Liquid chromatography and mass spectrometry detection method for simultaneously detecting IV AIPs (auto-inducing peptides) signal molecules of staphylococcus aureus

Abstract

The invention relates to a liquid chromatography and mass spectrometry detection method for simultaneously detecting IV AIPs (auto-inducing peptides) signal molecules of staphylococcus aureus. The method includes the steps: determining liquid chromatography and mass spectrometry parameters by the aid of standard working solution of the AIPs molecules to build the liquid chromatography and mass spectrometry detection method; applying the liquid-mass chromatography detection method to detect AIPs molecules in bacterial fermentation broth of the staphylococcus aureus; detecting quorum sensing signal molecules in bacterial quorums of the staphylococcus aureus. According to the liquid chromatography and mass spectrometry method, the IV AIPs molecules are rapidly, accurately and quantitatively detected, a pretreatment method of molecule samples is simple, convenient and easy to operate, secondary pollution is avoided, detection accuracy is improved, a linear correlation coefficient and a lowest detectable limit meet detection requirements, needs of subsequent experiments can be met, and a foundation is laid for food safety monitoring.

Description

technical field [0001] The invention belongs to the technical field of bacterial microorganism detection, and in particular relates to a liquid mass spectrometry detection method for simultaneously detecting IV AIPs signal molecules of Staphylococcus aureus. Background technique [0002] With the development of science and technology, mass spectrometry is playing an increasingly important role in food safety testing. Liquid chromatography-mass spectrometry (LC-MS / MS) has incomparable advantages over traditional methods. Chromatography’s high separation ability for complex samples also applies the advantages of mass spectrometry’s high selectivity, high sensitivity, and ability to accurately provide molecular weight and structural information of compounds. In the field of food research, liquid-mass spectrometry technology can not only sensitively detect drug residues in food and harmful substances produced by additives due to chemical changes in the processing process, but al...

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Problems solved by technology

The phenomenon that bacteria communicate with each other to accomplish some important physiological functions is called quorum sensing (Qs) (Miller), and the AIPs signaling molecule is Staphylococcus aureus (S.aureus) A class of substances that transmit information in the quorum sensing system is an autoinducing peptide, which includes four structural forms according to different encodings, namely AIP-I, AIP-II, AIP-III, and AIP-IV, such as figure 1 As shown; the research of Hiyas et al. used UHPLC technology to detect the AIP-I signal molecule produced by S.aureus, and the detection limit reached 0.25μM. However, the strain used by Hiyas was not a food poisoning strain from China, and its Only studying the detection method of No. AIP-I molecule is not enough to satisfy the research on the quorum sensing system of Staphylococcus aureus, and the inventors of the present application have found that the same strain of Staphylococcus aureus can only produce one In order to study the quorum sensing of different strains, it is necessary to establish a method that can detect four kinds of AIPs molecules at the same time; in addition, the AIPs signal molecules produced by S.aureus play a role in regulating the interaction between bacteria in quorum sensing. The perception and production of physiological functions and biotoxins regulate the production of Staphylococcus aureus enterotoxin and biofilm to a certain extent, and enterotoxin and biofilm are the main causes of hidden dangers to food microbial safety. A rapid and timely method for simultaneous detection of IV AIPs molecules is necessary to achieve early detection of toxin and biofilm production

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