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A set of probe compositions for the detection of pseudomonas (pseudomonas) bacteria

A technology of pseudomonad and composition, which is applied in the field of probe composition, can solve the problems of high price and difficult repeatability, and achieve the effects of low equipment requirements, short detection time, and low cost of materials and medicines

Active Publication Date: 2020-04-17
JIANGNAN UNIV
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  • Abstract
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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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  • A set of probe compositions for the detection of pseudomonas (pseudomonas) bacteria
  • A set of probe compositions for the detection of pseudomonas (pseudomonas) bacteria
  • A set of probe compositions for the detection of pseudomonas (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'-CGGCTAGTTG ACATCGTTTA CGGCGTGGAC TACCAGGGTA TCTAATCCT...

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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 present invention relates to a set of methods for detecting Pseudomonas ( Pseudomonas ) Bacterial probe composition, specifically relates to DNA-RNA hybridization and DNA-DNA hybridization nucleic acid hybridization using a group of molecular probes, belonging 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...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6834C12Q1/06C12Q1/04C12N15/11
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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