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Visual detection method for mouse typhus salmonella based on aptamer recognition

A Salmonella, detection method technology, applied in the field of molecular biology and microbiology, to achieve the effect of good linearity, high specificity and easy operation

Inactive Publication Date: 2013-04-03
JIANGNAN UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these rapid detection methods have some defects in detection sensitivity, detection time and specificity.

Method used

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  • Visual detection method for mouse typhus salmonella based on aptamer recognition
  • Visual detection method for mouse typhus salmonella based on aptamer recognition
  • Visual detection method for mouse typhus salmonella based on aptamer recognition

Examples

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Embodiment 1

[0017] Example 1: Application of the visual detection method of Salmonella typhimurium based on aptamer recognition-gold-labeled silver staining to detect water samples

[0018] 1. Synthesis of biotin and thiol-modified Salmonella typhimurium aptamer (completed by Shanghai Sangon Bioengineering Co., Ltd.).

[0019] Biotin-modified aptamer: 5'-biotin-C6-TAT GGC GGC GTC ACC CGA CGG GGA CTT GAC ATTATG ACA G-3'.

[0020] Sulfhydryl-modified aptamer: 5'-SH-TAT GGC GGC GTC ACC CGA CGG GGA CTT GAC ATT ATGACA G-3'.

[0021] 2. Preparation of nano gold.

[0022] Soak the relevant container with aqua regia (concentrated hydrochloric acid and concentrated nitric acid with a volume ratio of 3:1), then clean it with ultrapure water, and dry it. Utilize the method for sodium citrate to reduce chloroauric acid to prepare nano gold:

[0023] 1) Add 99.16mL ultrapure water (≥18.2Ω) and 0.84mL 2.5% chloroauric acid solution into the container, stir evenly with a rotor, and boil for 10min. 2...

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Abstract

The invention provides a visual detection method for mouse typhus salmonella based on aptamer recognition. The basic principle of the method is that the surface of a microwell plate is coated with avidin, sulfhydrylation salmonella aptamer which is marked by biotinylation salmonella aptamer and nanogold is adopted to recognize targeted salmonella, and on the basis, a silver enhancement technique is utilized to further amplify a detection signal, so that the visual detection is achieved. The visual detection method for the mouse typhus salmonella based on the aptamer recognition has the advantages of high specificity, strong sensitivity, simplification and rapidness, thereby providing the possibility for rapidly detecting the mouse typhus salmonella in an environment.

Description

technical field [0001] The present invention mainly realizes the visual detection through the specific recognition of the Salmonella typhimurium aptamer to the Salmonella and the gold-labeled silver staining method. The invention relates to methods and techniques of molecular biology and microbiology, and belongs to the field of biotechnology. Background technique [0002] Salmonella is a facultative anaerobic Gram-negative bacterium belonging to the Enterobacteriaceae family and one of the most important pathogenic genera in the Enterobacteriaceae family. Salmonellosis caused by Salmonella is one of the important zoonotic diseases in public health. In addition, Salmonella can infect many kinds of food. Salmonella is the most common cause of foodborne outbreaks in Europe, the EFSA concluded in its report. Among bacterial food poisoning in my country, 70% to 80% are caused by Salmonella. According to the report of the Centers for Disease Control and Prevention of the Unite...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/10C12Q1/06
CPCY02A50/30
Inventor 王周平袁京磊俞晔段诺吴世嘉夏雨马小媛
Owner JIANGNAN UNIV
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