Test paper for salmonella and preparation method of test paper

A Salmonella, detection test paper technology, applied in the field of detection, can solve the problems of time-consuming, labor-intensive sensitivity and specificity, complicated procedures, high missed detection rate, etc., and achieve the effect of good stability, simple operation, and no need for equipment

Inactive Publication Date: 2016-10-12
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The detection of Salmonella has always been based on traditional detection methods. Although conventional methods such as non-selective and selective enrichment and isolation of suspicious bacteria are classic and reliable, the procedures are complex and cumbersome, not only time-consuming, but also poor in sensitivity and specificity, and missed detection. higher rate
Immunofluorescence, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR) and other methods require the use of designated equipment, corresponding test conditions and skills, and are difficult to promote at the grassroots level

Method used

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  • Test paper for salmonella and preparation method of test paper
  • Test paper for salmonella and preparation method of test paper
  • Test paper for salmonella and preparation method of test paper

Examples

Experimental program
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Embodiment 1

[0063] Example 1 Preparation and Purification of Antibodies

[0064] Salmonella was activated with a special medium, and characteristic colonies appeared after culture. Two strains of Salmonella were expanded and cultured, and preserved for later use. The measured bacterial concentration was about 1×10 10 CFU, bacteria are inactivated with 0.3% formaldehyde for 24 hours, after the inactivation, the bacteria liquid will no longer grow bacteria on the bacteria test plate. The inactivated bacterial solution was added with Freund's incomplete adjuvant to make a vaccine for immunizing New Zealand white rabbits, 1mL / only, subcutaneously injected on the back, immunized 4 times in total, with an interval of 15 days between each time, and injected 4 times in a row using the same method. Before the fourth immunization, blood was collected from the rabbit's ear vein to detect the antibody production. Blood was collected from the carotid artery of those who passed the test, and the ser...

Embodiment 2

[0065] The preparation of embodiment 2 colloidal gold solution

[0066] Take 100ml of double-distilled water and heat to boil, add 1ml of 1% chloroauric acid solution, shake well, continue to heat and boil for 3min, then quickly add 1ml of 1% trisodium citrate solution, transfer to a magnetic stirrer and stir for 2min, at this time The color changes from gray to black, and then to purple. Put it on the electric stove and boil for 6 minutes, and the color turns into wine red. After cooling, distilled water is added to make the volume to 100ml. The colloidal gold solution should be protected from light and stored at 4°C.

Embodiment 3

[0067] Preparation and purification of colloidal gold-labeled Salmonella of embodiment 3

[0068] Get the colloidal gold solution that needs to prepare amount, use the K of 0.2mol / L 2 CO 3 Or 0.1mol / L HCl solution to adjust the pH of the colloidal gold solution to 8.0. Then add 9.6ug of purified rabbit antibody per 1mL of colloidal gold solution, magnetically stir and mix evenly, add 10% PEG20000 to a final concentration of 1%, after mixing, let it stand overnight at 4°C to obtain the labeled gold-labeled antibody. Centrifuge the gold-labeled monoclonal antibody at 2000r / min, 4°C for 20min, discard the precipitate; centrifuge at 10000r / min, 4°C for 30min, discard the supernatant. With 0.05mol / L TBS (containing 1% BSA, 0.05% NaN 3 ) buffer solution to dissolve the precipitate to the original volume, repeat centrifugation 2-3 times, dissolve the precipitate in 1 / 10 of the original volume of TBS, and store at 4°C for later use.

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Abstract

The invention relates to test paper for salmonella and a preparation method of the test paper. The test paper for the salmonella comprises a bottom plate, the bottom plate is provided with a first end and a second end and provided with a sample pad, a colloidal gold pad, a nitrocellulose membrane and a water absorption pad in sequence from the first end to the second end, the colloidal gold pad is provided with salmonella antibodies marked with colloidal gold, a detection line and a comparison line are further formed on the nitrocellulose membrane, the detection line is prepared from a salmonella antibody membrane capable of being combined with the salmonella, and the comparison line is prepared from a goat anti-rabbit antibody membrane capable of being combined with the salmonella antibodies.

Description

technical field [0001] The invention relates to a detection method, in particular to a detection test paper for Menella and a preparation method thereof. Background technique [0002] Salmonella is a common zoonotic intestinal pathogen, which can not only cause diseases such as typhoid and paratyphoid in livestock and poultry, but also cause common diseases such as diarrhea and bacterial food poisoning in humans, threatening human health. There are more than 2,000 known serotypes of Salmonella, which are widely distributed in nature. The serotypes related to human diseases are mainly concentrated in groups A to E, among which Salmonella typhimurium, Salmonella choleraesuis and Salmonella enteritidis are the most common. Among all kinds of bacterial food poisoning in various countries in the world, food poisoning caused by Salmonella ranks at the forefront. Human and animal infection with Salmonella may aggravate the disease or increase mortality, and may also reduce animal...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531G01N33/532
CPCG01N33/56916G01N33/531G01N33/532G01N2333/255
Inventor 李俊彭喆黄保华时建立王莉莉王金宝徐绍建吴晓燕朱晓琳郑书轩张玲玲
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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