Kit and method for detecting vibrio parahaemolyticus in aquatic product

Abstract

The invention discloses a kit and method for detecting the vibrio parahaemolyticus in an aquatic product. The kit comprises sterile deionized water, PCR (Polymerase Chain Reaction) liquid, TaqDNA (Deoxyribonucleic Acid) polymerase, an ethidium bromide DNA dye, a bromophenol blue loading buffer, a standard sample and a control sample. The detection method comprises the following steps: firstly collecting samples, carrying out enrichment culture on the samples, extracting the DNAs after the enrichment culture, preparing PCR reaction tubes, carrying out PCR amplifications on the extracted DNAs on a PCR instrument respectively to obtain PCR amplification products, and judging the detection results. The kit has the following advantages that the kit can be applied to the aspects of the fast detection of the vibrio parahaemolyticus, the epidemiological investigation, the drug efficacy monitoring and the like. Compared with an existing detection method, the method disclosed by the invention is convenient to use, good in reliability, short in detection period, high in sensitivity, strong in specificity, low in cost, simple in operation step and suitable for high throughput operation and standard operation, and is of great significance in improving the safety level of food.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to a kit and method for detecting vibrio parahaemolyticus in aquatic products. Background technique [0002] Vibrio parahaemolyticus ( Vibrio parahaemolyticus ) is a halophilic bacterium belonging to the genus Vibrio in the family Vibrio. Germs. After eating food contaminated by Vibrio parahaemolyticus, it can lead to acute gastroenteritis with nausea, vomiting, and diarrhea as the main symptoms. The feces of severe patients are watery, mixed with mucus or pus and blood, which may be caused by dehydration and shock. coma or even death. Food poisoning caused by Vibrio parahaemolyticus has become one of the foodborne public health problems worldwide. According to literature reports, in Japan, food poisoning caused by this bacteria accounts for 20% to 30% of the total number of food poisoning in the country. Among the bacterial food poisoning in Europe and the United States, t...

AI Technical Summary

Problems solved by technology

At present, the national standard detection method of Vibrio parahaemolyticus (GB / T 4789.7-2008) is the culture method. After the culture method collects the sample, first enrich the bacteria and culture for 24 hours, conduct a preliminary identification according to the bacterial colony morphology, and then confirm it according to the biochemical identification; Its disadvantages are: long culture period, time-consuming and labor-intensive, and low sensitivity to bacteria in the "viable but non-culturable state" (Viable But Non-Culturable, VBNC) that is still pathogenic, which can lead to false negative test results , especially the false negative rate of frozen food is very high, causing great food safety hazards

The reagents and instruments used in the real-time PCR method are relatively expensive, and the requirements for operators are very high; the LAMP method is highly sensitive, but it is very easy to cause aerosol pollution, produce false positives, and cause wrong test results ; ELISA immunological method is complicated to operate, time-consuming and laborious, and not convenient for standardization and high-throughput operation

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