0046] Example 1: A kit and method for on-site rapid qualitative and quantitative detection of Microcystis aeruginosa

[0047] The present invention uses the transcription unit spacer ITS (Internal Transcribed Spacer) between the ribosome subunits of Microcystis aeruginosa as the target sequence, and searches for a highly specific DNA sequence in the target fragment as a probe. The key to the qualitative analysis of microalgae in natural water blooms using whole-cell fluorescence in situ hybridization is to find suitable DNA fragments in the genome of Microcystis aeruginosa as probes. This DNA segment must be highly variable, that is, the sequence behaves differently between different species and even between different geographical strains of the same species. ITS sequences can be considered as FISH probes due to their high variability.

[0048] Microcystis aeruginosa (photographs under natural light conditions are attachedfigure 1 (shown) is the main species of bloom microal...

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Abstract

The invention discloses a reagent kit and a method for rapidly, qualitatively and quantitatively detecting microcystic aeruginosa on spot, detecting by using a nucleic acid probe with a fluorescein isothiocyanate added at the 5'end. The detection method comprises the following steps: firstly fixing algae cells; dehydrating and hybridizing, and cleaning and observing the algae cells under a fluorescence microscope.The invention has the advantages of correctness, rapidness and simpleness, and has simple operation because less than 20 hours is required for the whole detection process. The most important is that detection can be performed on spot, and qualitative analysis of the algae cells can be carried out in natural water body, which can not be achieved by the prior other methods for authenticating microcystis flos-aquae, therefore, the invention has favorable application prospect on detection of the microcystis flos-aquae.

Description

technical field [0001] The present invention relates to a kit and method for on-site rapid qualitative and quantitative detection of water bloom microalgae, in particular to a whole-cell fluorescence in situ hybridization technique (Whole-cell Fluorescence in situ Hybridization, FISH) for early warning and forecasting of water blooms Method for detection of Microcystis aeruginosa. Background technique [0002] Fluorescence in situ hybridization technology is the product of the combination of cell biology and molecular biology technology (mainly molecular hybridization technology). The probe hybridizes (combines) with the DNA of the chromosome or nucleus of the metaphase cell, and reveals the position of the DNA fragment homologous to the fluorescent probe on the chromosome, thereby locating the DNA fragment of this known sequence in the cell. Fluorescence in situ hybridization technology is not only accurate, fast, sensitive, but also simple and convenient. It has been wide...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N21/64C12R1/89
Inventor 沈颂东王文杰赵辰红苏量张冬梅
Owner SUZHOU UNIV
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