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Construction method for MALDI-TOF-MS database of common pathogenic bacteria in food and animal products

A MALDI-TOF-MS, animal product technology, applied in the field of MALDI-TOF-MS database construction, can solve problems such as the mass spectrum database of bacterial strains that have not yet been found, and achieve the effect of accurate identification

Inactive Publication Date: 2012-06-27
INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since the research and application of this technology in China is still in its infancy, there have been no reports on the establishment of mass spectrogram databases for various bacterial strains.

Method used

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  • Construction method for MALDI-TOF-MS database of common pathogenic bacteria in food and animal products
  • Construction method for MALDI-TOF-MS database of common pathogenic bacteria in food and animal products
  • Construction method for MALDI-TOF-MS database of common pathogenic bacteria in food and animal products

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Embodiment 1

[0016] A method for constructing a MALDI-TOF-MS database of common pathogenic bacteria in food and animal products in this embodiment includes the following steps: 1) sample processing; 2) data collection; 3) data analysis; 4) bacterial database of establishment.

[0017] The treatment method of the sample in the step 1) is: use the liquid medium to enrich the bacteria and then do the MALDI-TOF-MS test; the liquid medium, when the bacteria used are Vibrio, inoculate Other bacteria were inoculated in a liquid medium of nutrient broth in a liquid medium of alkaline protein frozen water. All were purchased from Beijing Land Bridge Technology Co., Ltd.

[0018] The specific steps of the MALDI-TOF-MS test are: take 1 mL of the culture solution in a 1.5 mL centrifuge tube, centrifuge at 10,000 rpm for 2 min, pour off the supernatant, add 1 mL of sterilized normal saline, mix well, and centrifuge at 10,000 rpm for 2 min. min, discard the supernatant, and wash again to fully remove ...

Embodiment 2

[0024] Identification of Salmonella in food and animal products

[0025] The MALDI-TOF MS method of the present embodiment detects and identifies the process of Salmonella, comprises the following steps:

[0026] 1. Enrichment culture and purification

[0027] According to the corresponding methods of enrichment and purification of Salmonella at home and abroad, the Salmonella in food and animal samples are enriched and purified, and a single suspicious colony is taken and inoculated in the nutrient broth, cultivated for 18-24 hours before use.

[0028] 2. Treatment of colony samples Take 1 mL of nutrient broth culture solution (purchased from Beijing Land Bridge Technology Co., Ltd., item number: CM106, batch number: 110212) in a 1.5 mL centrifuge tube, centrifuge at 10,000 rpm for 2 min, discard the supernatant, and add 1 mL of sterilized normal saline, mix thoroughly, centrifuge at 10,000 rpm for 2 min, discard the supernatant, and wash again to fully remove the medi...

Embodiment 3

[0034] Identification of Vibrio parahaemolyticus in aquatic products

[0035] The process of detecting and identifying Vibrio parahaemolyticus by the MALDI-TOF MS method of the present embodiment comprises the following steps:

[0036] 1. Enrichment culture and purification

[0037] According to the corresponding enrichment and purification methods of Vibrio parahaemolyticus at home and abroad, culture and purify Vibrio parahaemolyticus in aquatic products, take a single suspicious colony and inoculate it in alkaline peptone water with 3.5% salt, (Purchased from Beijing Land Bridge Technology Co., Ltd., item number: CM401, batch number: 100524) After culturing for 18-24 h, it is ready for use.

[0038] 2. Processing of Colony Samples

[0039] Take 1 mL of culture solution in a 1.5 mL centrifuge tube, centrifuge at 10,000 rpm for 2 min, discard the supernatant, add 1 mL of sterilized normal saline, mix well, centrifuge at 10,000 rpm for 2 min, discard the supernatant,...

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Abstract

The invention discloses a construction method for MALDI-TOF-MS database of common pathogenic bacteria in food and animal products. The method comprises the following steps: 1) processing of a sample; 2) acquisition of data; 3) data analysis; and 4) establishment of bacteria database. According to the invention, the MALDI-TOF-MS database of common pathogenic bacteria, including vibrio cholerae, which are pathogenic to human beings and animals in domestic food and animal products is established and can be used for rapid detection and identification of a variety of human and animal pathogens.

Description

technical field [0001] The invention relates to a method for constructing a MALDI-TOF-MS database of common pathogenic bacteria in food and animal products. Background technique [0002] Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) technology based on molecular weight detection of bacterial surface proteins is a new rapid detection and identification technology for microorganisms. By measuring the unique protein composition of bacteria, mass spectrometry is used to arrange the measured proteins and peptides in sequence according to molecular weight, forming a unique proteome fingerprint, and identifying strains through characteristic model peaks. The basic principle is: point the microorganisms and the same amount of matrix on the sample plate, the solvent volatilizes to form a co-crystal of the sample and the matrix, the matrix absorbs energy from the laser to desorb the sample, and the charge transfer between the matrix and th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/64G06F19/18
Inventor 龚艳清郭书林陈信忠杨俊萍叶妍妍孔繁德徐淑菲
Owner INSPECTION & QUARANTINE TECH CENT OF XIAMEN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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