Method for detecting dot-ELISA of principally outer membrane protein of vibrio vulnficus gene

A technology of the main outer membrane protein and Vibrio vulnificus, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of less quantitative detection research, insufficient attention, insufficient sensitivity of the method, and insufficient consideration of feasibility, etc., to achieve The effect of high sensitivity and specificity, convenient preparation and simple operation
CN101451996AInactive Publication Date: 2009-06-10张振冬
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
张振冬
Publication Date
2009-06-10
Estimated Expiration
Not applicable · inactive patent

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Abstract

Disclosed is dot-ELISA detection method of main outer-membrane protein of vibrio vulnficus. Cultured vibrio vulnficus is crashed (power 200w, beyond 5s stopping 5s) through ultrasonic, supernatant is centrifugally collected. Spotting 1.0 mu l is on a polyvinylidene fluoride film (PVDF film), after the spotting PVDF film is closed by 2% BSA (bovine serum albumin) under 37 DEG C, respectively reacted with rabbit anti vibrio vulnficus serum and goat anti-rabbit IgG whole serum marked by HRP (horseradish peroxide), finally reacted with substrate solution, macroscopic brown yellow round sedimentation is formed on the PVDF film, namely masculine result. The inventive method has detection sensitivity 15 ng / ml for main outer-membrane protein of vibrio vulnficus, is 3-5 times higher than enzyme linked immunosorbent assay (ELISA), the result is obtained after 2-3 hours, therefore vibrio vulnficus in the marine environment is detected fast and sensitively.
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Description

technical field

[0001] The invention relates to a rapid detection method for the main outer membrane protein of Vibrio vulnificus in marine environment. Background technique

[0002] Vibrio vulnificus (Vibrio vulnificus) is a marine bacterium that is widely distributed in seawater and sediments in coastal waters, estuaries and bays. Vibrio vulnificus can cause serious diseases such as human food poisoning and sepsis through intestinal and trauma infection. Shellfish such as oysters are the main medium of Vibrio vulnificus infection, which is mostly caused by eating contaminated shellfish raw. It has been found in coastal areas of my country in recent years The cases of acute diarrhea and sepsis caused by Vibrio vulnificus are gradually increasing. At the same time, Vibrio vulnificus is also the main pathogen that endangers marine shellfish and fish farming in my country, causing serious economic losses to related industries. However, the qualitative and quantitative detectio...

Claims

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