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Detection probe for Heterosigma akashiwo Hada

A technology for detecting H. akashiwo and detection probes, which is applied in the measurement/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of large fluctuations in detection results, small number of captured probes, and difficult operation, and achieve Easy to find and realize, good probe applicability and high stability

Inactive Publication Date: 2005-07-06
SHANGHAI JIAO TONG 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 Heterosigma akashiwo Hada
  • Detection probe for Heterosigma akashiwo Hada

Examples

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Embodiment

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

[0031] In this example, the S1 enzyme protection analysis technique and the sandwich hybridization technique are combined to realize the qualitative and quantitative detection of the marine red tide organism Heterosigma akashiwo, and the steps are as follows:

[0032] 1.1 Search for specific regions of H. akashiwo rRNA and design and synthesis of probes for S1 enzyme protection analysis

[0033] By determining the 18S rRNA sequence of the ribosomal small subunit of Heterocurvium akashiwo and performing multiple sequence comparisons with the rRNA sequences of other algae, it was found that the 150-300 region of the small subunit rRNA was different from other algae, and the sequence of this region was determined to be a characteristic sequence ; Design and synthesize the S1 enzyme protection analysis probe HER_S1 ac...

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Abstract

A detection probe for Heterosigma akashiwo belongs to the nucleic acid detection field. It is used for S1 enzyme protection analysis and sandwich hybridization detection of the Heterosigma akashiwo, and comprises three correlative oligonucleotide probes. Wherein a S1 enzyme protection analysis probe only serves as a complement of the nucleotides in the Heterosigma akashiwo ribosome small subunit 150~300 regions, and shows no homology with other Heterosigma akashiwo ribosome RNAs. The S1 enzyme protection analysis probe has a length of 59bp, and can be combined specifically onto the rRNAs of the Heterosigma akashiwo under certain conditions, and the sequence is: 5'-GGCAGAAACTTGAATGAACCATCGACCGAAGTCGATTCGCACAGTTACTATGATTCACC. 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 Heterocurvium akashiwo. It belongs to the field of nucleic acid detection. Background technique [0002] Heterosigma akashiwo is a flagellar algae that is widely distributed in coastal waters around the world. It is one of the main red tide organisms. It has repeatedly formed harmful red tides and caused a large number of deaths of farmed fish. pose a serious threat to human health. Therefore, it is of great significance to timely, accurately and quickly detect the population dynamics of H. akashiwo in the marine environment and keep abreast of its quantity changes in the ocean. The traditional identification method of directly observing and counting with an optical microscope is cumbersome, time-consuming, and laborious, and requires the participation of experienced taxonomy experts. When the...

Claims

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Application Information

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