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Real-time fluorescent quantitative PCR detection method for phosphorus accumulating bacteria in sewage treatment plants

A real-time fluorescence quantitative, sewage treatment plant technology, used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve high amplification efficiency, good linearity, and good versatility.

Active Publication Date: 2014-10-15
BEIJING UNIV OF TECH
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Problems solved by technology

The established standard curve was used to quantitatively detect the phosphorus-accumulating bacteria in the whole-process denitrification and phosphorus-removal system and the short-term denitrification and phosphorus-removal system of sewage, and the population structure of phosphorus-accumulating bacteria and the effect of gene expression of polyphosphate kinase on the actual sewage were established by changing the operating conditions. There are no relevant research reports on the dynamic response mechanism of environmental condition changes in nitrogen and phosphorus removal systems

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  • Real-time fluorescent quantitative PCR detection method for phosphorus accumulating bacteria in sewage treatment plants
  • Real-time fluorescent quantitative PCR detection method for phosphorus accumulating bacteria in sewage treatment plants
  • Real-time fluorescent quantitative PCR detection method for phosphorus accumulating bacteria in sewage treatment plants

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Embodiment Construction

[0045] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0046] 1.PCR primers

[0047] Using polymeric phosphate kinase gene as a marker, PCR primers for five evolutionary branches of phosphorus accumulating bacteria were selected. The primer names, primer sequences, and amplicon lengths are shown in Table 2.

[0048] Table 2 Primer sequences used in conventional PCR amplification

[0049]

[0050] 2. The source of the standard

[0051] The sources of standard products used in the present invention are all reactors for treating actual domestic sewage. The influent in the anaerobic zone of the reactor has two parts, the test raw water and the reflux liquid in the anoxic zone 1, and the NO in the two parts of the influent 2 - -N and NO 3 - -N content is lower, eliminating NO 2 - -N and NO 3 - The effect of -N on the anaerobic phosphorus release reaction enables the phosphorus accumulating bacteria...

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Abstract

The invention provides a sewage treatment plant phosphorus-accumulating bacterium real-time fluorescence quantification polymerase chain reaction (PCR) detection method, and belongs to the technical field of biological sewage treatment. According to the method, phosphorus-accumulating bacteria enriched in actual life sewage serve as a source of standard substances, and flora evolutionary branches are rich. Aiming at key function genes of phosphorus-accumulating bacteria, specific primers of polymerization phosphate kinase genes are adopted, standard substances of I, IIA, IIB, IIC and IID belonging to five main evolutionary branches are obtained, and SYBR Green fluorescent dye method which is good in generality and low in cost is adopted to set up real-time fluorescence quantification PCR standard curves of the five main evolutionary branches. Correlation coefficients of the five standard curves are respectively 1.000, 0.998, 0.999, 0.999 and 0.998. The amplification efficiency is between 90% and 110%. The established standard curves are used for carrying out quantitative detection for different evolutionary branches of the phosphorus-accumulating bacteria in a certain sewage treatment system. The method is economical, simple, convenient, and reliable.

Description

technical field [0001] The invention relates to a real-time fluorescent quantitative PCR detection method for phosphorus-accumulating bacteria in sewage treatment plants, which belongs to the technical field of sewage biological treatment and is used for quantitative detection of phosphorus-accumulating bacteria in sewage biological treatment systems. Background technique [0002] Phosphorus is a key nutrient element leading to eutrophication of surface water bodies. All countries in the world have established strict discharge standards for the concentration of phosphorus discharged into surface water bodies. Biological phosphorus removal technology has been widely used in sewage treatment plants in major cities due to its cost-effective features. Although biological phosphorus removal technology has been applied to sewage treatment for decades and has accumulated a lot of operating experience, unexplained deterioration of phosphorus removal effect often occurs during the op...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 曾薇李博晓张立东王向东彭永臻
Owner BEIJING UNIV OF TECH
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