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Method and kit for detecting VBNC vibrio parahemolyticus

A technology of hemolytic vibrio and detection method, applied in the field of PMA-qPCR detection method, can solve the problem of inability to distinguish between active bacteria and inactive bacteria, and achieve the effects of stable results, good repeatability and high sensitivity

Pending Publication Date: 2017-11-17
PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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AI Technical Summary

Problems solved by technology

Although it has the advantages of short time consumption and high accuracy, conventional qPCR can only detect the total number of bacteria in food, and cannot distinguish active bacteria from inactive bacteria

Method used

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  • Method and kit for detecting VBNC vibrio parahemolyticus
  • Method and kit for detecting VBNC vibrio parahemolyticus
  • Method and kit for detecting VBNC vibrio parahemolyticus

Examples

Experimental program
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Embodiment

[0033] 1. Strains

[0034] Vibrio parahaemolyticus (ATCC17802), Vibrio alginolyticus (ATCC 33787), Vibrio vulnificus (ATCC 27562), Enterococcus faecalis (ATCC 49452), Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Escherichia coli (ATCC 35218), and Edwardsiella tarda (ATCC 15947) were purchased from Shanghai Hanni Biotechnology Co., Ltd. Vibrio cholerae (VbO FSCC 232004) was purchased from Guangdong Food Microbial Safety Engineering Technology R&D Center. All strains were stored at -80°C in 10% (w / v) glycerol broth.

[0035] 2. Method

[0036] 1. Resuscitating strains: Inoculate 100 μL of bacterial solution from a glycerol cryopreservation tube into 5 mL of brain heart infusion culture broth, shake overnight at 37°C and 150 r / min.

[0037] 2. Sample pretreatment and VBNC induction in frozen state: sample collection was carried out in accordance with the relevant provisions of GB4789.1-2010 "National Food Safety Standard Food Microbiology Inspection...

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Abstract

The invention relates to a method for detecting VBNC vibrio parahemolyticus in a freezing matrix by means of a PMA-qPCR method and primer and probe combination implementing the method. The PMA-qPCR detection method provided by the invention is high in specificity, high in sensitivity and good in repeatability in vibrio parahemolyticus detection and is suitable for detecting viable but nonculturable vibrio parahemolyticus in a frozen food.

Description

technical field [0001] The invention belongs to the field of food inspection and quarantine and molecular biology detection, and specifically relates to a detection method and a reagent for Vibrio parahaemolyticus, in particular to a PMA-qPCR detection method for VBNC Vibrio parahaemolyticus in a frozen matrix. Background technique [0002] Vibrio parahaemolyticus (V.parahaemolyticus) is a Gram-negative halophilic bacterium that is widely distributed in oceans and salt lakes, and mainly pollutes marine fish, shrimp, shellfish, and shrimp. Cause acute gastroenteritis, clinical symptoms include diarrhea, vomiting, nausea, abdominal cramps, low-grade fever, chills, etc. Nearly 50% of food poisoning is caused by Vibrio parahaemolyticus, which is an important food-borne pathogen . As an important port in the country, Shanghai imports a large amount of seafood every year, especially chilled products, such as Atlantic salmon, tuna, sea urchin, etc. Vibrio parahaemolyticus is a mus...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06C12N15/11
CPCC12Q1/6851C12Q1/689C12Q2563/107C12Q2563/173Y02A50/30
Inventor 黄新新杨娟蔡强
Owner PLANTS & ANIMALS & FOOD TESTING QUARANTINE TECH CENT SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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