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Method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in food simultaneously and rapidly

A technology for Salmonella typhi and paratyphoid B, which is applied in the field of microbial detection, can solve the problems of high labor intensity, time-consuming and high cost, and achieves the effects of simple operation, high efficiency and simple result judgment.

Inactive Publication Date: 2012-09-19
NANCHANG UNIV
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Problems solved by technology

For example (Yang Ruijun, Analysis of the detection results of pathogenic bacteria in raw meat food [J] China Health Inspection Journal. 2009:19 (9) 2122-2125), the traditional method has the disadvantages of high labor intensity, time-consuming, and high cost

Method used

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  • Method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in food simultaneously and rapidly
  • Method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in food simultaneously and rapidly
  • Method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in food simultaneously and rapidly

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Embodiment

[0031] Step 1, Food Sample Pretreatment

[0032] Weigh 1 g of food sample, add 9ml of PBS, grind to make a homogenate, centrifuge at 900 g for 5 min, remove the enlarged food residue, and take the supernatant (this process must be performed aseptically).

[0033] Step 2, PMA processing

[0034] PMA was dissolved in dimethyl sulfoxide (DMSO), prepared into a 0.5 mg / mL PMA solution, and stored at -20°C in the dark. Take 500 μL of the prepared bacterial suspension and place it in a 1.5 mL microcentrifuge tube, add 3 μL of 0.5 mg / mL PMA solution to make the final mass concentration of PMA 3 μg / mL; Incubate in the dark at room temperature for 5 min, and expose to a 500 W halogen lamp for 5 min. When cross-linking with light, place the sample on ice (to avoid overheating), and place the cross-linked suspension at 10,000 g at a distance of 20 cm from the light source. After centrifugation for 5 min, the resulting pellet was used for DNA extraction.

[0035] Step 3, Genomic DNA Ext...

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Abstract

The invention belongs to the field of microbiological detection and discloses a method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in food simultaneously and rapidly. A special primer is designed aiming at salmonella typhimurium, salmonella paratyphi b and salmonella typhi, hydrazoic brominated c ingot processing is adopted to remove interference of dead bacteria, and finally deoxyribonucleic acids (DNAs) are extracted to perform multiplex polymerase chain reaction. Compared with a classical bacterium separating and identifying method and a serologic typing method, the method for detecting live salmonella typhimurium, salmonella paratyphi b and salmonella typhi in the food simultaneously and rapidly is rapid and accurate.

Description

technical field [0001] The invention belongs to the field of microorganism detection, in particular to a method for rapidly detecting live Salmonella typhimurium, Salmonella paratyphi B and Salmonella typhi simultaneously in food. technical background [0002] Salmonella species are Gram-negative, facultatively anaerobic, non-spore-forming bacteria. More current classification, Salmonella only includes two species: Salmonella Bongor and Salmonella enterica, the latter contains more than 2,500 serotypes, but also the most infecting humans and animals. Among the many serotypes, Salmonella typhimurium is an important pathogen that causes foodborne pathogenic bacteria poisoning and gastroenteritis in humans. It is mainly prevalent in developing countries, and for immunocompromised individuals, if Infection with this pathogen can be fatal if left untreated. Salmonella typhi and paratyphi mainly cause typhoid fever and paratyphoid fever in humans. Although they are no longer p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12R1/42
Inventor 魏华许恒毅万翠香杨友均徐锋熊勇华赖卫华
Owner NANCHANG UNIV
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