Method for evaluating vibrio parahaemolyticus VBNC state in digestive tract in vitro

By simulating the gastrointestinal fluid environment and combining multiple detection methods to assess the growth, metabolism, and VBNC status of Vibrio parahaemolyticus, the problem of insufficient assessment in existing technologies has been solved, and a comprehensive assessment and risk prediction of Vibrio parahaemolyticus in the digestive tract has been achieved.

CN120829949APending Publication Date: 2025-10-24BEIJING TECH & BUSINESS UNIV +1
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Patent Information

Application Number
CN202510782701.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-06-12
Publication Date
2025-10-24

AI Technical Summary

Technical Problem

Existing technologies cannot accurately reflect the growth and metabolism of Vibrio parahaemolyticus in the gastrointestinal tract and whether it has entered the VBNC state, leading to missed and false detections and increasing public health risks.

Method used

Using a simulated gastrointestinal fluid environment, plate counting, PMAxx-PCR, flow cytometry, and ATP detection were employed to assess the growth and metabolism of Vibrio parahaemolyticus under different pH values ​​and nutrient systems. Cell membrane permeability was observed using SEM to systematically evaluate its VBNC status.

Benefits of technology

This study enables a comprehensive assessment of the growth and metabolism of Vibrio parahaemolyticus in the digestive tract, accurately determines its VBNC status and potential resuscitation risks, provides a basis for prevention and control strategies, and reduces the risk of infection.

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Abstract

The method for evaluating the vibrio parahaemolyticus VBNC state in the digestive tract in vitro comprises the following steps that simulated gastric juice and vibrio parahaemolyticus bacterial liquid are added into a centrifugal tube, test groups 1, 2 and 3 without glucose are set to be 2.5, 3.5 and 4.5 in pH value and test groups 4, 5 and 6 with the pH value being 2.5, 3.5 and 4.5 and 4 g of glucose are set to be 2.5, 3.5 and 4.5 respectively, after constant-temperature shaking bath culture is carried out for 120 min, simulated intestinal juice is added, then culture continues to be carried out for 120 min, and the vibrio parahaemolyticus VBNC state in the digestive tract is obtained. The simulated digestive juice is extracted at 30 min, 60 min, 120 min and 240 min, a plate counting method, PMAxx-qPCR, flow cytometry, ATP detection, SEM observation and cell membrane permeability detection are conducted on the simulated digestive juice, and the growth and metabolism conditions of the vibrio parahaemolyticus in the digestive tract and the condition that the vibrio parahaemolyticus enters the VBNC state are evaluated; therefore, rapid and systematic evaluation of the vibrio parahaemolyticus in the digestive tract is realized.
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