Method of qualitative and quantitative analysis for algae

A technology for analyzing algae and probes, which is applied in the field of qualitative and quantitative analysis of algae, and can solve problems such as lack of microalgae, large fluctuations in test results, and difficult operations

Inactive Publication Date: 2007-12-05
OCEAN UNIV OF CHINA +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its main disadvantages are: (a) because the length of the capture probe is only 10-30bp, the number of capture probes that can effectively distinguish close relatives of microalgae is very small; (b) RNA must be prevented 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
[0010] In summary, there is currently a lack of fast, easy and reliable identification methods for microalgae, so there is an urgent need in this field to develop new methods for rapid, simple and reliable identification of the type and / or quantity of microalgae

Method used

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  • Method of qualitative and quantitative analysis for algae
  • Method of qualitative and quantitative analysis for algae
  • Method of qualitative and quantitative analysis for algae

Examples

Experimental program
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Effect test

Embodiment 1

[0070] Example 1: Qualitative and quantitative analysis of Chaetoceros using S1 enzyme protection analysis for rRNA and sandwich hybridization technology

[0071] In this example, the S1 enzyme protection analysis technique and the sandwich hybridization technique are combined to realize the qualitative and quantitative analysis of the marine red tide organism Chaetoceros sp., and the steps are as follows:

[0072] 1.1 Search for the specific region of Chaetoceros rRNA and the design and synthesis of S1 enzyme protection analysis probe

[0073] By determining the 18S rRNA sequence of the ribosomal small subunit of Chaetoceros algae and performing multiple sequence comparisons with the rRNA sequences of other algae, it was found that the 550-700 region of the small subunit rRNA was different from other algae, and the sequence of this region was determined to be a characteristic sequence; The S1 enzyme protection analysis probe CHV_S1 was designed and synthesized according to th...

Embodiment 2

[0104] Example 2: Qualitative and quantitative analysis of Alexandrium tamarensis by using S1 enzyme protection analysis and sandwich hybridization technology for rRNA

[0105] In this embodiment, the S1 enzyme protection analysis technique and the multiple sandwich hybridization technique are combined to realize the qualitative and quantitative analysis of Alexandrium tamarense, and the steps are as follows:

[0106] 2.1 The search for the specific region of Alexandrium tamarina rRNA and the design and synthesis of the S1 enzyme protection analysis probe

[0107] By determining the 18S rRNA sequence of the ribosomal small subunit of Alexandrium tamarica and performing multiple sequence comparisons with the rRNA sequences of other algae, it was found that the 600-800 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 ALT S...

Embodiment 3

[0135] Example 3: Qualitative and quantitative analysis of Chaetoceros using S1 enzyme protection analysis for rRNA and sandwich hybridization technology

[0136] In this example, the steps of Example 1 were repeated, the only difference being that instead of using a universal signal probe, fluorescein was directly labeled at the 5' end of the specific signal probe (that is, connecting the probe and the signal probe The functions of the needle are combined into one).

[0137] As a result, the qualitative results of Chaetoceros completely the same as those in Example 1 were obtained, and the quantitative detection results were almost completely the same. However, the disadvantage of this embodiment is that a specific signal probe with a label needs to be synthesized for each group of detection probes, and the cost of synthesis is relatively high, while the general signal probes used in Examples 1 and 2 can be widely used. Universal, it is not necessary to synthesize a specific...

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Abstract

The invention discloses a method for fast determination of algae species and quantity by employing rRNA-targeted S1 enzyme protection probe and sandwich hybridization technique, which comprises, (1) mixing and hybridizing the detected sample with S1 enzyme protection analyzing probe, (2) treating the mixed solution in step (1) through S1 enzyme digestion, (3) carrying out denaturation treatment to release S1 enzyme protection probe, (4) trapping residual S1 enzyme protection analyzing probes with trapping probes fixed on the solid-phase carriers, (5) hybridizing the linking probe with S1 enzyme protection analyzing probe, then hybridizing with the signal probe with detectable signals, (6) detecting the detectable signals to determine the algae species and / or quantity in the sample.

Description

technical field [0001] The invention relates to the detection field, in particular to a qualitative and quantitative analysis method for algae. More specifically, it relates to a method for quickly identifying the type and quantity of algae by using the S1 enzyme protection analysis probe for rRNA and sandwich hybridization technology. Background technique [0002] Red tide is a kind of harmful ecological abnormal phenomenon in which some microscopic planktonic algae, protozoa or bacteria in the ocean reproduce explosively (proliferate) or gather under certain conditions, causing the water body to change color. From a global perspective, marine planktonic microalgae are the main organisms that cause red tides. Among the more than 4,000 types of marine planktonic microalgae, more than 260 species can form red tides, and more than 70 species of them can produce toxins. When red tides occur, a large number of fish and shellfish die, resulting in major economic losses and serio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/28C12Q1/68
Inventor 于志刚李荣秀蔡青松米铁柱甄毓
Owner OCEAN UNIV OF CHINA
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