Detection method for CVA16 in environmental water body

A detection method and virus technology, applied in the directions of microorganism-based methods, microbial determination/inspection, biochemical equipment and methods, etc., which can solve the problems of complex factors affecting detection, inability to accurately qualitatively or quantitatively detect CVA16, and no CVA16 detection method, etc. question

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

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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 CVA16 in the environmental water body is low, and the factors affecting

Method used

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  • Detection method for CVA16 in environmental water body
  • Detection method for CVA16 in environmental water body
  • Detection method for CVA16 in environmental water body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0074] The equipment and reagents used in this example are as follows:

[0075] (1) High-speed refrigerated centrifuge, the maximum speed is 14000r / min;

[0076] (2) 4°C, -80°C refrigerator

[0077] (3) Centrifuge tubes, 50mL, 1.5mL, 0.5mL, 0.2mL;

[0078] (4) Micro UV spectrophotometer;

[0079] (5) Constant temperature metal heater

[0080] (6) Bacterial shaker, bacterial incubator

[0081] (7) Real-time fluorescence quantitative PCR instrument;

[0082] (8) Gradient qualitative PCR instrument;

[0083] (9) Electrophoresis apparatus;

[0084] (10) Gel imager

[0085] (11) pMD19-T vector;

[0086] (12) Competent Escherichia coli DH5α:

[0087] (13) Hydrochloric acid, NaCl solution, MgCl 2 solution, etc.;

[0088] (14) Polyethylene glycol (PEG-6000) with a molecular weight of 6000, glucose, etc.;

[0089] (15) 5mol / L NaCl solution;

[0090] (16) IPTG / X-gal / Amp plate;

[0091] (17) LB liquid medium;

[0092] (20) Total RNA extraction kit;

[0093] (18) Reverse t...

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Abstract

The invention discloses a detection method for CVA16 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 CVA16 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 CVA16 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 CVA16 virus in the environmental water body accurately.

Description

technical field [0001] The invention belongs to the technical field of detection of environmental water quality, in particular to a detection method for CVA16 virus in environmental water. technical background [0002] Coxsackievirus A16 (CVA16) is one of the main pathogens causing hand, foot and mouth disease and belongs to the genus Enterovirus. CVA16 is a water-borne virus that can leak into environmental water from the secretions of CVA16-infected patients, and can survive in environmental water for a long time. The water bodies polluted by it become an important source of CVA16 pollution, which can lead to the outbreak of hand, foot and mouth disease. [0003] The existing methods for detecting CVA16 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 microorganisms interfere with the detection of CVA16 in cell c...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
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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