SPR Biosensor Detection For Serum Tryptase In Animal Of Anaphylactic Shock

Research backgroundAnaphylactic shock is an intensly syndrome involving multipleorgans happened in a short period of time by immunologic mechanismwhen some antigenic substance from the outside world attact the sensitivedbody. in severe cases， they will enter a state of shock. if rescue notprompt, the patients will die within5-10min. Anaphylactic shock has beencaused great trouble in the forensic medicine,because it has no specificpathological changes.When the body firstly contacts allergens,it will produce specificIgE. The specific IgE adsorbing on the surface of mast cells would makethe body in a state of sensitization. When the body contacts the allergenagain, the allergen will combine with the specific IgE adsorbing on thesurface of mast cells.It leads to the activation of mast cells and the releaseof bio-active substances.This is the key link of anaphylactic shock.Thebio-active substances include histamine, kininogenase, LTs, PAF and soon, but the half-life of bio-active substances lasts only a few seconds to afew minutes,and these mediums are extremely unstable.after death. Tryptase is a kind of specific protein released in the degranulation of mastcells. Tryptase has a long half-life which is greater than1h,it can keepstable after death. The average content of normal serum tryptase is2.24ng/mL, the serum tryptase mostly cannot be detected in normal people.Studies have shown that:the serum tryptase concentration has a slightlydifference between different countries.It is generally considered thatserum tryptase concentration in normal people cannot surpass5ng/mL,while the serum tryptase concentration in people death from anaphylacticshock can achieve dozens of times of normal people. The existing domesticand foreign scholars have been used immunohistochemical methods、radioimmunity methods and ELISA to detect the body blood content oftryptase to diagnose anaphylactic shock..ObjectiveUsing surface plasma resonance (SPR) technology to detect serumtryptase in guinea pig death from anaphylactic shock, trying to provide abasis for objective diagnosis in death from anaphylactic shock. Analyzingthe the influence to the result of the experiment by using two different SPRanalysis methods.MethodsThe study chooses healthy guinea pigs as animal models. First of all,16healthy guinea pigs are randomly divided into experimental group and control group． For each group animals are sub-grouped intoanimals of instantaneous death and animals kept at-20oCfor three days freezing. Guinea pigs in instantaneous death group ofexperimental group are divided into3h,5h,10h,20h group Anaphylacticshock is induced by injection of human pooled serum.Anti-TPSAB1areimmobilized on the surface of SPR biosensor chips using self-assemblymethod, and then SPR is adopted to detect the tryptase in guinea pigsserum.Using the way of passing into the channel and the way of titrationrespectively to detect serum tryptase in guinea pig in instantaneous deathgroup.ResultsThe response value of experimental group of tryptase is significantlyhigher than the control group. The response value of three daysfreezing group of tryptase has no significant difference with instantaneousdeath group.The three days freezing condition dose not affect theexpression for serum tryptase. The serum tryptase in guinea pig of the3h,5h,10h,20h groups of experimental group has no obvious change.Thedeath after20h condition dose not affect the expression for serum tryptase.explain how about20h does not affect the guinea pig serum tryptaseexpression.Using the way of passing into the channel and the way oftitration respectively to detect serum tryptase in guinea pig cannot change the results.It shows that the two ways can both detect the serum tryptase.ConclusionBiosensor based on SPR has good application prospectin detection of animal serum tryptase，and expression ofdetectio n of tryptase may act as an evidence fordiagnosis of anaphylactic shock．...