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A probe combination, chip, kit and method for detecting toxin-producing Microcystis bacteria type

A Microcystis type, Microcystis technology, applied in the field of microbial detection, to achieve simple and convenient data processing, simple operation methods, and the effect of making up for insufficient resolution

Active Publication Date: 2022-07-26
壹健生物科技苏州有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Polycystis

Method used

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  • A probe combination, chip, kit and method for detecting toxin-producing Microcystis bacteria type
  • A probe combination, chip, kit and method for detecting toxin-producing Microcystis bacteria type
  • A probe combination, chip, kit and method for detecting toxin-producing Microcystis bacteria type

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Screening of Toxigenic Microcystis Type-specific Detection Probes

[0058] In this example, the genome sequences of 11 strains were used for probe design. 11 strains including Microcystiswesenbergii FACHB-929, Microcystis aeruginosa FACHB-909, Microcystis aeruginosaFACHB-911, Microcystis aeruginosa FACHB-925, Microcystis aeruginosa FACHB-975, Microcystis aeruginosa FACHB-978, Microcystis sp.FACHB-10105, Microcystissp , Microcystis sp.FACHB-1026, Microcystis sp.FACHB-1027 and Microcystis elabens FACHB-917. The 11 strains were all derived from the Freshwater Algae Culture Collection at the Institute of Hydrobiology (FACHB), and all obtained genomes sequence.

[0059] The following is the process of designing a specific detection probe for the toxigenic strain of Microcystis wesenbergii FACHB-929 using the genome sequence of the toxigenic Microcystis strain Microcystis wesenbergii FACHB-929 (GCA_014698625.1).

[0060] 1. Dataset collection:

[0061] To design ...

Embodiment 2

[0075] Example 2 Preparation of gene chip for specific detection of Toxigenic Microcystis

[0076] Using the methods of the examples, Microcystis aeruginosa FACHB-909, Microcystis aeruginosa FACHB-911, Microcystis aeruginosa FACHB-925, Microcystis aeruginosa FACHB-975, Microcystis aeruginosa FACHB-978, Microcystis sp.FACHB-1005, Microcystis sp.FACHB-1023, Probes were designed for the genome sequences of Microcystis sp. FACHB-1026, Microcystis sp. FACHB-1027 and Microcystis elabens FACHB-917, and the results are shown in Table 2.

[0077] Table 2 Specific probes for 10 toxin-producing bacteria types

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088] Thus, the number of designed probes is shown in Table 3:

[0089] Table 3 Probes corresponding to each toxin-producing bacteria type

[0090] Toxigenic bacteria Number of probes SEQ ID No. Microcystis wesenbergii FACHB-929 47 1～47 ...

Embodiment 3

[0120] Embodiment 3 Utilize the gene chip prepared in embodiment 2 to quickly detect the method for the toxin-producing bacteria type of common water body toxin-producing Microcystis

[0121] 1. Target sample DNA extraction

[0122] Collect water from lakes or rivers and filter the cyanobacteria to enrich the cyanobacteria, and use the enriched material as the sample to be tested. Then, use the DNA extraction kit of Agilent CGH MicroArray to extract nucleic acid from the sample to be tested. The nucleic acid extraction of the sample to be tested is carried out by conventional methods in the field.

[0123] 2. DNA purification

[0124] The main steps of DNA purification are:

[0125] (1) Take 500ng of the extracted DNA sample in the interrupted tube, make up H 2 0 to 50 μL. Set the interruption time to 90s;

[0126] (2) Pre-equilibrate the OnePure MagBeads at room temperature for 30 minutes, and fully shake and mix to ensure that there is no obvious magnetic bead precipita...

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Abstract

The invention discloses a probe combination for detecting toxin-producing Microcystis bacteria type, and belongs to the technical field of microorganism detection. Wherein, the probe combination includes probes for detecting each toxin-producing Microcystis species in the toxigenic Microcystis species combination. The invention also discloses a gene chip, a kit and a detection method prepared by using the above probe combination. The present invention has the dual advantages of high flux and high resolution, and the detection method of the present invention is simple in operation, and data processing is simpler and more convenient, so that the present invention has important application value for long-term dynamic monitoring of cyanobacterial pollution in water bodies. It can guide algal bloom control and water ecological restoration.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular, relates to a probe combination, a chip, a kit and a method for detecting toxin-producing Microcystis bacteria. Background technique [0002] With the continuous development of human industrial activities and social economy, algal pollution caused by eutrophication of lakes has become a major global environmental problem. Eutrophication of water mainly refers to the phenomenon of rapid reproduction of plankton in water and the decrease of dissolved oxygen in water caused by the accumulation of nitrogen and phosphorus in water. Eutrophication of water bodies will bring about three main hazards. The first is to affect the water ecosystem. When the nutrients in the water are too rich, algae over-proliferate and the water surface is covered with algae, which affects the photosynthesis of organisms in the lower water. At the same time, the decomposition of dead algae...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6837C12Q1/6895C12Q1/04C12N15/11
CPCC12Q1/6837C12Q1/6895C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/113C12Q2565/501
Inventor 束浩月林德春金桃蒋华
Owner 壹健生物科技苏州有限公司