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Salmonella detection method and application thereof

A Salmonella detection method technology, applied in the detection field of Salmonella, can solve the problems of cumbersome operation, long detection time, and large error in result judgment, and achieve the effect of low cost, high sensitivity, and short detection time

Inactive Publication Date: 2018-09-14
江西华牧生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, many countries, including my country, still use the traditional culture method for the clinical detection of Salmonella. This conventional Salmonella detection has many disadvantages such as cumbersome operation, long detection time, and large error in result judgment, which is far from being able to meet the modern rapid The need for testing and diagnosis
In recent years, Salmonella detection technology has made great progress, from the very difficult traditional isolation and culture method to immunological detection method to today's molecular biology detection technology, such as the loop-mediated isothermal amplification technology studied by Huang Jinhai et al. (LAMP) to detect Salmonella ("Establishment of a rapid detection method for Salmonella LAMP in food", "Journal of Tianjin University", 2012, issue 05), Zhong Weijun et al. studied the establishment of a rapid detection method for Salmonella in food ("Food Salmonella PCR rapid detection method", "Chinese Journal of Zoonoses", No. 12, 2007), however, the above molecular detection methods require expensive equipment, and are not intuitive in the judgment of the results, and need certain equipment to judge

Method used

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  • Salmonella detection method and application thereof
  • Salmonella detection method and application thereof
  • Salmonella detection method and application thereof

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Embodiment 1

[0017] Embodiment 1: a kind of detection method of Salmonella comprises the steps:

[0018] Step 1, preparing probe molecules: according to the genome structure and conserved sequence of Salmonella, select the invA gene of Salmonella as the detection target fragment, according to the Salmonella invA gene sequence disclosed in GenBank: EU348365, design probe molecules, the probe molecule sequence is 5'-GCGAGTGGTAAATTATTCCGATGAAGT-3', the 5' end of the probe molecule is labeled with the reporter fluorescent group FAM, that is, the molecular structure of the probe is: FAM-GCGAGTGGTAAATTATTCCGATGAAGT;

[0019] In step two, the polystyrene nanosphere solution is mixed with the probe molecule prepared in step one: FAM-GCGAGTGGTAAATTATTCCGATGAAGT, and then added to the sample to be tested to detect fluorescence.

Embodiment 2

[0020] Embodiment 2: a kind of detection method of Salmonella comprises the steps:

[0021] Step 1, preparing probe molecules: according to the genome structure and conserved sequence of Salmonella, select the invA gene of Salmonella as the detection target fragment, according to the Salmonella invA gene sequence disclosed in GenBank: EU348365, design probe molecules, the probe molecule sequence is 5'-GCGAGTGGTAAATTATTCCGATGAAGT-3', the 5' end of the probe molecule is labeled with the reporter fluorescent group FITC, that is, the molecular structure of the probe is: FITC-GCGAGTGGTAAATTATTCCGATGAAGT;

[0022] In step two, the polystyrene nanosphere solution is mixed with the probe molecule prepared in step one: FITC-GCGAGTGGTAAATTATTCCGATGAAGT, and then added to the sample to be tested to detect fluorescence.

Embodiment 3

[0023] Embodiment 3: A kind of test kit for Salmonella detection, it comprises: probe molecule solution, polystyrene nanosphere solution, positive control plasmid, negative control, wherein the structure of said probe molecule is: reporter fluorescent group -GCGAGTGGTAAATTATTCCGATGAAGT.

[0024] Wherein the positive control plasmid is the method of PCR amplification, the sequence is GenBank: GenBank: EU348365 disclosed Salmonella invA gene cloned on the pGEM-T Easy vector, the positive plasmid obtained after verification; wherein the negative control group is water.

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Abstract

The invention relates to a salmonella detection method. The method includes the steps of: 1, preparing a probe molecule: according to the genome structure and conserved sequence of salmonella, selecting the invA gene of salmonella as the detection target fragment, according to the salmonella invA gene sequence disclosed by GenBank:EU348365, designing a probe molecule, which has a sequence of 5'-GCGAGTGGTAAATTATCGATGAAGT-3', with the 5' end being labeled with a reporter fluorophore; and 2, mixing a polystyrene nanosphere solution with the probe molecule prepared by step 1, then adding a to-be-detected sample, and performing fluorescence detection.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to the detection of Salmonella, a pathogenic bacterium of food-borne diseases, in particular to a method for detecting Salmonella and an application thereof. Background technique [0002] Salmonella (Salmonella) is a pathogen discovered more than a hundred years ago. It is a large genus of Enterobacteriaceae with similar morphology and cultural characteristics. It has various types and is widely distributed. It is a common and important zoonosis It can not only cause gastroenteritis, but also various syndromes such as typhoid fever, sepsis, and extraintestinal focal infection. According to statistics, food poisoning caused by Salmonella often ranks first in bacterial food poisoning in countries around the world. In my country, food poisoning caused by Salmonella also ranks first in bacterial food poisoning. Salmonella is an important indicator for monitoring food hygiene status. Once S...

Claims

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Application Information

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IPC IPC(8): C12Q1/6816C12Q1/10
CPCC12Q1/6816C12Q2563/107C12Q2563/149C12Q2563/155Y02A50/30
Inventor 陈瑞张志刚张磊
Owner 江西华牧生物科技有限公司
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