Probe composition group for detecting Pseudomonas bacteria

A technology of pseudomonad and composition, applied in the field of probe composition, can solve the problems of difficult repeatability and high price, and achieve the effect of low equipment requirements, low price and improved stability

Active Publication Date: 2016-07-06
JIANGNAN UNIV
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  • Description
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Problems solved by technology

However, timed quantitative PCR technology and DGGE-PCR technology are expensive, have high requirements for DNA purification efficiency, and are difficult to repeat.

Method used

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  • Probe composition group for detecting Pseudomonas bacteria
  • Probe composition group for detecting Pseudomonas bacteria

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Embodiment Construction

[0028] 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 nitrogen and phosphorus removal bacteria in urban sewage treatment, and their standard curves were prepared to verify their specificity.

[0029] 1. Design and synthesis of probes for specific regions of 16rRNA in Pseudomonas bacteria:

[0030] Comparing the 16rRNA sequence of Pseudomonas bacteria with other sewage bacteria sequences, it is found that it can be complementary to all species of Pseudomonas genus, but has no homology with other main sewage bacteria nucleotides, and it is determined as a characteristic region, and the design The analysis probes are:

[0031] SEQ ID NO: 1:

[0032] 5'-CGGCTAGTTGACATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTT...

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Abstract

A probe composition group for detecting Pseudomonas bacteria belongs to the technical field of nucleic acid detection. The probe composition is composed of three oligonucleotide probes. An analysis probe has as a nucleotide sequence shown in SEQ ID NO:1; a capture probe has a nucleotide sequence shown in SEQ ID NO:2, and has biotin label at the 3' terminal; a signal probe has a nucleotide sequence shown in SEQ ID NO: 3, and has isothiocyanic acid label (FITC) at the 5' terminal. The probe composition has bioinformatics specificity, is able to distinguish Pseudomonas, Comamonas, Aeromonas, Acinetobacter, Bacillus subtilis and Escherichia coli; the probe has quantitative linear range of 7.08*10<3> -6.31*10<6> cells / mL, and the linear equations are as below: y=0.373x-0.962, R<2>=0.996. The probe composition can be used for qualitative and quantitative analysis of Pseudomonas bacteria.

Description

technical field [0001] The invention relates to a set of probe compositions for detecting Pseudomonas bacteria, in particular to nucleic acid hybridization using a set of molecular probes for DNA-RNA hybridization and DNA-DNA hybridization, and belongs to the technical field of nucleic acid detection. Background technique [0002] Pseudomonas bacteria are ubiquitous in soil and water environments and are a class of Gram-negative abacillus, some of which are conditionally pathogenic. Existing research results show that during the biological treatment of sewage, certain species of Pseudomonas can consume nitrogen and phosphorus compounds in sewage, reduce the nitrogen and phosphorus content of water, and purify sewage. Due to the complexity of the sewage system and the coexistence of various microorganisms, how to determine the content of nitrogen and phosphorus removal bacteria in the treatment system is particularly important. [0003] At present, the research on nitrogen a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/06C12Q1/04C12N15/11
Inventor 杨海麟王武辛瑜夏小乐张玲杨继伟梁洪杰
Owner JIANGNAN UNIV
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