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Detection method for EV71virus in environmental water body

A detection method and virus technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, fluorescence/phosphorescence, etc., and can solve problems such as low EV71 concentration, no EV71 detection method, and inability to accurately qualitatively or quantitatively detect EV71.

Inactive Publication Date: 2013-09-18
XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing PCR detection technology is mainly used in the medical field, and the detection samples basically come from human secretions, while the concentration of EV71 in the environmental water body is low, and the factors affecting the detection are complicated. Quantitative detection
There is currently no suitable method for the detection of EV71 in environmental water

Method used

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  • Detection method for EV71virus in environmental water body
  • Detection method for EV71virus in environmental water body
  • Detection method for EV71virus in environmental water body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0075] The equipment and reagents used in this embodiment are as follows:

[0076] (1) High-speed refrigerated centrifuge with a maximum speed of 14000r / min;

[0077] (2) 4℃, -80℃ refrigerator

[0078] (3) Centrifuge tubes, 50mL, 1.5mL, 0.5mL, 0.2mL;

[0079] (4) Trace ultraviolet spectrophotometer;

[0080] (5) Constant temperature metal heater

[0081] (6) Bacterial shaker, bacterial incubator

[0082] (7) Real-time fluorescent quantitative PCR instrument;

[0083] (8) Gradient qualitative PCR instrument;

[0084] (9) Electrophoresis apparatus;

[0085] (10) Gel Imager

[0086] (11) pMD19-T vector;

[0087] (12) Competent Escherichia coli DH5α:

[0088] (13) Hydrochloric acid, NaCl solution, MgCl 2 solution, etc.;

[0089] (14) Polyethylene glycol (PEG-6000) with a molecular weight of 6000, glucose, etc.;

[0090] (15) 5mol / L NaCl solution;

[0091] (16) IPTG / X-gal / Amp plate;

[0092] (17) LB liquid medium;

[0093] (20) Total RNA extraction kit;

[0094] (18...

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Abstract

The invention discloses a detection method for EV71 virus in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect EV71 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at EV71 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect EV71 virus in the environmental water body accurately.

Description

technical field [0001] The invention belongs to the technical field of detection of environmental water quality, and in particular relates to a detection method of EV71 virus in environmental water. technical background [0002] Enterovirus 71 (EV71), which belongs to the Enterovirus genus, has attracted much attention in recent years and is a major pathogen that causes hand, foot and mouth disease. EV71 is a water-borne virus that can be leaked from the secretions of EV71-infected patients into environmental water bodies and can survive in environmental water bodies for a long time. The water body polluted by it becomes an important pollution source of EV71, which can cause the outbreak of hand, foot and mouth disease. [0003] Existing methods for detecting EV71 mainly include cell culture method and polymerase chain reaction (Polymerase Chain Reaction, PCR). There are a variety of pathogenic microorganisms in environmental water, especially other enteroviruses. These mi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
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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