Detection Of Aquatic Pathogens By Noveltime-Resolved Fluoroimmunoassay

The scale of aquaculture in China is among the highest in the world.With the development of the high-density intensive aquaculture model,diseases in the aquaculture process occur more and more frequently,among which bacterial diseases are the most serious.The outbreak of bacterial diseases not only restricts the development of aquaculture industry,but also has a potential threat to human health.Therefore,it is very important to establish and perfect the early diagnosis method of aquatic pathogenic bacteria.Time-resolved fluoroimmunoassay?TRFIA?has become one of the most sensitive biological detection techniques with its advantages of high sensitivity,low background,simple to use and no radioactive pollution.It is well worth further researching and developing its potential applications and values.In view of the advantages of TRFIA method,our team first established a set of mature new methods for the detection of aquatic pathogenic bacteria.The purpose of this paper is to explore new research methods on this basis.The main contents are as follows:?1?In this paper,the bifunctional europium chelate ETNP was synthesized and its fluorescence properties were determined.Aeromonas hydrophila B11 antibody was labeled by ETNP in the presence of glutaraldehyde.The direct solid phase TRFIA method was established for the detection of Aeromonas hydrophila B11 with a detection limit of 5×10^-6 cfu/mL.?2?A new bifunctional europium chelate ETBCP was synthesized,and a novel solid phase TRFIA method was established for the detection of Aeromonas hydrophila B11.The detection limit was 1×10⁵ cfu/mL.The new method was used to detect the tissue samples of infected eel,the results were satisfactory and the sensitivity has improved.?3?We have introduced the co-luminescence effect of rare earth elements into DELFIA system,a directly new co-luminescence TRFIA method was established.The sensitivity of the method was greatly improved with the introduction of inert rare earth ion Lu³⁺.The detection limit of Aeromonas hydrophila B11 was 1×10³ cfu/m L,which was two orders of magnitude lower than that of direct detection with purchased Eu³⁺labelling kit.