Detecting method of vibrio parahemolyticus in coastal seawater
A hemolytic vibrio and detection method technology, which is applied in the field of detection of vibrio parahaemolyticus in coastal seawater, can solve the problems of long time consumption, high manpower and material resources consumption, and inability to quantify seawater, so as to shorten the detection time and improve the accuracy Effect
Inactive Publication Date: 2016-10-12
TIANJIN UNIV
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[0003] In order to solve the problems in the prior art, the present invention provides a method for detecting Vibrio parahaemolyticus in coastal seawater, which solves the problem of time-consuming detection of Vibrio parahaemolyticus based on the selective culture method in the prior art, which consumes a lot of manpower and material resources , The problem of being unable to quantify non-culturable Vibrio parahaemolyticus in seawater when detecting Vibrio parahaemolyticus
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[0023] The present invention will be described in detail below in combination with specific embodiments.
[0024] The method established in this study is suitable for the quantification of Vibrio parahaemolyticus in seawater samples. The method description is as follows:
[0025] (1) At the set sampling point, collect a water sample at 0.5m below the water surface with a special sample collector for water samples, put it into a sterile glass bottle with a volume of 1L, and store it in an ice box after numbering. After the sample collection is completed, it is immediately transported back to the laboratory at low temperature for processing. After transporting back to the laboratory, some water samples are filtered with a microporous filter membrane with a pore size of 0.22 μm. Each filter membrane filters 500 mL of water samples. Three filter membranes were filtered at the sampling point, and the filter membranes were stored in a -80°C ultra-low temperature refrigerator for sub...
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The invention discloses a detecting method of vibrio parahemolyticus in coastal seawater. The method includes the steps of extracting DNA of an environment sample, amplifying a target segment, establishing a standard plasmid template, establishing a Q-PCR quantitative standard curve, and measuring the number of vibrio parahemolyticus in the environment sample. Compared with current detection based on a selective culture method, the method has the advantages that detecting time is shortened from 10-12 days of the existing method to 1 day, and only 3-4 hours are needed if the standard curve is completed; furthermore, the problem that the existing method can not measure uncultured vibrio parahemolyticus is solved, and the identification accuracy is improved.
Description
technical field [0001] The invention relates to the technical field of common bacteria detection, in particular to a method for detecting Vibrio parahaemolyticus in coastal seawater. Background technique [0002] Vibrio parahaemolyticus (Vibrio parahaemolyticus) is a Gram-negative halophilic bacterium that mainly exists in coastal seawater, seawater sediments, fish, shrimp, shellfish and other seafood. Human consumption of raw or undercooked seafood contaminated by this bacteria can cause diseases such as acute gastroenteritis, reactive arthritis, and even sepsis in severe cases. Therefore, the detection of Vibrio parahaemolyticus in food has received extensive attention. As a marine environment closely related to human activities, coastal seawater, especially bathing beaches, is affected by Vibrio parahaemolyticus. However, the current domestic detection methods for Vibrio parahaemolyticus in coastal seawater mainly use traditional bacterial isolation and biochemical iden...
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Login to View More IPC IPC(8): C12Q1/68C12Q1/06
CPCC12Q1/6851C12Q2545/114C12Q2531/113Y02A50/30
Inventor 商君阳何艺科汪光义
Owner TIANJIN UNIV