Salmonella cleia detection kit and detection method

A detection kit and technology for Salmonella, applied in measurement devices, immunoassays, instruments, etc., can solve the problems of difficult measurement, low luminescence intensity, etc., and achieve the effects of simple operation, improved sensitivity, wide application prospect and development value

Inactive Publication Date: 2016-01-20
徐静 +5
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This traditional chemiluminescent system (HRP-hydrogen peroxide-luminol) flashes instantaneously within a few seconds, and has disadvantages such as low luminous intensity and difficult measurement

Method used

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  • Salmonella cleia detection kit and detection method
  • Salmonella cleia detection kit and detection method
  • Salmonella cleia detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The raw material samples used in the test were beef samples suspected to be contaminated with Salmonella.

[0035] Add 100 μl / well of Salmonella antibody at a concentration of 2 μg / mL to a 96-well ELISA plate, coat in a refrigerator at 4°C overnight, wash 3 times with phosphate washing solution PBST, pat dry, and use 200 μl of blocking solution in each well. React in a 37°C incubator for 1 hour, wash with phosphate washing solution PBST for 3 times, and then pat dry the plate for use;

[0036] Add 100 μl of the sample to be tested in each well, react in a 37°C incubator for 2 hours, wash with PBST three times, and pat dry; add horseradish peroxidase-labeled Salmonella detection antibody at a concentration of 1 μg / mL at 100 μl / well In a 96-well ELISA plate, place it in a 37°C incubator to react for 1 hour, wash it with PBST three times, and pat dry; add 100 μl / well of chemiluminescence solution, react for 5 minutes, and measure the chemiluminescence value and luminescenc...

Embodiment 2

[0038] Beef samples suspected of Salmonella contamination were detected according to the method of Example 1.

[0039] Except that iodophenol concentration is set as 0.05mM, 0.1mM, 0.15mM, 0.25mM and 0.5mM respectively; performance impact. The test results are shown in the table below:

[0040]

[0041]

[0042] The results showed that: when the molar concentration of p-iodophenol was 0.15mM, the S / N reached the highest. Therefore, when the concentration of p-iodophenol was 0.15mM, its enhancement effect on the chemiluminescence system was the best.

Embodiment 3

[0044] Beef samples suspected of Salmonella contamination were detected according to the method of Example 1.

[0045] The concentration of luminol in the system was set to 0.625mM, 1.25mM, 2.5mM, 5mM, 10mM respectively; the concentration of other components was constant, and determined by chemiluminescent enzyme immunoassay to investigate the effect of luminol concentration changes on the luminescence of chemiluminescent liquid. performance impact. The test results are shown in the table below:

[0046]

[0047] The results show that when the molar concentration of luminol is 1.25mM, the S / N reaches the highest, so the molar concentration of luminol is 1.25mM is the best luminescent performance of the chemiluminescence liquid.

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Abstract

The invention discloses a CLEIA detection kit and a detection method of salmonella. The kit includes a salmonella antibody, a horseradish peroxidase labeled salmonella antibody and a chemiluminescent solution containing p-lodophenol, luminol and hydrogen peroxide. The corresponding detection method uses the salmonella antibody coated in a 96 well microtiter plate as a capture antibody, the horseradish peroxidase labeled salmonella antibody as an antibody for detection, and luminol-H2O2 with a p-iodophenol reinforcing agent as the chemiluminescent solution, so as to establish a double antibody sandwich chemiluminescent enzyme immunoassay method for detecting content of salmonella. The method provided by the invention has the advantages of high sensitivity, good selectivity, easy operation, fast detection and low cost, can realize large-scale online detection of salmonella, and is suitable for extensive application in the fields of food safety and environmental monitoring, etc.

Description

technical field [0001] The invention relates to a detection kit and a detection method for Salmonella, more specifically, to a detection kit for detecting Salmonella by using chemiluminescent enzyme immunology technology and a detection method using the detection kit. Background technique [0002] Salmonella (Salmonella spp.) is a group of Gram-negative bacilli that parasitize in the intestinal tract of humans and animals. They have similar biochemical characteristics and antigenic structures, and are facultatively anaerobic. As a common food-borne pathogenic bacteria, Salmonella contaminates most animal-derived foods such as livestock and poultry meat, eggs, and fresh milk, and is also widely present in the natural environment. It is the main cause of food poisoning caused by microorganisms. One of the pathogens is of great significance in public health. According to reports, the Ministry of Health has issued a draft of the national standard "Limits of Pathogenic Bacteria ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569
CPCG01N33/535G01N33/56916G01N2333/255G01N2469/00
Inventor 徐静曹际娟赵亮邹明强李锦丰房芳
Owner 徐静
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