Escherichia coli O157:H7 and salmonella typhimurium co-testing paper, preparation method and application thereof

A technology of O157 and Escherichia coli, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of poor sensitivity and difficult to overcome cross-reaction, and achieve the effect of strong sensitivity, improved specificity and good specificity

Inactive Publication Date: 2017-10-24
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these test papers generally use colloidal gold particles as signal elements, which makes the sensitivity poor; in addition, the design of single multiple detection lines is used in the detection of multi-target detection, which makes the cross-reaction difficult to overcome

Method used

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  • Escherichia coli O157:H7 and salmonella typhimurium co-testing paper, preparation method and application thereof
  • Escherichia coli O157:H7 and salmonella typhimurium co-testing paper, preparation method and application thereof
  • Escherichia coli O157:H7 and salmonella typhimurium co-testing paper, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Preparation and assembly of Escherichia coli O157:H7 and Salmonella typhimurium co-detection test paper

[0051] 1. Preparation of platinum palladium nanoparticles

[0052] Glassware required for the preparation must be kept in freshly prepared aqua regia (HNO 3 :HCl=3:1) after soaking for 30 minutes, rinse with a large amount of double distilled water and dry in an oven for later use.

[0053] First, configure 20mM potassium tetrachloroplatinate solution (K 2 Ptcl 4 ), 20mM sodium tetrachloropalladate solution (Na 2 Pdcl 4 ), 6M hydrochloric acid solution and 100mM ascorbic acid solution. Then, 20 mg of F127 was weighed and dissolved in 1.8 mL of potassium tetrachloroplatinate solution (20 mM), then 0.2 mL of sodium tetrachloropalladate solution (20 mM) was added, and F127 was completely dissolved by ultrasonication for 5 min. Continue to add 44 μL of Hcl (6M) and 2 mL of ascorbic acid solution (100 mM), and sonicate for 4 hours. After centrifuging at ...

experiment example 3

[0068] Experimental example 3 utilizes the test paper prepared in embodiment 1 to carry out the detection of actual samples

[0069] In order to further verify the applicability of the sensor, a spike recovery experiment was carried out. Add 10 to 2.5mL milk and ice cream respectively 4 、10 5 CFU mL -1 coli O157:H7 and Salmonella typhimurium, mix for 1 min. Then, the sensor technology was used to quantitatively detect and analyze Escherichia coli O157:H7 and Salmonella typhimurium. The results showed (Table 1) that the recovery rate was between 91.44% and 109.56%, which proved that the sensor could be used for on-site detection.

[0070] Table 1 Recovery rate of Salmonella in artificially contaminated milk and ice cream

[0071]

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Abstract

The invention relates to a co-detection test paper for Escherichia coli O157:H7 and Salmonella typhimurium, comprising a base plate; a sample pad attached to the base plate; two absorbent strips arranged in parallel on the base plate and not in contact with each other; The goat-derived Salmonella typhimurium monoclonal antibody and the goat-derived Escherichia coli O157:H7 monoclonal antibody are used as the detection line respectively, and the goat anti-mouse IgG antibody is used as the quality control line; the binding pad is coated with platinum-palladium nanoparticles-mouse-derived Typhimurium Salmonella monoclonal antibody complex, platinum-palladium nanoparticles-mouse E. coli O157:H7 monoclonal antibody complex. The invention uses platinum palladium nanoparticles instead of traditional colloidal gold as a signal element to improve sensitivity, and uses a parallel dual-channel design instead of a single multiple detection line design to improve specificity, and can be used for simultaneous detection of Escherichia coli O157:H7 and Salmonella typhimurium.

Description

technical field [0001] The invention belongs to the field of detection of food-borne pathogenic microorganisms in food safety, and relates to a method for preparing immunochromatographic test paper based on nanozyme, using platinum palladium nanoparticles instead of traditional colloidal gold as a signal element to improve sensitivity, and using parallel double Channel design replaces single multi-detection line design to improve specificity, and establishes a biosensor detection technology with strong sensitivity and specificity, which is used for rapid on-site detection of common foodborne pathogenic bacteria Escherichia coli O157:H7 and Salmonella typhimurium detection. Background technique [0002] Nanozyme is a signal element with good performance in the construction of biosensors discovered in recent years. It is named for the catalytic activity of this type of nanoparticle. Generally, nanozyme-based biosensors can improve sensitivity and lower the minimum detection l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/577G01N33/56916
Inventor 罗云波许文涛程楠黄昆仑徐瑗聪杨湛森贺晓云
Owner CHINA AGRI UNIV
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