Fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from sewage treatment plant

A sewage treatment plant, nitrite technology, applied in biochemical equipment and methods, fluorescence/phosphorescence, microorganism-based methods, etc., to achieve guaranteed effects, rich flora, and strong applicability.

Active Publication Date: 2014-02-05
BEIJING UNIV OF TECH
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Problems solved by technology

Use the established standard curve to quantitatively detect the nitrite oxidizing bacteria in the whole-process denitrification and phosphorus removal and short-range denitrification and phosphorus removal systems of sewage, and analyze the relationship between the dynam

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  • Fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from sewage treatment plant
  • Fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from sewage treatment plant
  • Fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from sewage treatment plant

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Embodiment Construction

[0033] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0034] 1. Establishment of real-time fluorescence quantitative PCR standard curve

[0035] 1.1PCR primers

[0036] The primer sequences and amplicon lengths of the two genera of nitrite oxidizing bacteria, Nitrobacter and Nitrospira, are shown in Table 2.

[0037] Primer sequences used in table 2 PCR amplification

[0038]

[0039] 1.2 Extraction of total DNA

[0040] The activated sludge was taken from the aerobic zone of the continuous flow reactor for treating actual domestic sewage, and the genomic DNA of activated sludge microorganisms was extracted using the DNA extraction kit (Fast DNA Spin kit for soil, BIO101system, USA) produced by MPBiomedicals, USA. The operation is carried out in strict accordance with the instructions. 1.3 Conventional PCR

[0041] The primers used in the routine PCR of Nitrobacter and Nitrospira are shown in Table...

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Abstract

A real-time fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from a sewage treatment plant belongs to the technical field of biological sewage treatment. According to the invention, two genuses of nitrite oxidizing bacteria from a sewage treatment plant, namely nitrobacter and nitrospira, can be detected. All nitrite oxidizing bacteria can be analyzed, and dynamic changes of each genus can be analyzed. By a SYBR Green fluorochrome method with good versatility and low cost, real-time fluorescent quantitative standard curves of the two genuses of nitrite oxidizing bacteria are established. Correlation coefficients of the two standard curves are respectively 0.998 and 0.997, and amplification efficiency is between 100% and 110%. The established standard curves are utilized to monitor and identify nitrite oxidizing bacteria in a sewage treatment system. The invention provides an economical, simple and reliable real-time fluorescent quantitative PCR detection method of nitrite oxidizing bacteria from a sewage treatment plant.

Description

technical field [0001] The invention relates to a real-time fluorescence quantitative PCR detection method for nitrite oxidizing bacteria in sewage treatment plants, belongs to the technical field of sewage biological treatment, and is used for quantitative detection of nitrite oxidizing bacteria in sewage biological treatment systems. Background technique [0002] Sewage biological denitrification technology is widely used in the field of sewage treatment due to its economical and high-efficiency characteristics. The principle of traditional sewage biological denitrification is that ammonia nitrogen is oxidized by ammonia oxidizing bacteria to nitrite nitrogen (nitrosation stage), and then oxidized by nitrite oxidizing bacteria to nitrate nitrogen (nitration process); after that, nitrate nitrogen and nitrite The state nitrogen is denitrified into nitrogen gas (denitrification process), thereby realizing the removal of nitrogen in sewage. For the traditional biological deni...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64C12R1/01
CPCC12Q1/04C12Q1/6851C12Q1/686C12Q2545/114C12Q2561/113C12Q2531/113
Inventor 曾薇白鑫龙张丽敏彭永臻
Owner BEIJING UNIV OF TECH
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