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Thiobacillus ferrooxidans detection probe composition and related detection method

A technology of Thiobacillus ferrooxidans and detection probe is applied in the field of leaching bacteria detection and bacteria detection, which can solve the problems of influence of leaching efficiency, difficult repeatability, extreme operating conditions, etc. The effect of improving detection accuracy and improving stability

Active Publication Date: 2012-12-26
JIANGNAN UNIV
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Problems solved by technology

Because in the large-scale bioleaching site, the leaching effect is a mixture of various bacteria, and the change of the condition control in different stages of leaching will lead to the change of the bacterial flora, which will inevitably affect the leaching efficiency.
However, at this stage, there are relatively few studies on the changes of bioleaching bacterial flora, and the research methods are limited to microscopes and PCR techniques. Each method has certain advantages and limitations. Operators have high requirements, and in the leaching process, it is difficult to exclude strains with close kinship, and the qualitative effect is not good; timed quantitative PCR technology is expensive, has high requirements for DNA purification efficiency, and is difficult to repeat Wait
[0003] Although there have been reports in China in recent years that the improved sandwich hybridization method has been used for qualitative and quantitative detection of algae, due to the turbidity of the leaching sample liquid, extreme operating conditions and very simple conditions in the bioleaching site, there is no efficient detection of Thiobacillus ferrooxidans. method

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  • Thiobacillus ferrooxidans detection probe composition and related detection method
  • Thiobacillus ferrooxidans detection probe composition and related detection method
  • Thiobacillus ferrooxidans detection probe composition and related detection method

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Embodiment Construction

[0030] In order to understand the technical content of the present invention more clearly, the following examples are given in detail for the purpose of better understanding the content of the present invention rather than limiting the protection scope of the present invention. In this example, specific probes were used to detect Thiobacillus ferrooxidans, the main bacterial species of bioleaching, to prepare its standard curve, and to verify its specificity.

[0031] 1.1 Design and synthesis of probes for specific regions of Thiobacillus ferrooxidans 16S rRNA

[0032] Comparing the 16S rRNA sequence of Thiobacillus ferrooxidans with other ore-leaching bacteria sequences, the nucleic acid complementation in the 896-965 region was found, but there was no homology with other main ore-leaching bacteria nucleotides, and it was determined as a characteristic region, and the analysis was designed The probes are:

[0033] 5'-ATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTCCGGAAT...

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Abstract

The invention relates to a thiobacillus ferrooxidans detection probe composition comprising an analysis probe used to be specifically combined with the rRNA of the thiobacillus ferrooxidans, a capturing probe used to be combined on a carrier and is used to be specifically combined with the analysis probe, and a signal probe used to be specifically combined with the analysis probe. The signal probe has protein markers. Preferably, the analysis probe has a nucleotide sequence represented by SEQ ID No. 1, the capturing probe has a nucleotide sequence represented by SEQ ID No.2 and has biotin markers, and the signal probe has a nucleotide sequence represented by SEQ ID No. 3. The protein markers are fluorescein markers. The invention also provides a thiobacillus ferrooxidans detection method. The thiobacillus ferrooxidans detection probe composition provided by the invention is inelegantly designed. With the detection probe composition, thiobacillus ferrooxidans can be rapidly quantitatively and qualitatively detected. The detection method has the advantages of simple operation and good repeatability. The method is suitable for large-scale popularization.

Description

technical field [0001] The invention relates to the technical field of bacteria detection, more specifically, to the technical field of ore leaching bacteria detection, in particular to a detection probe composition for Thiobacillus ferrooxidans and related detection methods. Background technique [0002] Thiobacillus ferrooxidans is an acidophilic thiobacillus, which belongs to extremophile microorganisms. In most leaching sites, the main strains play a major role in bacterial leaching. Because in the large-scale bioleaching site, the leaching effect is a mixture of various bacteria, and the change of the condition control in different stages of leaching will lead to the change of the bacterial flora, which will inevitably affect the leaching efficiency. However, at this stage, there are relatively few studies on the changes of bioleaching bacterial flora, and the research methods are limited to microscopes and PCR techniques. Each method has certain advantages and limitati...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 杨海麟辛瑜王武冯守帅夏小乐张玲
Owner JIANGNAN UNIV
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