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Detection probe for minisize Prorocentrum

A technology for detecting probes and Prorodinova, which is applied in the field of specific probes and oligonucleotide probes. It can solve the problems of difficult operation, large fluctuation of detection results, and small number of captured probes, so as to achieve easy execution. Effect

Inactive Publication Date: 2006-12-06
SHANGHAI JIAOTONG UNIV +1
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Problems solved by technology

Its main disadvantages are: first, because the length of the capture probe is only 10-30 bp, the number of capture probes that can effectively distinguish close relatives of microalgae is very small; second, it is necessary to prevent RNA degradation during the entire detection process, However, various steps such as washing and antibody reaction can easily cause RNA degradation, so the operation is difficult and the test results fluctuate greatly

Method used

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  • Detection probe for minisize Prorocentrum
  • Detection probe for minisize Prorocentrum

Examples

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Embodiment

[0030] Example: Qualitative and quantitative detection of Prorocentrum micronii using S1 enzyme protection analysis for rRNA and sandwich hybridization technology

[0031] In this example, the combination of S1 enzyme protection analysis technology and sandwich hybridization technology is used to realize the qualitative and quantitative detection of the miniature red tide organism Prorocentrum minimum (Prorocentrum minimum), and the steps are as follows:

[0032]1.1 The search for the specific region of Prorocentrum miniature rRNA and the design and synthesis of the S1 enzyme protection analysis probe

[0033] Through the determination of the ribosomal large subunit 28S rRNA sequence of Prorocentrum microphylla and multiple sequence comparisons with the rRNA sequences of other algae, it was found that the 430-520 region of the large subunit rRNA was different from other algae, and the sequence of this region was determined to be a characteristic sequence ; Design and synthesiz...

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Abstract

A detection probe for Prorocentrum minimum belongs to the nucleic acid detection field. It is used for S1 enzyme protection analysis and sandwich hybridization detection of the Prorocentrum minimum. and comprises three correlative oligonucleotide probes. Wherein a S1 enzyme protection analysis probe only serves as a complement of the nucleotides in the Prorocentrum minimum ribosome big subunit 430~520 regions, and shows no homology with other Prorocentrum minimum ribosome RNAs. The S1 enzyme protection analysis probe has a length of 58bp, and can be combined specifically onto the rRNAs of the Prorocentrum minimum under certain conditions, and the sequence is: 5'-ACAGTCCGCAAATGAGTTCTGCCAAGGCTATTCACTCACCCGTAGACGAGCTACCATGA. The invented probes have excellent applicability to make it convenient to seek and realize the specific probe. The invention has higher stability than that of the sandwich hybridization technology.

Description

technical field [0001] The invention relates to a group of oligonucleotide probes for algae detection, in particular to specific probes for qualitative and quantitative detection of Prorocentrum micronella. It belongs to the field of nucleic acid detection. Background technique [0002] Prorocentrum minimum is a common dinoflagellate algae widely distributed in coastal areas, estuaries, oceanic waters, and China's Bohai Sea, East China Sea, South China Sea and other waters. It is one of the main red tide organisms. Red tides have occurred in the waters, causing a large number of dead fish to float on the sea surface. Prorocentrum micronella can produce diarrhea shellfish poisoning and cause human poisoning, which poses a huge threat to micro fishery and human health. Therefore, it is of great significance to timely, accurately and quickly detect the population dynamics of Prorocentrum microbes in microenvironments, and keep abreast of their quantity changes in microenviron...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 李荣秀于志刚蔡青松米铁柱甄毓
Owner SHANGHAI JIAOTONG UNIV
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