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A method for quantitative determination of Vibrio parahaemolyticus in aquatic products using internal standard proteome

A technology of proteome and Vibrio hemolyticus, which can be used in measurement devices, instruments, scientific instruments, etc., and can solve problems such as quantification

Active Publication Date: 2020-02-25
INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, some physical and chemical properties, pathogenicity and stress resistance of Vibrio parahaemolyticus are improved to varying degrees (for example, at 4°C, its anti-peroxide, hydrogen peroxide, anti-ultraviolet, anti-chemical reagents, etc.) , and low temperature adaptability have been improved, mainly relying on the quantitative analysis of the specific changes in the proteome of the bacteria in the low temperature environment, after the discovery of the above conclusions) before the harm of Vibrio parahaemolyticus, only some conventional colony cultures were used Evaluation with biochemical means, there are many demonstrations in the way of identification, but it has not been able to fundamentally carry out scientific quantification

Method used

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  • A method for quantitative determination of Vibrio parahaemolyticus in aquatic products using internal standard proteome
  • A method for quantitative determination of Vibrio parahaemolyticus in aquatic products using internal standard proteome
  • A method for quantitative determination of Vibrio parahaemolyticus in aquatic products using internal standard proteome

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The method for quantitative determination of Vibrio parahaemolyticus in aquatic products with the internal standard method proteome quantitative determination provided in this embodiment comprises the following steps:

[0040] (1) Identification of strains and culture medium

[0041] The isolated Vibrio parahaemolyticus should be identified by the GN card system of VITEK 2 Compact or other microbial identification instruments, and the reliability should reach more than 95%;

[0042] (2) Isotope labeling and cold shock process

[0043] After the pure culture of the strain was diluted with normal saline, it was streaked and inoculated on a tryptone yeast extract soybean agar plate (TSA-YE, Difco), and incubated at 37°C for 24 hours; then two colonies were taken and inoculated separately 14 N and 15 N medium, inoculated 14 The medium of N was cultured at 4°C for 2 weeks as the experimental test strain (the reason for this is that this condition is closer to the temperat...

Embodiment 2

[0064] Embodiment 2 result is as follows:

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073] From the above experimental results, the following conclusions can be drawn:

[0074] 1. The increase of peroxide kinase indicates that the bacteria's ability to resist peroxides, such as hydrogen peroxide, is improved. Hydrogen peroxide is one of the most commonly used disinfectants in aquatic product processing. If the conventional 2%-5% volume concentration of hydrogen peroxide is used to When it is used in the bacterial control method in the production and processing of aquatic products, the effect will definitely be greatly affected. It will be mistaken for the illusion that Vibrio parahaemolyticus died after soaking in conventional hydrogen peroxide.

[0075] 2. The aromatic amino acid synthesis pathway is one of the important ways for microorganisms to interact with the external environment. It is also closely related to the...

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Abstract

The invention discloses a method for quantifying and determining vibrio parahaemolyticus in an aquatic product by utilizing an internal-standard-method proteome. The method comprises the following steps: (1) identifying a strain and preparing a culture medium; (2) carrying out an isotope labeling and cold shocking process; (3) preparing a sample. According to the method disclosed by the invention, the vibrio parahaemolyticus in the aquatic product is quantified and determined by utilizing the internal-standard-method proteome; compared with other non-labeling or chemical methods, an isotope internal standard has the greatest advantage that a series of errors caused by unavoidable negative effects including reagents, operation, purity and the like in an experiment process are reduced to the greatest extent; especially, in a protein operation process, N15 and N14 are directly mixed to form a sample for analyzing and a whole system is formed, so that errors caused by dual-system analysis are reduced to the greatest extent.

Description

technical field [0001] The invention relates to a method for quantifying Vibrio parahaemolyticus in aquatic products by using an internal standard method proteome. Background technique [0002] Vibrio parahaemolyticus is a food-borne pathogen that is highly susceptible to infection in various links of aquatic product processing, transportation, and storage. Its pathogenicity can cause acute intestinal diseases in humans and spread on a large scale, causing great harm. Especially in raw or semi-raw seafood, such as oysters, sashimi, etc., the risk will be higher. Moreover, some physical and chemical properties, pathogenicity and stress resistance of Vibrio parahaemolyticus are improved to varying degrees (for example, at 4°C, its anti-peroxide, hydrogen peroxide, anti-ultraviolet, anti-chemical reagents, etc.) , and low temperature adaptability have been improved, mainly relying on the quantitative analysis of the specific changes in the proteome of the bacteria in the low t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 马维兴黄小华贾俊涛吴振兴唐静赵丽青孙宇马云
Owner INSPECTION & QUARANTINE TECH CENT SHANDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU