Method for detecting content of human enteroviruses and non-human enteroviruses in environmental water

A technology of enterovirus and detection method, which is applied in biochemical equipment and methods, microbiological measurement/inspection, fluorescence/phosphorescence, etc., can solve the problems of large interference, inaccurate quantification, and insufficient sensitivity, and achieve high sensitivity and The effect of precision

Inactive Publication Date: 2013-01-23
XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The hosts of enteroviruses in environmental waters are very different, and the species are complex and low in concentration. Most of the existing PCR detection methods for enteroviruses in environmental waters are qualitative or quantitative detection of total enteroviruses, which are derived from human secretions or excretions. The detection of enteroviruses has rarely been studied
Due to the complex water quality of the environmental water and the presence of many PCR reaction inhibitors and other impurities, the existing detection methods have the disadvantages of insufficient sensitivity, great influence of impurities, and inability to accurately quantify
There are currently no scientifically available methods for the detection of human enteroviruses and non-human enteroviruses in environmental waters

Method used

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  • Method for detecting content of human enteroviruses and non-human enteroviruses in environmental water
  • Method for detecting content of human enteroviruses and non-human enteroviruses in environmental water
  • Method for detecting content of human enteroviruses and non-human enteroviruses in environmental water

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Embodiment

[0055] The equipment and reagents used in this embodiment are as follows:

[0056] (1) High-speed refrigerated centrifuge with a maximum speed of 14000r / min;

[0057] (2) 4℃, -80℃ refrigerator

[0058] (3) Centrifuge tubes, 50mL, 1.5mL, 0.5mL, 0.2mL;

[0059] (4) Trace ultraviolet spectrophotometer;

[0060] (5) Constant temperature metal heater;

[0061] (6) Bacterial shaker, bacterial incubator;

[0062] (7) Real-time fluorescent quantitative PCR instrument;

[0063] (8) Gradient qualitative PCR instrument;

[0064] (9) Electrophoresis apparatus;

[0065] (10) Gel imager;

[0066] (11) pMD19-T vector;

[0067] (12) Competent Escherichia coli DH5α;

[0068] (13) Hydrochloric acid, NaCl solution, MgCl 2 solution, etc.;

[0069] (14) Polyethylene glycol (PEG-6000) with a molecular weight of 6000, glucose, etc.;

[0070] (15) 5mol / L NaCl solution;

[0071] (16) IPTG / X-gal / Amp plate;

[0072] (17) LB liquid medium;

[0073] (18) Total RNA extraction kit;

[0074] ...

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Abstract

The invention discloses a method for detecting content of human enteroviruses and non-human enteroviruses in environmental water. On the basis of the established method for quantitatively detecting the enteroviruses in the environmental water, primers and probes for the human enteroviruses are selected, recombinant plasmids are prepared as standard substances, a real-time fluorescent quantitative PCR reaction system for quantitatively detecting the human enteroviruses is established, the content of the human enteroviruses and the enteroviruses in environmental water is quantitatively detected to further determine a distribution condition of the non-human enteroviruses. The method has good specificity and high sensitivity, accurately quantifies the human enteroviruses and the enteroviruses and determines relationship between the human enteroviruses and the non-human enteroviruses in the environmental water.

Description

technical field [0001] The invention belongs to the technical field of detection of environmental water quality, in particular to a method for detecting the content of human enterovirus and non-human enterovirus in the environment water. Background technique [0002] Enteroviruses are widely present in source water and polluted water, causing outbreaks of water-borne viral infectious diseases. Virus detection methods mainly include: tissue culture method, immunological method, molecular biology method and so on. Among them, the tissue culture method has a long culture cycle, and some viruses lack sensitive cell lines to culture or have no obvious cytopathic effect; immunological detection methods require a higher concentration of virus samples, and it is difficult to reach the required concentration after the environmental water sample is concentrated; Molecular biology methods mostly use the detection of nucleic acids, because of their short detection time, low detection l...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 周进宏王晓昌张崇淼船水尚行佐野大辅
Owner XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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