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Salmonella enrichment and rapid detection method

A Salmonella detection method technology, applied in the field of microbial detection, can solve the problems of sensitivity limitation, affecting the results and sensitivity, etc., and achieve the effect of high coupling rate, small variation coefficient and uniform immunological reaction

Active Publication Date: 2013-12-11
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for some samples with extremely low antigen or antibody content, the color of the marker is so light that it is almost difficult to judge the result with the naked eye. The instrument can only detect the color of the marker on the surface of the solid phase carrier, but it is difficult to detect the marker inside the solid phase carrier, so the detection sensitivity is greatly limited
The biological samples detected at the same time may contain color substances, which will seriously affect the results and sensitivity of the detection

Method used

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  • Salmonella enrichment and rapid detection method
  • Salmonella enrichment and rapid detection method
  • Salmonella enrichment and rapid detection method

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Example 1: Detection of Salmonella in milk using nano-gold magnetic particles

[0028] 1. Preparation of gold magnetic particles coupled with monoclonal antibody:

[0029] 1.1 Treatment of nano-gold magnetic particles: Take 200-400 μL of coupling buffer in a 2 mL centrifuge tube, mix with 0.5-1.0 mg of 50 nm nano-gold magnetic particles, magnetically separate for 3-5 minutes, and discard the supernatant.

[0030] 1.2 Coupling reaction: Take 200-300 μg of the prepared anti-Salmonella monoclonal antibody, mix with 0.5-1.0 mg of 50 nm nano-gold magnetic particles, and place in 1 mL of coupling buffer. At a temperature of 37°C, place on a rotator with a rotational speed of 10-15 rpm, couple for 30-60 min, magnetically separate for 3-5 min and discard the supernatant. Wash 3 times with 1 mL of wash buffer.

[0031] 1.3 Blocking: After washing, mix 1 mL of blocking agent with magnetic beads to block for 1 h.

[0032] 2. Capturing Salmonella in milk using nano-gold magnetic...

Embodiment 2

[0040] Example 2: Detection of Salmonella in beef using nano-gold magnetic particles

[0041] 1. Preparation of gold magnetic particles coupled with monoclonal antibody:

[0042] 1.1 Treatment of nano-gold magnetic particles: Take 200-400 μL of coupling buffer in a 2 mL centrifuge tube, mix with 0.5-1.0 mg of 50 nm nano-gold magnetic particles, magnetically separate for 3-5 minutes, and discard the supernatant.

[0043] 1.2 Coupling reaction: Take 200-300 μg of the prepared anti-Salmonella monoclonal antibody, mix with 0.5-1.0 mg of 50 nm nano-gold magnetic particles, and place in 1 mL of coupling buffer. At a temperature of 37°C, place on a rotator with a rotational speed of 10-15 rpm, couple for 30-60 min, magnetically separate for 3-5 min and discard the supernatant. Wash 3 times with wash buffer.

[0044] 1.3 Blocking: After washing, mix 1 mL of blocking agent with magnetic beads to block for 1 h.

[0045] 2. Capturing Salmonella in milk using nano-gold magnetic parti...

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Abstract

The invention adopts double-function gold magnetic nanoparticles and provides a detection method which integrates an immunomagnetic bead capturing technology and an immunochromatography technology and is used for rapidly detecting salmonellas. Immunomagnetic separation and immunochromatography are organically integrated, the step of eluting the salmonellas from immunomagnetic beads is eliminated and the capture efficiency is improved; the step of spraying colloidal gold on a bonding mat is eliminated, the immunological reaction is more uniform and a variable coefficient is small in the quantitative detection; workload and probability of mixed bacterium pollution are reduced. The detection method adopts the basic thinking of exploring the mode of effectively combining the gold magnetic nanoparticles and an antibody, optimizing the conditions of enriching the salmonellas and carrying out rapid quantitative detection by using the immunochromatography technology as a vector and using the double antibodies sandwich as a detection principle.

Description

technical field [0001] The invention relates to the field of microorganism detection, and specifically adopts a bifunctional nano-gold magnetic particle integrated immune magnetic bead capture technology and immune chromatography to rapidly detect Salmonella. technical background [0002] Salmonella is a kind of food-borne pathogenic bacteria ubiquitous in nature. Among the food poisoning cases in the world, the poisoning cases caused by Salmonella occupy the first place. They often cause diseases such as acute diarrhea, vomiting, abdominal pain, high fever and sepsis in humans. Salmonella is more harmful. In the 2006 EU investigation, about 160,049 people were confirmed to be infected with Salmonella. From April to August 2008, 286 people were infected and 2 died in the United States. In 2010, about 500 million contaminated eggs were recalled in the United States due to salmonella contamination. In 2010, 203 surveys were sampled from 4090 farms in Japan, and 48 farms show...

Claims

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Application Information

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IPC IPC(8): G01N33/569
Inventor 赖卫华山珊
Owner NANCHANG UNIV
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