Method for sensitively, simply and conveniently detecting bacteria

A technology of bacteria and targets, applied in the field of microbiology and chemical biology, can solve the problems of many steps and complicated detection process, and achieve the effect of simple steps, simple detection process and high sensitivity

Inactive Publication Date: 2012-12-26
WUHAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in most of these detection methods, capture and labeling are carried out separately, there are many steps, and the detection process is relatively cumbersome.

Method used

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  • Method for sensitively, simply and conveniently detecting bacteria
  • Method for sensitively, simply and conveniently detecting bacteria
  • Method for sensitively, simply and conveniently detecting bacteria

Examples

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Embodiment 1

[0026] Example 1 Salmonella typhimurium S . typhimurium detection

[0027] Salmonella typhimurium S . typhimurium As an example, the method of the present invention is described in detail.

[0028] 1. Method

[0029] 1. Preparation of immunomagnetic and immunofluorescent spheres

[0030] The preparation of magnetic nanospheres (MNS) and fluorescent nanospheres (FNS) is based on the preparation of fluorescent magnetic bifunctional nanospheres in our laboratory. That is, the ultrasonic swelling method is used to make the oil-soluble magnetic nanoparticles γ-Fe 2 o 3 Alternatively, quantum dots (CdSe / ZnS QDs) were embedded into the hydrophobic cavity of surface carboxylated styrene-acrylamide copolymer nanospheres (Pst-AAm-COOH), resulting in MNS and FNS. The diameter of the nanospheres is about 250 nm. Before attaching the antibody on the surface of the nanosphere, introduce NH by covalent coupling 2 -PEG-COOH (MW 3400) as a bridge (spacer): Take about 5 mg MNS or...

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Abstract

The invention relates to a method for detecting bacteria. The method comprises the steps as follows: coupling magnetic nanospheres and fluorescent nanospheres with mouse typhus salmonella antibodies so as to obtain mouse typhus salmonella-targeted immunologic magnetic spheres and immunologic fluorescent spheres, and adding the mouse typhus salmonella-targeted immunologic magnetic spheres and immunologic fluorescent spheres into a detection system so as to realize magnetic capture and fluorescent labeling to the mouse typhus salmonella simultaneously. About 10 CFU/mL of the mouse typhus salmonella can be detected through observation of a fluorescent microscope; and a good quantitative relation is obtained through detection of a fluorescence spectrophotometer, the linear range is 105-107 CFU/mL, and R2 is equal to 0.9994. The whole detection process is very simple, high in sensitivity and strong in specificity, and can be finished detection within 1.5 h.

Description

[0001] technical field [0002] The invention relates to a method for detecting bacteria, belonging to the fields of microbiology and chemical biology. Background technique [0003] Food-borne pathogenic bacteria seriously threaten human health and life. Many people get sick, hospitalized and even die every year, and because the infectious dose of many pathogenic bacteria is very low (for example, the infectious dose of enterohaemorrhagic E. coli O157:H7 and Salmonella As low as 10 cells), so simple, fast, sensitive and reliable detection of bacteria is very important for human life and health. However, traditional detection methods, such as culture methods and enzyme-linked immunosorbent assay (ELISA), have disadvantages such as time-consuming and low sensitivity, while PCR methods, although highly sensitive, usually require sample pretreatment and professional operations and steps cumbersome, which limits its application. [0004] In recent years, with the rapid developm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/533G01N21/64
Inventor 庞代文温聪颖胡军张志凌孙自镛
Owner WUHAN UNIV
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