Salmonella specific forward and reverse primers and probe, detection kit and application of detection kit

A detection kit, forward and reverse primer technology, applied in the detection field, can solve the problems of high requirements on the operating level of the staff and the inability to meet the requirements of on-site testing, and achieve the effects of instant on-site testing, rapid on-site testing, and high accuracy.

Inactive Publication Date: 2020-04-10
JIANGSU OCEAN UNIV +1
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Problems solved by technology

However, this method requires expensive instruments and equipment, requires a high level of operation of the staff, and cannot meet the on-site detection requirements of grassroots medical organizations and remote areas. For this reason, the present invention uses the RPA-LFS technology to quickly identify Salmonella, in order to provide The detection of this bacteria brings new technical reference

Method used

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  • Salmonella specific forward and reverse primers and probe, detection kit and application of detection kit
  • Salmonella specific forward and reverse primers and probe, detection kit and application of detection kit
  • Salmonella specific forward and reverse primers and probe, detection kit and application of detection kit

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Embodiment

[0031] experiment material

[0032] The genomes of standard strains of Salmonella, Staphylococcus aureus, Enteropathogenic Escherichia, Listeria monocytogenes, and Bacillus cereus were all provided by Wuhan Food and Cosmetic Quarantine Institute; Basic kit, nfokit was purchased from TwistDX Company in the UK; lateral flow chromatography test strips were purchased from Hangzhou Ustar Biotechnology Co., Ltd.; PCR cleaning kit was purchased from Mona Biotechnology Co., Ltd.; Qubit 4 was purchased from Thermo Scientific; MonAmp TM SYBR Green was purchased from Mona Biotechnology Co., Ltd.; primers and probes used in this study were synthesized by General Biosystems (Anhui) Co., Ltd.; Roche LightCycler480 fluorescent quantitative PCR instrument was purchased from Roche Pharmaceuticals.

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Abstract

The invention relates to the technical field of detection, in particular, to salmonella specific forward and reverse primers and probe, a detection kit and an application of the detection kit. The detection kit includes the forward and reverse primers, the probe and a lateral flow chromatography test strip; the 5' end of the reverse primer is labeled by using biotin; and the 5' end of the probe islabeled by using fluorescein isothiocyanate (FITC), a C base in the middle region (at least 30 bp away from the 5' end and at least 15 bp away from the 3' end) is replaced by using [THF[, and the 3'end is end-capped by using C3-spacer. The detection kit can realize real-time on-site immediate and quick field detection by using the isothermal recombinase polymerase amplification and the lateral flow strip (RPA-LFS) method in the non-laboratory environment without relying on instrument equipment.

Description

technical field [0001] The invention relates to the technical field of detection, in particular to a Salmonella-specific forward and reverse primer and probe, a detection kit and applications thereof. Background technique [0002] Salmonella is a Gram-negative pathogen widely distributed in nature. According to statistics, food poisoning caused by Salmonella often ranks first among bacterial food poisoning in various countries in the world. Salmonella is the first in my country's inland areas. The transmission route of Salmonella is mainly food intake, such as contaminated meat, eggs and egg products, dairy products, seafood and so on. Salmonella is extremely harmful to the health of humans and animals. Humans will have symptoms such as nausea, vomiting, diarrhea, abdominal pain or cramps, fever, and headache 12-36 hours after eating contaminated food. Therefore, accurate, fast and simple identification of Salmonella can not only improve the quality of food, avoid Salmonel...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/10C12Q1/06C12R1/42
CPCC12Q1/689C12Q1/6844C12Q2521/507C12Q2522/101C12Q2565/625
Inventor 许恒皓郦娟董井泉吴华华王锦鑫赵盼盼吉敬
Owner JIANGSU OCEAN UNIV
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