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A kind of detection method, test strip and application of Salmonella enteritidis

A Salmonella enteritidis and detection method technology, applied in the field of biological detection, can solve the problems that affect the wide application of immunochromatographic test strips, complex nanomaterial synthesis methods, and affect antibody activity, and achieve simple structure, low cost, and novel methods Effect

Active Publication Date: 2022-03-11
NORTHWEST A & F UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the sensitivity is improved, the synthesis methods of these nanomaterials are more complicated and the synthesis conditions are higher
In addition, during the construction of traditional probes, the activity of antibodies will be affected, which in turn will affect the wide application of immunochromatographic test strips

Method used

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  • A kind of detection method, test strip and application of Salmonella enteritidis
  • A kind of detection method, test strip and application of Salmonella enteritidis
  • A kind of detection method, test strip and application of Salmonella enteritidis

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1: Preparation method of Salmonella enteritidis universal monoclonal antibody

[0039] The preparation method of Salmonella enteritidis universal monoclonal antibody comprises the following steps:

[0040] 1) Animal immunity

[0041] Purchase 6-week-old BALB / c mice, and use the Salmonella enteritidis preserved in our laboratory provided by Yang Baowei's laboratory to extract flagellin for immunization. For the first immunization, 0.8 mg / mL Salmonella enteritidis flagellin was mixed and emulsified with an equal amount of Freund's complete adjuvant, and the mice were injected intraperitoneally with the emulsified antigen. The second immunization was carried out 4 weeks later, using 0.8 mg / mL Salmonella enteritidis flagellin mixed with an equal amount of Freund's incomplete adjuvant to emulsify, and the mice were intraperitoneally injected with the emulsified antigen. The third immunization was carried out 4 weeks later, and the immunization method was the sam...

Embodiment 2

[0057] Example 2: Preparation method of nitrogen-rich carbon nanoparticles with positive surface charge

[0058] The preparation method of the nitrogen-rich carbon nanoparticles with positive surface charge, comprising the following steps:

[0059] 1) Preparation of nitrogen-rich carbon nanoparticles

[0060] The urea was placed in a covered crucible, heated in air for 4 hours to 550°C and held at this temperature for a further 4 hours. The yellow solid formed was carbon nitride (g-C3N4), which was then ground into powder for further use. Prepare a homogeneous precursor solution by dissolving 1,4-phthalic acid and diamine triacetate in N,N-dimethylformamide (DMF). Carbon nitride powder is then added to the mixed precursor solution under magnetic stirring. After removal of the solvent, the solid powder was transferred to a tube furnace, heated slowly to 800 °C under argon, and maintained at this temperature for 1 h. The obtained black solid powder is nitrogen-enriched carbo...

Embodiment 3

[0064] Embodiment 3: each condition optimization of the test strip of rapid detection Salmonella enteritidis

[0065] 1) Preparation of nitrocellulose membrane (combined with figure 1 and figure 2 )

[0066] Coating of detection line: Dissolve Salmonella enteritidis monoclonal antibody in the coating solution to prepare a 1mg / mL solution; use the streaking method to coat the coating solution laterally at the edge of the nitrocellulose membrane at a speed of 1μL / cm 30mm position (that is, the detection line), and then dried at 37°C for 30 minutes. The coating liquid is: 0.02g sodium azide, 0.8g sodium chloride, 0.29g disodium hydrogen phosphate dodecahydrate, 0.02g potassium chloride, 0.02g potassium dihydrogen phosphate and dilute to 100mL with water.

[0067] 2) Preparation of the sample pad: cut the glass fiber membrane to a size of 15mm in length and 3mm in width, soak it in the blocking solution, and dry it at 37°C for 10-16 hours to obtain a sample pad, and then store...

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Abstract

The invention discloses a method for detecting Salmonella enteritidis, a test strip and its application. The method for detecting Salmonella enteritidis comprises incubating positively charged nitrogen-rich carbon nanoparticles with Salmonella enteritidis in a sample to be detected to obtain a solution to be tested, and then The Salmonella enteritidis bound to the positively charged nitrogen-rich carbon nanoparticles in the test solution is captured by the Salmonella enteritidis monoclonal antibody. The invention only uses one antibody to draw on the nitrocellulose membrane for direct detection, which breaks the traditional sandwich detection method that uses two antibodies at the same time, greatly saves the cost, solves the difficulty of pairing antibodies, and is simpler, more convenient and novel.

Description

technical field [0001] The invention belongs to the field of biological detection, and specifically relates to a detection method for Salmonella enteritidis, a test strip and an application thereof. Background technique [0002] Salmonella is a kind of Gram-negative, most of the perinatal flagella, can move, no capsule and spores, short bacilli with blunt ends, the optimum growth temperature is 37 ℃, and can reproduce normally above 20 ℃. Salmonella infection is one of the zoonotic diseases of great significance in public health. Food transmission in summer and autumn is the main route of infection. Diarrhea, fever, headache, abdominal pain, nausea and vomiting generally occur within 12-14 hours after human ingestion Symptoms such as chills and chills are accompanied by symptoms such as fatigue and muscle aches, which can cause death in severe cases. Among them, Salmonella typhimurium, Salmonella choleraesuis and Salmonella enteritidis are the most common. Salmonella is a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/577G01N33/558
CPCG01N33/54346G01N33/558G01N33/56916G01N33/577G01N2333/255
Inventor 王建龙王宗汉张道宏姚晓琳窦磊娜赵兵欣
Owner NORTHWEST A & F UNIV
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