Molecular motor biosensor kit for molecular subtyping of salmonella

A biosensor and molecular motor technology, applied in the field of molecular motor biosensor kits for Salmonella molecular typing, can solve problems such as complicated procedures and cumbersome

Inactive Publication Date: 2013-02-06
BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT +1
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  • Abstract
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Problems solved by technology

However, the above-mentioned methods are relatively cumberso

Method used

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  • Molecular motor biosensor kit for molecular subtyping of salmonella
  • Molecular motor biosensor kit for molecular subtyping of salmonella
  • Molecular motor biosensor kit for molecular subtyping of salmonella

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Embodiment

[0042] (1) Bacterial culture and DNA extraction

[0043] 12 strains of Salmonella were isolated from food. 12 strains of Salmonella were inoculated into buffered peptone-containing water (BPW) for enrichment, and after culturing at 36°C ± 1°C for 8-18 hours, 1 mL of the enrichment solution was inoculated into 10 mL of sodium tetrathiosulfonate brilliant green (TTB) enrichment solution and In selenite cystine (SC) enrichment solution, incubate at 42℃±1℃ and 36℃±1℃ for 18-24h, then streak inoculated with bismuth sulfite (BS) and xylose lysine deoxygenation Bile salt (XLD) agar. Finally, the suspicious colonies were picked and transferred to nutrient agar, and cultured at 36℃±1℃ for 24h for DNA extraction.

[0044] The cultured Salmonella was extracted with a DNA extraction kit (spin column type) DP320, and the detailed operation steps were shown in the kit instructions. The liquid in the centrifuge tube finally obtained is the bacterial DNA extract, which is used as a templat...

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Abstract

The invention belongs to a kit for quickly performing molecular subtyping determining to salmonella, and particularly relates to the technology of utilizing an F0F1-ATPase molecular motor biosensor Chro-invA, Chro-hut, Chro-spvR and Chro-sdf in the kit to detect specific gene invA and hut of the salmonella, toxic plasmid gene spvR and salmonella enteritidis strain specific gene sdf, so as to determine whether the salmonella contains toxic plasmid and whether the salmonella is the salmonella enteritidis in a subtyping way. The kit has functions that 1, the invA and the hut of the salmonella is detected, so as to identify target bacterium; 2, the spvR can be detected to determine whether the target bacterium carries the toxic plasmid; and 3, the sdf is detected to determine whether the target bacterium is the salmonella enteritidis. The method can be used for establishing the F0F1-ATPase molecular motor biosensor in the kit, carrying out molecular subtyping of the F0F1-ATPase molecular motor biosensor, and determining whether the salmonella contains the toxic plasmid and whether the salmonella is the salmonella enteritidis in a subtyping way according to a subtyping result. The method has the advantages of being high in sensitivity, fast, simple to operate, and low in cost.

Description

technical field [0001] The invention belongs to a kit for rapid molecular typing judgment of Salmonella, specifically using F in the kit 0 F 1 -ATPase molecular motor biosensors Chro-invA, Chro-hut, Chro-spvR, Chro-sdf detect Salmonella genus-specific genes invA, hut, virulence plasmid gene spvR and Salmonella Enteritidis species-specific gene sdf, In order to judge whether Salmonella carries a virulent plasmid and whether it is Salmonella Enteritidis. Background technique [0002] Salmonella is a kind of pathogenic bacteria ubiquitous in nature. In food poisoning all over the world, the poisoning cases caused by Salmonella occupy the first or second place. The diseases it causes are mainly divided into two categories: one is severe infectious diseases such as typhoid fever, and the other is acute gastroenteritis. Common meat foods and dairy and egg foods can be contaminated by Salmonella. Among them, Salmonella typhimurium, Salmonella choleraesuis, and Salmonella enterit...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/10G01N21/64
CPCY02A50/30
Inventor 张捷周琦王静卢晓宇李兆杰张惠媛齐玮刘岩汪琦张昕陈广全王小晋王佩荣乐加昌
Owner BEIJING ENTRY EXIT INSPECTION & QUARANTINE BUREAU INSPECTION & QUARANTINE TECH CENT
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